IJLSCI Volume 5 3 September 2017

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28 ISSN:2320-7817 | eISSN:2320-964X
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ISSN: 2322-0015 Cell : (+91) 9970559438 | 9420775527
IJLSCI ISSN:2320-7817(p) |2320-964X(o)
Volume 5, Issue 3, September 2017
Bacopa monnieri (L.) Wettst.,

Family- Scrophulariaceae
Bacopa monnieri (L.) Wettst. locally known as water hyssop,

brahmi or Jalanimba in India. The name brahmi is derived
from the word 'Brahma', the mythical 'creator' in the Hindu
pantheon. As the brain is the center for creative activity, any
compound that improves the brain health is called brahmi,
which also means 'bringing knowledge of the supreme
Contents
362 Community structure and monthly dynamics of
REVIEW ARTICLE
zooplankton in high altitude rice fish system in
Eastern Himalayan region of India
291 An Ethological and Ecological Review of the Saikia Rajashree, Das Tapati, Gogoi Budhin,
real-world applications of H. illucens Kachari Akash, Safi Vivekanand and
Sammer Marzouk Das Debangshu N
ORIGINAL ARTICLE
379 Anatomical markers and Phytochemical study of
different plant parts of Bacopa monnieri (L.)
297 Feeding ecology of tiger fish, (Hydrocynus Wettst.
vittatus) in the Kalimbeza Channel on the
Varshney Anju, Chandra Naresh and
Zambezi River, Namibia
Patil Avinash
Simasiku EK MafwilaSK, Abah J, Hay CJ,
Matengu KK Lichens: A myriad hue of Bioresources with
387
medicinal properties
306 A Morphological Study of the Evolution of Eight Ramya K and Thirunalasundari Thiyagarajan
Organisms Through a Morphological
Phylogeny
394 Antifungal activity of extracts of weed biomass
Sammer Marzouk against Aspergillus flavus Link ex Fr., a causal
agent of yellow mold of Groundnut
311 Effect of integrated use of lime, blended Deshmukh RS
fertilizer and compost on productivity, nutrient
removal and economics of barley (Hordeum Incidence of mycotoxins in poultry feeds and
399
vulgare L.) on acid soils of high lands in West feed ingredients used in Warangal (TS), India
Showa Zone of Ethiopia
Parvathi D, Rajender Reddy A and
Woubshet Demissie, Selamyihun Kidanu and
Krishna Reddy V
Cherukuri V Raghavaiah
405 Phytochemical Screening and Antibacterial
323 Seasonal distribution and abundance of Thrips Activity of Bryophytes
(Thysanoptera: Thripidae) on onion producion
Kadam Pratima S
in central zone of Tigray, Ethiopia
Gebretsadkan Zereabruk
409 Diversity, Distribution and Status of the
Amphibian fauna of Sangli district,
332 The Genetic and Morphological Evolution of Maharashtra, India
Fish through the Protein Cytochrome C
Sajjan MB, Jadhav BV and Patil RN
Oxidase Subunit One
Sammer Marzouk 420 Comparative Study of Antimicrobial Potentials
of Phospholipid Compound Produced by
345 Performance of bread wheat (Triticum Halophilic and Alkaliphiles Bacillus subtilis
aestivum L.) in response to supplemental isolated from Alkaline Meteorite Crater
irrigation and rate of nitrogen application in Lonar lake, India
Enderta, Tigray, Northern Ethiopia
Shinde VA, Patil RB and Pawar PV
Beyenesh Zemichael and Nigussie Dechassa
| IJLSCI Volume 5 | Issue 3|September 2017 | page nos.291- 479 http://www.ijlsci.in

IJLSCI ISSN:2320-7817(p) |2320-964X(o)
Volume 5, Issue 3, September 2017
425 Comparative study to evaluate in vivo and in LIST OF REFEREES

vitro antioxidant potential of processed and
unprocessed Curcuma logna L. using Allxon INTERNATIONAL
induced diabetic rats
Parashare VM, Nikam S Nade VS, Pande Dr. Sushrut Arora, Ph.D.
Shishir Dr. Bela Volgyi, Ph.D.
Dr. Fazal Shirazi, Ph.D.
438 Studies on Physico-chemical analysis of Dr. Bankefa Emmanuel Olufemi, Ph.D.
Mohabala Lake near Bhadrawati, District - Dr. Gurpreet Singh Dhillon, Ph.D.
Chandrapur (MS), India Dr. Mahdi Esmaeilzadeh, Ph.D.
Mahajan VSand Pokale SS Dr. Zohair I.F. Rahemp, Ph.D.
Dr. Hamid Saremi, Ph.D.
447 Effect of weed green manure, compost
manure and vermicompost on productivity of NATIONAL
Spinach
Parbhankar RL and Mogle UP Dr. Jaykiran A. Tidke, M.Sc. Ph.D.
Dr. Surendra R. Manik, M.Sc. Ph.D.
451 Physico-Chemical parameters of Nizam sagar
Dr. P. A. Wadegaonkar, M.Sc. Ph.D.
dam with respect to fish production
Dr. D. H. Tambekar, M.Sc, PhD
Phytoplankton, Zooplankton
Dr. R. N. GACCHE, M.Sc. Ph.D.
Totawar DV and Tamlurkar HL
Dr. B.N. Pandey, M.Sc. Ph.D.
455 Dr. Satish A. Bhalerao M.Sc. Ph.D.
A new species of cestode Lytocestus
Dr. Kamble SY, M.Sc. Ph.D.
paithanensis (Lytocestus Cohn, 1908) from
Dr. Suresh B. Zade, M.Sc. Ph.D,
Clarias batrachus at Paithan, MS, India
Dr. Varsha Wankhade M.Sc. Ph.D,
Kale SS
Dr. Gunjan Mukherjee, Ph.D.
459 Dr. Santosh Pawar, M.Sc. Ph.D.
Impact of temperature variations on RNA
Dr. Rajusing G. Jadhao, M.Sc. Ph.D
content of Freshwater fish Oreochromis
mossambicus Dr. Rathod Mulchand M. M.Sc. Ph.D
Chavan PN and Mali RP Dr. Bhajbhuje M.N. M.Sc. Ph.D.
Dr. Kishor G. Patil, M.Sc. Ph.D.
462 Systematic study of Cestode parasite Dr. Mali Rajendra P. M.Sc. Ph.D.
Moniezia (Blanchariezia) Caprae n. sp. of Dr. D.U. Gawai M.Sc. Ph.D.
Capra hircus from Kasgi Tq. Omerga Dist. Dr. Dhore Mukund M., M.Sc. Ph.D. D.
Osmanabad, (MS) India Dr. Gyan Singh Shekhawat M.Sc. Ph.D.
Kale Sanjay S Dr. K. Madhava Chetty, M.Sc. Ph.D.
Dr. Mumtaz Baig. Ph.D. D.Sc.
465 Sarus Crane in Indian Mythology and Culture Dr. Sushant Punekar M.Sc. Ph.D.
Kumar Adesh and Kanaujia Amita Dr. Varsha Zade., M.Sc. Ph.D.
Dr. Dinesh Khedkar, M.Sc. Ph.D.
471 Burud Society and Bamboo Plants Utility : Dr. Dilip Hande, M.Sc. Ph.D.
Dendrocalamus strictus Dr. Bhalerao S.S., M.Sc. Ph.D.
Deshmukh Pratap V Dr. Dimdime D.S., M.Sc. Ph.D.
Dr. Vishal Marathe M.Sc. Ph.D.
Dr. Deosthale D.V. M.Sc. Ph.D.
OVERVIEW Dr. Rajendra V. Tijare, M.Sc. Ph.D.
Dr. Sangita P. Ingole M.Sc. Ph.D.
474 Assessing the impact of pesticides: An Dr. S.S. Nanware M.Sc. Ph.D.
overview Dr. (Mrs.) Gayathri Narayan M.Sc. Ph.D.
Jawale CA, Rajput KH and Ugale BJ Dr. Patil S.D. M.Sc. Ph.D.
Dr. Deshmukh CK, M.Sc. Ph.D.
480 Author index Dr. Deshmukh.Sanyogita, M.Sc. Ph.D.
Dr. Kumud Choudhary M.Sc. Ph.D.
481 Author Instructions Dr. Piyush Gupta M.Sc. Ph.D.
Dr. Tushar Wankhede M.Sc. Ph.D.
Dr. Sachin Tawade M.Sc. Ph.D.
Dr. Ade Suraj., M.Sc. Ph.D.
Dr. Aruna U. Kakade M.Sc. Ph.D.
Dr. Jayvardhan V. Balkhande M.Sc. Ph.D.
,http://www.ijlsci.in Volume 5 | Issue 3|September 2017 | page nos. 291- 480 | IJLSCI
Int. J. of Life Sciences, 2017, Vol. 5 (3): 291-296 ISSN: 2320-7817| eISSN: 2320-964X
REVIEW ARTICLE
An Ethological and Ecological Review of the real-world

applications of H. illucens
Sammer Marzouk
University of Chicago Laboratory Schools, 1362 E 59th St, Chicago, IL 60637,

Email: marzouk.sammer@gmail.com
Manuscript details: ABSTRACT

Received: 19.07.2017 Introduction: The Black Soldier Fly (Hermetia illucens) is a true fly in the
Accepted: 13.08.2017 family Stratiomyidae and subclass Pterygota. H. illucens has been a topic of
Published : 09.09.2017 interest in the ecological research world for many years. H. illucens
possesses special physiological, ethological and ecological features that may
Editor:
allow H. illucens to become a model organism.
Dr. Arvind Chavhan
Cite this article as: Results: This literature review looks at the physiological, ethological and
Sammer Marzouk (2017) An ecological features of H. illucens. Existing literature about H. illucens has
Ethological and Ecological Review of been searched and analyzed for this paper. From the literature search, it
the real-world applications of was discovered that H. illucens is not known to have valuable impacts to the
H.illucens; International J. of Life world at large. The purpose of doing this review is so that all information
Sciences, 5 (3): 291-296. available on H. illucens can be chronicled and so the information can be used
in real-world applications.
Copyright: 2017| Author (s), This Conclusions: It provides a use of H. illucens as a model organism and aims
is an open access article under the
to demonstrate a framework for real-world applications of H. illucens, such
terms of the Creative Commons
as H. illucens becoming a model organism for mosquitoes, Tsetse flies, and
Attribution-Non-Commercial - No
Derivs License, which permits use house flies.
and distribution in any medium, Keywords: Individual Ecology, Population Ecology, Behavioral Ecology,
provided the original work is
Mating, Ethology, Ecology.
properly cited, the use is non-
commercial and no modifications or
adaptations are made.
INTRODUCTION
There are more than 1,000,000 species of Diptera on planet Earth. Most of
these species of Diptera are organisms that require little care, take up little
space and can be efficiently be raised. The purpose of this literature review
is to gather all the available information on H. illucens that is related to
raising H. illucens from larvae into adulthood and gathering the most
amount of offspring from the adults. Existing literature about H. illucens has
been searched and analyzed for this paper. From this information, the idea
of H. illucens being used as a model organism is presented. Using H. illucens
as a model organism allows the advantage of an easy to take care of, easy to
grow organism with evolutionary and biological connections to organisms
that present medical and financial problems to specific groups of people.
2017 |IJLSCI www.ijlsci.in 291

Sammer Marzouk, 2017
MATERIALS & METHODS lot as larvae as when they are adults they cannot eat,
as they lose their mouthparts (Rozkosny, 1983). The
Obtaining Research Papers energy they gain from being larvae is the energy they
For the literature review, the research papers were have for the rest of their lives (Rozkosn, 1983). H.
obtained from Web of Science. Web of Science is an illucens as a pupa is dark brown in colour (Rozkosn,
online database that allows access to thousands of 1983). As adults, H. illucens are black. (Rozkosn,
papers. The papers were all allocated to the following 1983). Because of its white tarsi, black legs, and
topics: H. illucens, behavior, sexual dimorphism and elongated antenna H. illucens is a Batesian mimic of the
ecology. These papers were all copyrighted to the wasp (Rozkosn, 1983). They also have two
public. There were 26 papers on H. illucens and related transparent on their abdomen, allowing them to have a
topics that were chosen for the literature review. slender, wasp-like abdomen (Rozkosn, 1983). They
also have changed their behavior so that they act like
Procedure wasps. This includes the way they stand, move and
For the procedure, the 26 literature reviews were read flight patterns (Rozkosn, 1983). Adults range from 15
with the intent and purpose of specifically finding the to 20 mm in length (Sheppard et al. 2002). They live
part of the papers that went along with the for about 5-8 days as adults (Newton, 2005).
information that was to be presented on H. illucens.
After the specific information was gathered, if was Ethology:
organized into the topics of H. illucens which includes As larva, the flies have only one purpose and that is to
behavior, sexual dimorphism, and ecology. After the eat (Tomberlin et al., 2013). The reason why is
information was organized, the information was because the energy that they get as a larva is the
expanded into short paragraph form, which was then energy that they will use for the rest of its life
combined into the final draft of the paper. (Tomberlin et al., 2002). They move to get their food
and only eat as that is their only goal as a larva. After
they are ready to pupate, the larvae then move to a
RESULTS new area, preferably dry and cold to go through the
transformations (Newton, 2005) When an adult H.
Anatomy/Physiology: illucens is territorial, as they follow a lekking system
H. illucens has various sexually dimorphic traits such (Tomberlin et al. 2001). This is because when it is time
as body size, features on the head, abdominal spots to mate, all the males try to find spots in the lekking
and the shape of their abdominal terminalia to area (Tomberlin et al. 2001). The females only mate
differentiate sex. Females have more whitish hairs on with the males who have territory to mate on
their face. Females only have four black frontal (Tomberlin et al. 2001). Males that do not have any
tubercles, and female abdomens are also longer territory do not have anywhere to mate (Tomberlin et
(Rozkosn, 1983). The cerci have two segments al. 2001). The more territory a male has, the greater of
(Rozkosn, 1983). The females are also smaller than a chance they have or mating. If another male comes in
the males (Rozkosn, 1983). The males have fewer the area, the two will fight, and the winner will take
whitish hairs on their face. (Rozkosn, 1983). For the territory (Newton, 2005). The most popular
males, the genitalia is relatively shorter than the territories are the ones that have had the most mating
female terminalia (Rozkosn, 1983). The genitalia is in (Tomberlin et al. 2001). H. illucens are not extremely
alignment with the syn sternum, possessing two competitive. Even if another male comes on their land,
posterolateral lobes on each side (Rozkosn, 1983). As they are not going to immediately fight them, only if a
adults, female wing sizes are bigger than the males. female is coming will the two engage (Tomberlin et al.
Males had an average wing size of 14.24 mm, which 2001). Body size does not matter when it comes to
females had an average wing size of 14.67mm (Gobbi mating as females mate with the first male that they
et al., 2013). As larvae, H. illucens can reach 27 mm in see, regardless of size, (Tomberlin et al. 2001). Also
length and six mm in width (Rozkosn, 1983). They because H. illucens are not competitive, a smaller male
are a dull, whitish color with a small, projecting head can come onto lekking territory, and mate with a
containing chewing mouthparts (Rozkosn, 1983). female on another male's territory (Tomberlin et al.
Larvae pass through six instars and require 14 days to 2001). Direct sunlight encourages mating for H.
complete development (Rozkosn, 1983). They eat a illucens, and it has rarely been recorded mating on
292 Int. J. of Life Sciences, Vol. 5(3) September, 2017

An Ethological and Ecological Review of the real-world applications of H.illucens
rainy or snowy days (Zhang et al., 2010). Recorded in compositions including animal waste, spoiled fruits
North America, mating activity begins at 8:00 am, and and vegetables, fish rendering waste, and carrion
peaks at 10:00am CDT (Zhang et al., 2010). (Tomberlin et al., 2013). These habitats can help, or
Oviposition occurs on the 17th day, and has occurred hinder larval development (Tomberlin et al., 2013).
without sunlight, but had a low chance (Tomberlin et This is because of the fish waste and animal waste
al. 2001). Females also had mating issues as females hinder larval development, and leads to higher
with larger wing sizes and larger bodies are more mortality rates (Tomberlin et al., 2013). Fruits and
fertile than those with smaller bodies and smaller vegetables decrease mortality, speed up larval
wings (Gobbi et al., 2013). After mating, females lay development, and increase overall body size for H.
their eggs on cracks and crevices near decaying matter illucens as adults (Tomberlin et al., 2013). The main
(Zhang et al., 2010). As larvae, H. illucens feed on the predators of H. illucens as both larvae and adults are
decaying organic material, they were born next to other Dipterans (Peterson et al., 1960). These include
(Dicarlo et al. 2010). But as adults, the flies cannot eat, Empididae, Dolichopodidae, Ephydridae, and
they can only drink water (Tomberlin et al., 2013). Tendipedidae (Peterson et al., 1960). Predators in
This is because their mouth cannot chew (Tomberlin other genuses include Trichoptera, especially species
et al., 2013). Towards the end of being a larva, H. of the genus Hydropsyche and Odonata (Peterson et
illucens becomes a puparium (Tomberlin et al., 2013). al., 1960). Hymenoptera is a parasitoid that only
A puparium is the last larval stage before pupating affects eggs of H. illucens (Peterson et al., 1960). Non-
(Tomberlin et al., 2013). As a puparium, they turn insect predators include spiders and leeches which
black from releasing melanin, a chemical into their only eat the larvae (Peterson et al., 1960). Frogs are
skin that increases the durability of the skin also well-known predators of adult males because
(Tomberlin et al., 2013). They also enter a wandering males go near bodies of water during mating season
stage and start to wander (Tomberlin et al., 2013). (Peterson et al., 1960). H. illucens larvae are
They wander because they are looking for a place to detritivores and have no real prey (Peterson et al.,
pupate (Tomberlin et al., 2013). This is a preferably a 1960). Light affects mating because direct sunlight
dry and cold place like an underground tunnel, and encourages mating for H. illucens (Zhang et al., 2010).
they also stop eating (Tomberlin et al., 2013). When H. illucens has rarely been recorded mating on rainy or
they find a proper habitat, they stop moving and snowy days because the light level is too low (Zhang et
connect to the ground vertically. (Tomberlin et al., al., 2010). In a laboratory setting, the level of light with
2013). the most amount of mating is 200 mols / m^2.
Insects cannot see infrared or red light (Zhang et al.,
Ecology 2010). The shortest wavelength they can see up to is
H. illucens has a range from the Americas to South-Asia 700 nm (Zhang et al., 2010). Members of Diptera only
(Rozkosn, 1983). They live in areas with lots of have light receptors for high blue or low green colors
decaying matter (Rozkosn, 1983). At all stages, H. (480-530 nm) (Zhang et al., 2010). Adult size is
illucens has a narrow range of temperatures they can determined by pupal development (Gobbi et al., 2013).
live in, between 27-30 degrees Celsius (Rozkosn, The more energy the larvae was able to gather, the
1983). At 36 degrees or higher, they die off. H. illucens bigger the adult will be (Gobbi et al., 2013). The
is not active in winter months, due to the temperature, amount of food that it receives as a larvae limits its
and because the adults die after breeding, leaving the growth. The diet also affects the mass of the flies,
eggs that will hatch in spring (Tomberlin et al., 2013). which affects fecundity (Gobbi et al., 2013). Females
Relative humidities of 3090% were enough for with more mass had more offspring than those with a
mating, but the higher the humidity, the larger the smaller amount of mass (Gobbi et al., 2013) The
larvae is. (Holmes et al. 2012). As larvae, the H. illucens temperature and gender of H. illucens can factor
eat decaying organic material, like food scares and growth and fitness. At 27 and 30C, females weighed an
kitchen waste (Nguyen et al. 2012). But as adults, the average of 17-19% more than males (Tomberlin et al.,
flies only drink water (Nguyen et al. 2012). In order for 2009). But males lived 20% longer (Tomberlin et al.,
H. illucens to achieve mating reliability, they must be in 2009). H. illucens are r selected (Tomberlin et al.,
a space at least 0.76 by 1.14 by 1.37m or 10 liters 2009). This is because they have about 320-620 eggs
(Sheppard et al., 2012). The H. illucens habitat is in a big bang reproduction and die off hours later
patchily distributed and includes various organic (Tomberlin et al., 2009). The eggs then grow up with
www.ijlsci.in Int. J. of Life Sciences, Vol. 5(3) September 2017 293

no parental help (Tomberlin et al., 2009). After the (Brown et al., 1997). These insects put their offspring
eggs are laid, it takes about 3-4 days for the eggs to in the eggs of Dipterans, and take the nutrients away
hatch (Furman et al., 1959). They stay as larvae for until the eggs die. After that, the Hymenoptera hatches
two weeks, and in the pupal stage for two weeks, but and leaves (Brown et al., 1997).
this period can increase to four months if resources
are limited (Furman et al., 1959). As adults, the flies
live for 5-8 days. H. illucens live in random DISCUSSION
distributions (Tomberlin et al., 2002). For larva, this is
determined by the amount of supplies, and how they H. illucens and the knowledge that can be derived from
are spaced. In the pupal stage, this is determined by it can be used beyond the implications of just research,
finding an adequate habitat to pupate (Tomberlin et to field such as forensics, commercial and medical use,
al., 2002). For adults, this is determined by gender, to even recreational gardening. In the field of forensics,
size, and territory (Tomberlin et al., 2002). This is H. illucens could be used to find the Minimum
because males and females live in different areas Postmortem Interval (PMI) of a dead body (Nguyen et
before mating, and those who are larger live in better al. 2013). Since H. illucens have their larvae in places
areas than those who are smaller (Tomberlin et al., with dead organic matter like a human, it could be
2002). If there is better territory in a close radius, than used with the knowledge of the growth of H. illucens
they flies will be close (Tomberlin et al., 2002). If it is larvae in specific temperatures and with specific
spread out, then the flies will be spread out resources to find the date of death. For example, given
(Tomberlin et al., 2002). In competition, H. illucens as a person who was killed on a 26-degree day and they
an adult does not have any interspecific competition as have fruits and vegetables in their stomach, the
they only drink water (Tomberlin et al., 2002). They information gathered about larval development could
have intraspecific competition with each other for be used to find when the soldier flies deposit their
mating (Tomberlin et al., 2002). Larvae also do not larvae into the body (Nguyen et al. 2013). This time
have much interspecific competition (Tomberlin et al., can be used to give a date of when that person was
2002). This is because the larvae are detritivores and killed and can be used as evidence in a jury to prove or
do not have competition for dead organic matter in the disprove innocence (Nguyen et al. 2013). H. illucens
places that they live in (Tomberlin et al., 2002). The can also be used to gain cheaper feed for farm animals
larvae may compete with themselves in intraspecific (Tomberlin et al., 2002). They eat a lot as larvae, and
competition in there are limited resources (Tomberlin when pupating releases, a chemical called melanin
et al., 2002). And sometimes, other fly larvae may which turns the pupal black (Tomberlin et al., 2002).
compete with H. illucens larvae for a resource After it pupates, the vitamin count of a pupal is about
(Tomberlin et al., 2002). H. illucens is in a mutualistic that of a soybean, but cheaper and grows at a faster
relationship with some species of fungi (Brown et al., rate (Tomberlin et al., 2002). And it is edible by almost
1997). This is because H. illucens put their eggs on every farm animal (Tomberlin et al., 2002). H. illucens
dead fungi, giving them a food source (Brown et al., as a food can be grown directly at the farm (Newton,
1997). When the eggs grow into larvae, they eat the 2005) This is because as it eats decomposing organic
dead fungi, and other species of plants in the area matter, like animal feces (Newton, 2005). This could
(Brown et al., 1997). This gives the fungi more room to save farms money as previously, they would have to
grow. And when the adults die, they make the soil import their food from other locations, wasting gas
more nutritious for the fungus (Brown et al., 1997). H. and transportation money (Newton, 2005). It is
illucens plays a pivotal role in communities, as they estimated that H. illucens can save a company $25,000
regulate waste disposal (Brown et al., 1997). H. illucens per year per cage of livestock (Newton et al., 2005). H.
helps to keep ecosystems in check because they can illucens as larvae is used as food for fish camps, in
keep the nitrogen and phosphorus levels in an order to make fish bigger (Kroeckel et al., 2012). This
ecosystem stable by eating unwanted manure that is because as larvae, they have a large amount of
could damage the cycles (Furman et al., 1959). Also, H. calcium which is important for fish development
illucens keeps pest flies away because they eat the (Kroeckel et al., 2012). This is not often found in a
rotting organic matter, leaving the pest flies without a fish's diet and helps fish become bigger (Kroeckel et
food source (Furman et al., 1959). H. illucens also al., 2012). This brings the advantage of H. illucens
affected by parasitoids, especially Hymenoptera larvae decreasing the amount of pollution from

An Ethological and Ecological Review of the real-world applications of H.illucens
manure (Newton, 2005). A problem of this generation the sound of the wings flapping (Aksoy et al., 2014). If
is that the phosphorus and nitrogen cycle have been the females flap their wings louder, they get a mate
corrupted by the use of artificial means of gaining faster. From H. illucens, weaknesses such as the cold
these chemicals animals, especially cows. Companies weather and having to hear sounds to mate could be
make too much manure, and allow it to pile up because used in order to decrease mosquito populations
they cannot use it for anything else. This allows it to (Aksoy et al., 2014). The could contribute in lowering
leak into water sources, which contaminates drinking Malaria rates and stopping Mosquito related diseases
water. H. illucens as larvae are unsatisfiable eaters, and across the world. Houseflies can spread typhoid fever
if put into manure can decrease the amount by up to and yaws (Aksoy et al., 2014). But like H. illucens, they
42-64% (Newton, 2005). From the remaining compost, both get advantages from humidity (Aksoy et al.,
the adults would die on it after giving birth, making it 2014). The higher the humidity, the faster the eggs will
of a higher quality (Newton, 2005). H. illucens has also grow (Holmes et al. 2012). So low humidity areas
been used in the production of sugar and biodiesel make it harder for the fly eggs to hatch (Holmes et al.
from dairy manure (Li et al. 2011). The use of H. 2012). Both H. illucens and house flies also have to put
illucens in farms can make food production safer their eggs in organic matter so that they can get energy
(Newton, 2005). H. illucens, when placed in manure as they get older (Aksoy et al., 2014). If mating areas
and chicken coups lowered the amount of diseases, are removed, then the amount of house flies in an area
most notably E.Coli and Salmonella in the food will decrease (Aksoy et al., 2014). This would help to
(Erickson et al. 2014).This is because the larvae alter increase food and water safety, as well as stopping
the chemical composition in mature, making it harder avoidable deaths. The Tsetse fly spreads Sleeping
for the diseases to form (Erickson et al. 2014). This use Disease and has 70 million people in Eastern Africa at
of H. illucens is a cheap and accessible method that risk (Aksoy et al., 2014). Studying the fly is difficult
could be used to increase food safety, and save because the females only lay one egg at a time at
companies money from lawsuits (Erickson et al. 2014). hundreds of locations (Aksoy et al., 2014). The
In the public world, H. illucens has more use than in the similarity with H. illucens and the Tsetse fly is that it
field of food and diseases (Newton, 2005). It can be has been shown that flies share the same genes that
used to discourage other flies from coming to a certain control sex development (Aksoy et al., 2014). This
area (Newton, 2005). H. illucens can be used to get rid means that if the gene in H. illucens were controlled,
of flies because as larvae it is a ravenous eater and eats the gender of the offspring could also be controlled
fast (Newton, 2005). This leaves little food for other (Aksoy et al., 2014). This could allow us to release a
flies (Newton, 2005). Also, the way that it eats, churns strain of this onto a population, making all offspring
its food and leaves it as a liquid (Newton, 2005). This males so that they cannot mate, eliminating a
is a state that the other flies cannot eat as larva, which population (Aksoy et al., 2014)
kills the population over time (Newton, 2005). H.
illucens can also be used in personal gardens (Newton,
2005). They help get rid of pesky insects and diseases CONCLUSION
in the area. Also, they are great at getting rid of
unneeded compost, and make the remaining compost From this review, three conclusions were reached.
better (Newton, 2005). H. illucens also has the They are that H. illucens, through its special
potential of becoming a model organism for physiological, ethological and ecological features can
mosquitoes, houseflies, and Tsetse flies (Aksoy et al., be used as a model organism. H. illucens larvae can be
2014). Tsetse flies spread Malaria and West Nile used in a lab setting because they are easy to take care
Disease by sucking the blood of people (Aksoy et al., of as they will eat decomposing matter (Tomberlin et
2014). H. illucens have to get sexual dimorphism that al., 2002). Larvae can also be put into cramped stations
differentiates males and females (Rozkosn, 1983). in which hundreds of larvae can be stored (Tomberlin
Mosquitos also have sexual dimorphism. Only female et al., 2002). H. illucens adults will need more space to
mosquitoes suck blood, the males do not (Aksoy et al., live, but will still be easy to take care of as the males no
2014). This limits our focus of this problem. The next longer need to eat (Tomberlin et al., 2002). The second
thing is that like H. illucens, mosquitoes are inactive in conclusion of the paper is that H. illucens can be used
winter months due to cold temperatures (Aksoy et al., in an industrial setting in order to cut costs and lower
2014). And finally, Mosquitos locate mates because of multiple types of pollutions on farms. As larvae, H.

illucens eats the decomposing matter of the farm Gobbi P, Martinez-Sanchez A, Rojo S (2013) The effects of
animals and after the larvae become pupae, the pupils larval diet on adult life-history traits of the black soldier
fly, Hermetia illucens (Diptera: Stratiomyidae).
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2017| Published by IJLSCI

RESEARCH ARTICLE
Feeding ecology of tiger fish, (Hydrocynus vittatus) in the

Kalimbeza Channel on the Zambezi River, Namibia
Simasiku EK1*, Mafwila SK2, Abah J3 , Hay CJ 4, Matengu KK5
1Department of Wildlife Management and Ecotourism, Katima Mulilo Campus, University of Namibia, Private Bag
1096, Ngweze, Katima Mulilo, Namibia.
2Department of Fisheries and Aquatic Sciences, and Sam Nujoma Marine and Coastal Resources Research Centre,
Sam Nujoma Campus, University of Namibia, P.O. Box 462, Henties Bay, Namibia.
3Department of Mathematics, Science and Sport Education, University of Namibia, Katima Mulilo Campus Private
Bag 1096, Katima Mulilo, Namibia

4Department of Biological Science, Main Campus, University of Namibia, Windhoek, Namibia.
5Research, Innovation, and Development, Main Campus, University of Namibia, Windhoek, Namibia.
*Corresponding author, e-mail: simasiku2000@yahoo.co.uk
Received: 25.07.2017 The feeding ecology of tiger fish, Hydrocynus vittatus (Characidae), in the
Accepted: 20.08.2017 Kalimbeza Channel on the Zambezi River, Namibia was investigated
Published : 09.09.2017 between February - December 2016. The fish sampling was conducted
monthly using cotton multifilament nets with stretched mesh sizes: 12, 16,
Editor: 22, 28, 35, 45, 57, 73, 93, 118 and 150 mm. A total of 498 specimens of the
Dr. Arvind Chavhan fish were collected and their stomachs dissected for contents analyses. The
findings indicated that large size classes of H. vittatus (>176mm) were
Cite this article as:
largely piscivorous, and showed a diet shift with changes in size. The small
Simasiku EK, Mafwila SK, Abah J ,
size classes of H. vittatus (<140mm) consumed mainly Aquatic insects
Hay CJ , Matengu KK (2017) Feeding
ecology of tiger fish, (Hydrocynus (21.1%), Synodontis spp (17.8%), and Micralestes acutidens (12.1%). They
vittatus) in the Kalimbeza Channel on later shifted to diet in which Synodontis spp (26.1%), Brycinus lateralis
the Zambezi River, Namibia; (15.2%) and M. acutidens (13.0%) dominated. Seasonal shift in diet were
International J. of Life Sciences, 5 (3): apparent due to changes in seasonal food abundances and distributions.
297-305. During the wet season, H. vittatus fed mainly on aquatic insects and later
showed a shift to diet dominated by fish prey items during the dry season.
Hydrocynus vittatus showed no strict food regime and this gives it a better
Copyright: 2017| Author (s), This
chance of survival. Ratios of prey length-predator length averaged
approximately 0.27. The study showed that H. vittatus plays an ecological
Attribution-Non-Commercial - No role with the ability of converting un-exploitable non-commercial species
Derivs License, which permits use into exploitable protein. Hence their population must be of conservation
and distribution in any medium, priority to ensure a balanced fishery in the Zambezi River.
properly cited, the use is non- Keywords: Hydrocynus vittatus, piscivory, predation, Zambezi River

Simasiku et al., 2017
INTRODUCTION Zambezi River, highlighting the effects of fish size and

season on feeding.
The Tiger fish Hydrocynus vittatus (Castelnau, 1861) is
a member of the family Alestiidae, a piscivorous and
ferocious pelagic predator, widely distributed in the MATERIALS AND METHODS
Zambezi River (Jackson, 1961; Skelton, 2001). Larger
specimens up to 700 mm in size, weighing over 150 kg Study area
had been documented (Dalu et al., 2012). This species The Zambezi region borders on Botswana in the south,
are generalist piscivorous (Jackson, 1961; Winemiller Angola and Zambia in the north and Zimbabwe in the
and Kelso-Winemiller, 1993; Mhlanga, 2003; Dalu et east Figure 1a). The area is home to two perennial
al., 2012) and supports an important commercial and rivers, namely the Kwando/Linyanti in the west and
recreational fishery on the Zambezi, Okavango/Chobe the Zambezi Chobe in the east. The Zambezi River is
Rivers and Lake Kariba (Winemiller and Kelso- characterised by a diverse range of habitats including
Winemiller, 1993; Hay et al., 2002). Hydrocynus a deep, wide mainstream with many small vegetated
vittatus is ranked the sixth most important species in islands and sandbanks, small side streams,
the fishery of the Zambezi-Chobe Rivers, contributing backwaters, lagoons and floodplains (Hay et al., 2002).
over 26% to the total annual catch (Hay et al., 2002). The Zambezi floods seasonally, water level starts to
Stomach content studies on freshwater fishes had rise in December with a drastic increase in February
been conducted to provide wealthy of information on and March (Hay et al., 2002).
their dietary requirements in their natural habitats
(Sandon and Tayib, 1953; Verbeke, 1959; Corbet, Field sampling
1961, Chilvers and Gee, 1974; Lewis, 1974 and Whyte, Monthly sampling was conducted in the Kalimbeza
1975). Swallowing the prey whole appears to be a Channel of the Zambezi River (Figure 1b) from
common habit for most freshwater predatory fishes in February to December 2016. The sampling program
Africa (Jackson, 1961). This presents a window that was conducted using a fleet of cotton multifilament
the prey in the gut contents can be observable to nets with stretched meshes of 12, 16, 22, 28, 35, 45, 57,
species level and that both body length and weight can 73, 93, 118 and 150 mm. Freshly caught specimens of
be determined. While several studies have been Hydrocynus vittatus were collected and measured to
conducted on the feeding ecology of H. vittatus (Lewis, the nearest millimetre fork length (SL), weighed to the
1974; Kelley, 1968; Winemiller and Kelso-Winemiller, nearest gram. Fish stomachs were dissected out and
1993; Mhlanga, 2003; Dalu et al., 2012), no study has carefully split open using a pointed nose pair of
systematically analysed their feeding ecology in scissors. Stomach contents were spread out on white
Kalimbeza Channel on the Zambezi River. Kalimbeza treys in the field and visually assessed. Each item in
channel is a newly established fishery reserve on the the diet was identified to the lowest possible
Zambezi River, with minimum exposure to human taxonomic level (using the guides by Skelton, 2001 on
disturbance. Studies on fish feeding can yield relevant fish, Garber and Gabriel, 2002 on insects) counted and
information that can affect competition, mortality, measured to the nearest millimetre total length (TL),
fecundity and growth in fish communities (Michelsen and weighed to the nearest gram.
et al., 1994). Garrison and Link, 2000 highlight the
importance of assessing different size class of the same Diet analysis
fish species as each size class is assumed to play The diet was first determined by the frequency of
different ecological roles. This implies that food occurrence and percentage number method according
requirement at an early life stage of the same species to Hyslop (1980). Frequency of Occurrence accounts
can differ from the adult stage. Isumbisho et al., for the number of stomachs in which each prey item
(2004) stress the need to ascertain the feeding occurs and expressed as a percentage of the total
requirements of fish across its demographic in order number of stomachs examined
to provide a complete picture of resource use and
partitioning. The aims of this study were to elucidate Ji
seasonal and ontogenetic diet shifts and the feeding Frequency of Occurrence: Oi =
habits of H. vittatus in the Kalimbeza Channel on the
P
Where, Ji is number of fish containing prey i and P is
the number of fish with food in their stomach.

Feeding ecology of tiger fish, (Hydrocynus vittatus) in the Kalimbeza Channel
Number method was employed according to Zengeya (>176 mm). Length class selection was based on the
and Marshall, 2007. The number of individual prey per biology and life history of H. vittatus according to
stomach was enumerated and translated into a (Skelton, 2001). Data on prey items according to
percentage of the total number of prey items in the season and fish size were first checked for normality
specimen. Hence this was calculated as follows: and homogeneity of variances (Levenes test). To
improve on assumptions of normality and
homogeneity of variances, data were log10
Percent by number:
transformed. In case where transformation failed to
normalize the data, non-parametric Kruskal-Wallis
Where, Ni is the number of food category i analysis test was carried out to test the differences in
stomach contents between size classes while the
The food items were then combined into broader Mann-Whitney U-test was employed to test for
taxonomic categories for quantitative comparisons. differences in food items between wet and dry season.
The percentage of empty stomachs was determined. In A linear regression was employed to assess the
order to determine whether there was a change in relationship between the predator and prey length.
food composition among the different size groups, the Statistical analyses were performed using SPSS 24
fish were separated into three length class sizes ; small statistical software (Zar, 2010).
(<140 mm); medium (141-175 mm) and large
Figure 1. Map of the Zambezi Region (a), showing the Kalimbeza Channel on the Zambezi River (b), = gillnet
sites catchment areas. Generated using ArcGIS 9.3.

RESULTS was observed on day 16 denoted by the gape in the

graph. The number of empty stomachs observed
Diet composition of Hydrocynus vittatus between July and September were generally higher
A total of 498 specimens of Hydrocynus vittatus were compare to those observed between January and June
sampled during the survey. Out of the 498, stomachs (Figure 3).
dissected, 176 (35.3%) had food. Twenty-five different
prey items were identified and categorised into 18 The results shows that Synodontis spp (15.2%)
broad groups (Table 1). Of the fish prey items, 21.1% (17.8%) was the most important dietary component
could not be identified as they were in an advanced by percentage number and frequency of occurrence
stage of digestion. The major prey groups were; followed by Micralestes acutidens (10.3%) (12.1%),
Cichlids, Characids, Cyprinids, Mochokhids, Brycinus lateralis (7.8%) (9.2%) and Aquatic insects
Mormyrids, Micropanchax, Schilbids and Aquatic (21.1%) (7.5%) (Table 1). Other prey occurred less
Insects. Figure 2 shows the cumulative numbers of frequently, for instance, Petrocephalus catastoma,
unique prey species observed in the stomachs of H. Enteromius fasciolatus, Enteromius unitaeniatus,
vittatus over time. The results show a rapid detection Cyphomyrus cubangoensis, Petrocephalus catastoma,
of new prey species from day 1 up to day 28 and the Rhabdalestes maunensis and Schilbe intermedius
graph stabilizes between day 29 and 40. One or two contributed less than 5% to the total number of prey
new prey species were detected between day 41 and consumed (Table 1). Insects and fish were the two
42 and eventually the graph stabilizes for the next 48 major groups in the diet. Fish material were the most
sampling days and no new species were detected important food item and accounted for 80% of the
thereafter. Hence the sample size was considered total prey identified. Aquatic insects of the order
sufficient as no additional prey items could be Trichoptera and Odonata were the important groups
detected during the latter days of the sampling period of insect found in the stomachs of Hydrocynus vittatus.
(Mlanga, 2003) (Figure 2). No single stomach content
Table 1. Major prey categories and their importance in the diet of Hydrocynus vittatus in the Kalimbeza Channel
on the Zambezi River (February - December 2016). N = Total number while FO = Frequency of occurrence.
Prey items N %N FO %FO
Synodontis spp. 31 15.2 31 17.8
Micropanchax johnstoni 2 1.0 2 1.1
Enteromius lateralis 16 7.8 16 9.2
Insects 43 21.1 13 7.5
Enteromius fasciolatus 2 1.0 2 1.1
Enteromius poechii 7 3.4 7 4.0
Enteromius radiatus 6 2.9 6 3.4
Enteromius unitaeniatus 2 1.0 2 1.1
Cichlids 7 3.4 7 4.0
Marcusenius altisambesi 11 5.4 11 6.3
Cyphomyrus cubangoensis 1 0.5 1 0.6
Micralestes acutidens 21 10.3 21 12.1
Pharyngochromis acuticeps 4 2.0 4 2.3
Pollimyrus castelnaui 1 0.5 1 0.6
Petrocephalus catastoma 5 2.5 5 2.9
Rhabdalestes maunensis 1 0.5 1 0.6
Schilbe intermedius 1 0.5 1 0.6
Unidentified fish 43 21.1 43 24.7
Total 204 100 174 100

Figure 2. Cumulative number of unique prey items observed in the stomach of Hydrocynus vittatus in the
Kalimbeza Channel on the Zambezi River (February - December 2016).
Figure 3. Incidence of empty stomachs in 498 specimens of Hydrocynus vittatus in the Kalimbeza Channel on the
Zambezi River (February - December 2016).
Variation in relation to size class 2). Medium sized H. vittatus (141-175mm) had a total
Table 2 shows the variation in the food items of 14 different prey items in the stomachs. This group
according to the different size classes of H. vittatus. preyed heavily on Synodontis spp (24.7%), M.
The length of specimens sampled ranged from 58mm acutidens (17.3%) and Aquatic insects (15.8%). Specie
to 603 mm. The body weight also varied from 6.6 g to s such as E. fasciolatus, Enteromius radiatus, Enteromiu
2910 g. Out of the 176 H. vittatus specimens with s unitaeniatus and Pollimyrus castelnaui were observed
observable prey items in their stomachs, 69 samples in small quantities (Table 2). Large adults size class
were observed within the small size class, 59 in the (>176) showed a diet in which Synodontis spp (26.1%),
medium size and 44 in the large size class respectively. Brycinus lateralis (15.2%) and Micralestes acutidens
The small H. vittatus (less than 144 mm SL) size class (13.0%) dominated the diet composition. No
predominantly fed on Synodontis spp (14.1%), M. differences in diet were observed between small vs.
acutidens (9.9%) and Aquatic-insects (8.4%). medium size class, while significant differences were
Enteromius unitaeniatus, E. fasciolatus, Micropanchax observed between small vs. large and medium vs. large
johnstoni, Marcusenius altisa-mbesi, Pollimyrus size classes (p<0.05).
castelnauis were observed in small quantities (Table

Table 2. Diet composition (in percentage) of Hydrocynus vittatus in the Kalimbeza Channel from the Zambezi
River (February - December 2016) according to size class.
Prey items Small <140mm Medium 141- Large >176mm
(n=69) 175mm (n=59) (n=44)
Micropanchax johnstoni 1.4 2.5 0.0
Brycinus lateralis 5.6 9.9 15.2
Insects 8.4 15.8 4.4
Enteromius fasciolatus 1.4 2.5 2.2
Enteromius poechii 2.8 4.9 4.3
Enteromius radiatus 1.4 2.5 8.7
Enteromius unitaeniatus 1.4 2.5 2.2
Cichlids 4.2 7.4 4.3
Marcusenius altisambesi 1.4 2.5 2.2
Cyphomyrus cubangoensis 0.0 0.0 0.0
Micralestes acutidens 9.9 17.3 13.0
Pharyngochromis acuticeps 2.8 4.9 4.3
Pollimyrus castelnaui 1.4 2.5 0.0
Petrocephalus catastoma 2.8 4.9 2.2
Rhabdalestes maunensis 0.0 0.0 0.0
Schilbe intermedius 0.0 0.0 2.2
Synodontis spp. 14.1 24.7 26.1
Unidentified fish 40.8 71.7 8.7
Table 3. Percentage Frequency of occurrence in numbers of prey components of Hydrocynus vittatus in the
Kalimbeza Channel of the Zambezi River (February and December 2016) for the dry and wet seasons.
Species Wet Dry

FO %FO FO %FO
Rhabdalestes maunensis 0 0.0 1 1.1
Barbus fasciolatus 0 0.0 2 2.2
Marcusenius altisambesi 1 0.9 10 10.9
Micropanchax johnstoni 2 1.8 0 0.0
Brycinus lateralis 7 6.3 9 9.8
Cyphomyrus cubangoensis 1 0.9 0 0.0
Insects 11 9.8 32 34.8
Barbus poechii 4 3.6 3 3.3
Barbus radiatus 4 3.6 2 2.2
Pharyngochromis acuticeps 2 1.8 2 2.2
Pollimyrus castelnaui 0 0.0 1 1.1
Barbus unitaeniatus 2 1.8 0 0.0
Cichlids 2 1.8 5 5.4
Micralestes acutidens 17 15.2 4 4.3
Petrocephalus catastoma 0 0.0 5 5.4
Schilbe intermedius 0 0.0 1 1.1
Synodontis spp. 16 14.3 15 16.3
Unidentified fish 43 38.4 0 0.0
Total 112 100 92 100

Variation between seasons relationship. On average, H. vittatus consumed prey

The observed diet components of H. vittatus in the wet with a mean of 27.3% of its body length.
and dry seasons are presented in Table 3. Fish were
recorded in the stomachs more regularly among all
sizes in both seasons. A total of 112 prey items were DISCUSSION
recorded in the wet season while 92 were recorded in
the dry season (Table 3). During the wet season, H. Diet composition of Hydrocynus vittatus
vittatus consumed larger portions of Micralestes
acutidens (15.2%), Synodontis spp (14.3%), B. lateralis The stomach contents of many African inland water
(6.3%), and Aquatic insects (9.8%). The least fishes has been studied with a view to ascertaining
important prey items in the wet season were their dietary requirements in their natural habitats
Marcusenius altisambesi and Cyphomyrus cubangoensis. (Fagade and Olaniyan, 1973). Our results showed that
During the dry season, H. vittatus showed a shift to a Hydrocynus vittatus feeds extensively on non-cichlid
diet which was dominated by Aquatic insects (34.8%, fish species such as Synodontis spp. (17.8%),
mainly flying termites), Marcusenius altisambesi Micralestes acutidens (12.1%), Brycinus lateralis
(10.9%), Brycinus lateralis (9.8%) and Synodontis spp (9.2%) and Aquatic insects (7.5%). In a similar study,
(16.3%). The least important prey items were Mhlanga, (2003), reported that H. vittatus in Lake
Pollimyrus castelnaui and Schilbe intermedius. Kariba fed on Clupeidae (Limnothrissa miodon) (55%),
cichlids (20%), other fishes (including Clariidae
Predator Prey length relationship (catfish), Mochokidae (squeaker fish);
The prey length in relation to the predator length for (macroinvertebrates) (12%), and Alestiidae (Brycinus
H. vittatus is presented in Figure 4. Of the 126 spp.) (< 5%). Winemiller and Kelso-Winemiller,
Hydrocynus vittatus with measurable prey in their (1994) also showed that H. vittatus fed variously on
stomachs, the predator prey length relationship shows cichlids and Synodontis spp. (over 50%) in the Zambezi
that small H. vittatus ingested prey (fish) from small River floodplain suggesting that cichlids were the
size SL classes, whereas large H. vittatus ingested a major food source. Hence H. vittatus habitually feed on
wide range of prey sizes ranging from 50-170 mm SL diet components of animal origin (fin fish); the species
(Figure 4), and showed a positive predator prey is thus an exclusive piscivorous.
Figure 4. Predator prey length relationship for H. vittatus (Linear regression, P<0.05) at kalimbeza Channel,
Zambezi River.

The results of the stomach analyses affirm the Micralestes acutidens, Brycinus lateralis, Aquatic-
evidence of high incidence of empty stomachs as insects and cichlids; there are qualitative differences in
observed by Mhlanga, (2003), (Lake Kariba) and Dalu the items recorded and the dominant food items in the
et al., (2012) (Malilangwe reservoir). These findings diet of one particular size group match with those
are common in predatory fishes such as Tigerfish and which are of only minor importance in another size
the African pike. A high proportion of empty stomachs group. However, the major prey items which formed
in piscivorous fish can be explained by post-capture the main fish prey for H. vittatus includes Synodontis
digestion and change in daily temperature as spp. Micralestes acutidens and Brycinus lateralis which
postulated by He and Wurtsbaugh, 1993. The rate and are available in large quantities throughout the
change in temperature is directly proportional to the Zambezi River (Peel, 2012).
rate of digestion, with the emphasis that, the higher
the temperature, the higher the rate of digestion (He Seasonal shift in the diet of Hydrocynus vittatus
and Wurtsbaugh, 1993). Thus, fish which entangle gill Seasonally, all the diet components were encountered
nets at night would digest the food in their stomachs during wet and dry seasons throughout the study
faster under higher temperatures than cold gillnet period. However distinctive numerical variations in
nights (He and Wurtsbaugh, 1993). prey selectivity between the wet and dry seasons were
evident. A total of 112 prey items in wet season
relative to 92 prey items in dry season were recorded.
Evidence of ontogenetic diet shift in Hydrocynus
This imbalance may be explained by the abundance of
vittatus
prey items during the wet season and a decline in
There is evidence of ontogenetic diet shifts for the H.
prey items selection by H. vittatus in the dry season
vittatus where both small and medium size classes
(Winemiller and Winemiller, 1994). Similarly, the
feed on mixed proportions of both insects and fish
prominence of primary producers (e.g. algae, diatoms
relative to the large length class size which were
and zooplankton) during the wet season could be
typically piscivorous. These observations corroborated
linked to nutrient-rich white flood waters that
the report of Okon, (2002) on some aspects of the
stimulated greater primary and secondary
food and feeding habits of H.vittatus from a small
productivity (Winemiller and Winemiller, 1994). In
Malilangwe reservoir, Zimbabwe. The diets shift from
turn, primary producers such as macrophytes might
one particular food habits to another as found in this
have provided new habitats and served an alternative
study might also be explained by the period the diet
food source (Winemiller and Winemiller, 1994).
components are available in the resident water body
Variability in prey availability due to seasonal change
or the process of ontogenesis in organism (mouth size
has an indirect effect on the food composition of H.
gap) (Winemiller and Winemiller, 1994; Olojo et al.,
vittatus, implying that this species does not have a
2003). Keast, (1985a) postulated possible factors
strict food regime and this gives it a better chance of
which could account for producing size-related
survival. During the hot dry season characterised with
patterns of feeding among piscivorous fish. Firstly,
low water levels and dissolved oxygen, H. vittatus
small sized and juvenile fishes are inhibited by the
switched prey from Micralestes acutidens to Synodontis
small size of their mouth gape to exploit fairly small
spp (Winemiller and Winemiller, 1994). During this
prey items such as aquatic invertebrates and small
'crunch period', target preys become limited due to
prolific fish species such as barbs. Only after some
hostile shrinking water bodies (Winemiller and
time in growth will fish switch selecting small prey
Winemiller, 1994). During this period, the total
items to large prey items enhanced by an increase in
number of potential preys such as M. acutidens and B.
size of the mouth gape. Secondly, the observed
lateralis are reduced below sufficient levels to meet
switching from invertebrate to fish prey may be allied
the daily energy demand of the predator population
to changing prey availability at a particular point in
while hardy and tolerant species such as Synodontis
time as indicated by the seasonal prey availability
spp persist in bulk. These outcomes justifies that, the
Table 3. The occurrence of common prey categories in
observed seasonal differences in food composition of
all sizes also points to a likelihood of intraspecific
H. vittatus could be more derived from differences in
competition in H. vittatus. For instance, medium and
relative prey availability in the resident habitats rather
large H. vittatus in this study are likely to compete for
than prey selection by H. vittatus.
prey items such as Synodontis spp. Although there is
evidence of overlap among prey items such as

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RESEARCH ARTICLE
A Morphological Study of the Evolution of Eight Organisms

Through a Morphological Phylogeny
Sammer Marzouk

Received: 12.07.2017 Introduction: In this study, eight different shells from different species
Accepted: 15.08.2017 were examined by looking at the physical features of the shells. The
Published : 09.09.2017 purpose of the study was to identify evolutionary links between species
with shells. From these physical features, a morphological phylogeny was
Editor: compiled. And from the morphological phylogeny, it was shown that their
Dr. Arvind Chavhan the family of the shells was the first trait defined specific clades and that
each synapomorphic trait after this was a physical feature of the shells.
Sammer Marzouk (2017) A Results: From the experiment, the following conclusions were discovered:
morphological study of the evolution 1) That Scallops and Cockles are the most morphologic clay similar. 2) For
of eight Organisms through a
taxa A, trait III is an apomorphy because this traits is a single traits that
morphological phylogeny;
defines this taxa separately form the other taxa. 3) For taxa A, traits I is a
International J. of Life Sciences, 5 (3):
306-310. plesiomorphy because it is a trait that is ancestral to trait III which defines
taxa A.
Copyright: 2017| Author (s), This Conclusions: From the data, these conclusions could introduce a new topic,
is an open access article under the new ideas, and new information to existing researches. The influx of new
terms of the Creative Commons ideas would either become helpful, or build off the research of other
Attribution-Non-Commercial - No researchers. This would help the scientific community in increasing its
Derivs License, which permits use
knowledge of the evolutionary background of more species.
and distribution in any medium,
Keywords: Phylogenetics, Cladogram, Shells, Mollusk, Gastropod, Evolution
INTRODUCTION
On Earth, there may be more than two million species of organisms. These
organisms all share similar characteristics and some share common
ancestors. The group that was focused on in this study was organism's with
shells. These include the families of Mollusks and Gastropods. The purpose
of the study was to identify evolutionary links between species with shells
and shell groups that existed in the kingdom of life. The reason for
identifying evolutionary links between the evolutionary links between
these shelled organisms through phylogenies is that from this basic
phylogeny of only a few species, it can be expanded outward to hundreds of
other species. Eventually, an all encompassing phylogeny can be made with
all living organism in the animal kingdom

A morphological study of the evolution of eight Organisms through a morphological phylogeny
METHOD ancestor. In this paper, a mollusk is a group of animals

with soft body, no back bone and usually live in shells,
In this study, eight shells of different species origin and bivalves are shells that have two hinged parts, scallops
location were brought from the Great Lakes. These are a type of bivalve with a fan shaped shell with
shells were then sketched and analyzed. In the arches, cockles are a type of bivalve with turned end,
analyzation, the physical features of each of the cells conches are a type of mollusk with a spiral shell that
were noted and compiled. This became the basis for may bear long projections and have a flared lip,
the data matrix. In the data matrix, each specie with a gastropods is a shell that is univalve (has one hole),
specific trait was given a Y symbol, and each specie patellogastropoda is a group of organisms that live in
without a trait was given a N symbol. From the data cone shaped shells, length is when shell is put upright
matrix, the morphological phylogeny was compiled. In and measuring the right/left sides, body type 1 is a
the morphological phylogeny, traits in the data matrix spiral shell, body type 2 is a non-spiral shell, spiral
would be examined and species with specific traits spikes are pieces of shell on top that go around in a
would be in a clade, while species with other traits spiral pattern, flap is the piece of shell that connects
would not be placed in the clade. This would continue the shell to the outer covering, and incomplete/
until every species was in a separate taxon. After the complete refers to if the flap covers all of the opening,
construction of the morphological phylogeny, the taxa or just some of it (Fig. 2 ). The first shell given a Rough
and their placement would be analyzed for pattern and Scallop (Kurtz, 1860). It is white, black and gray and is
themes. 2.4 inches long (Fig. 3). The second shell given to us
was a trachycardium egmontiana or Florida Prickly
Cockle (Kurtz, 1860). It is 1.5 inches long and is a
RESULTS pearlescent color (Fig. 4). The third shell is a cone
shaped shell or a concolor polygramma (Kurtz, 1860).
The data used in the construction of this cladogram is It is white, black and gray and is 1 inch in length (Fig. 8
based on observation, and drawings of physical ). The fourth shell is the moonsnail shell (Kurtz, 1860).
features of the shells. These physical features must not This shell is white, is made up of layers and in 1 inch in
be able to be changed by outside forces, like sand and size (Fig. 5). The fifth shell is the Ribbed Cantharus
water and can be genetically traced back to a common Tranquebarica (Kurtz, 1860).
Table 1 Data Matrix

A B C D E F G H
Hinged Shell: Y Y N N N N N N
Non-hinged shell: N N Y Y Y Y Y Y
Free Swimming: Y Y N N N N N N
Bilaterally Symmetrical: Y N Y Y N N N N
Dark Colors: Y N Y N N N N N
Light Colors N Y N Y Y Y Y Y
Length Y Y N Y Y Y Y Y
Above 1 inch:
Shell is layered: N N N Y Y N Y Y
Smooth surface: N N N Y N Y Y Y
Spiral Spikes: N N N N N Y Y Y
Extended Piece of shell: N N N N N Y Y Y
Broken Piece at bottom: N N N N N Y Y Y
Symmetrical: Y N Y Y N N N N
(Line through top to bottom)
Arches: Y N Y N N N N N
True Coelom: Y Y N N N N N N
Hole on side: N N N N N N N N
Any Cracks: N N N N Y N N N
Symmetrical: Y N Y Y N N N N
(Line through top to bottom)

Fig. 1 :
Legend:
I = Animals with soft body, no back bone and usually live in shells
II = Shells that have two hinged parts
III = Type of bivalve with a fan shaped shell with arches
IV = Type of bivalve with turned end
V = Shell that is univalve (has one hole)
VI = Organisms that live in cone shaped shells
VII = Mollusk with a spiral shell that may bear long projections and have a flared lip.
VIII = Body Type 1
IX = Body Type 2
X = Bumpy Surface1
XI = Smooth Surface
XII = Spiral Spikes
XIII = No Swirl Flaps
XIV = Incomplete Flap
XV = Complete Flap
Numbers of Top:
C1 = Clade 1
C2 = Clade 2
Shell Pictures:
1Note: All of these physical characteristics are through genetics and are not caused by erosion of the shell from
natural forces like water and sand.

A morphological study of the evolution of eight Organisms through a morphological phylogeny
A= B=
Fig. 2 Fig. 3
C= D=
Fig. 4 Fig. 5
E= F=
Fig. 6 Fig. 7

G= H=
Fig. 8 Fig. 9
This shell is white and brown and has a size of 1.5 became a darker color. And example of this would by
inches and is layered with a bumpy surface (Fig. 6). the Spiky Florida Cocke or the Spiked Catharus (Table
The sixth shell is a Florida fighting conch shell (Kurtz, 1). We can also that the main difference between the
1860). It is a yellow and white color, with spiral spikes shells is if they are a bivalve of a gastropod (Fig. 1).
and is 2.2 inches long (Fig. 7 Shell F). The seventh shell The placing of the column was the first difference to
is a conidae, specifically a Malagasyconus lozet (Kurtz, divide up the Mollusc family (Fig. 1).
1860). This shell is a white, brown color and is 2
inches in length. The inside is a pink skin color (Fig. 8).
The eighth shell is a Lettered Olive (Kurtz, 1860). It is CONCLUSION
2 inches long, and is brown and white in appearance
(Table 1). The inside of the shell is white. In summary, the present data suggests that as time
progressed, shells started to gain more tetrapod-like
synapomorphies and became larger in size. Also, there
DISCUSSION was a divergence of becoming symmetrical and
radially symmetrical when the Gastropods and the
For clade C1, trait II is a synapomorphy of that clade Mollusks became two separate clades. Additional
(Fig. 1). The reason is because this clade made up of studies concerning the morphological and genetic
taxa A and taxa B is defined by the possession on this similarity between fish and shelled organisms might
trait II (Fig. 1). For taxa A and taxa B, trait I is a help better understand the link between gastropods,
symplesiomorphy because it is a common ancestral mollusks and fish.
trait to the synapomorphy of trait II that defines both
of these taxa. (Fig. 1) For taxa A, trait III is an Conflicts of interest: The authors stated that no conflicts of
apomorphy because this trait is a single trait that interest.
defines this taxon separately from the other taxa (Fig.
1). For taxa A, trait I is a plesiomorphy because it is a
trait that is ancestral to trait III which defines taxa A REFERENCES
(Fig. 1). From this set of data, we can that these
specific shells started out as a general family of Kurtz JD (1860) Catalogue of Recent marine shells,
Molluscs and then specialized due to the needed traits found on the coasts of North and South Carolina.
to survive in specific locations (Fig. 1). Those that had International Code of Zoological Nomenclature. 15,
natural predators that ate them would become spiky 4-9.
and ridged, and those who were living in darker areas 2017| Published by IJLSCI

RESEARCH ARTICLE
Effect of integrated use of lime, blended fertilizer and compost on

productivity, nutrient removal and economics of barley (Hordeum
vulgare L.) on acid soils of high lands in West Showa Zone of Ethiopia
Woubshet Demissie1, Selamyihun Kidanu2 and Cherukuri V Raghavaiah3*
1Soilfertility expert, Ministry of Agriculture, Addis Ababa, Ethiopia.

3Professor of Agronomy, Department of Plant Science, College of Agriculture and Veterinary Sciences, Ambo
University, Ambo, Ethiopia, East Africa.
*Corresponding author: Cherukuri V Raghavaiah, Email: cheruraghav@yahoo.in

Received: 28.02.2017 A field experiment was conducted during 2015 rainy season to investigate
Accepted: 04.05.2017 the effect of integrated nutrient and lime application on soil fertility and
Published : 09.09.2017 productivity of barley at Telecho, Wolmera district in the highlands of
Western Ethiopia. The experiment comprised nine treatments namely
Editor:
(t1):control, (t2):compost at 5t ha-1, (t3):lime at 611kg ha-1 , (t4) :compost
Dr. Arvind Chavhan
at 5t ha-1 + lime at 611kg ha-1, (t5): lime at 611kg ha-1 + DAP 150 kg ha-1
Cite this article as: and urea 72 kg ha-1research recommendation (60 kg n ha -1and 69 kg p2o5
Woubshet Demissie, Selamyihun ha-1), (t6):site specific fertilizer recommendation (npsb at 150 kg ha -1, kcl at
Kidanu and Cherukuri V Raghavaiah 100 kg ha-1, n at 33 kg ha-1), (t7): lime at 611kg ha-1 + npsb at 150 kg ha-1,
(2017) Effect of integrated use of kcl at 100 kg ha-1, n at 33 kg ha-1, (t8): lime at 611kg ha-1 + compost at 5t
lime, blended fertilizer and compost ha-1 + npsb at 150 kg ha-1, kcl at 100 kg ha-1, n at 33 kg ha-1, (t9): lime at
on productivity, nutrient removal 611kg ha-1 + 2.5 t ha-1 compost with 50 % of site specific fertilizer
and economics of barley (Hordeum
recommendation (npsb at 75 kg ha-1, kcl at 50 kg ha-1, n at 16 kg ha-1 ) laid
vulgare L.) on acid soils of high lands
out in a randomized complete block design with 3 replicates. Lime
in West Showa Zone of Ethiopia;
International J. of Life Sciences, 5 (3): requirement was determined based on exchangeable acidity. Soil analysis
311-322. revealed that lime application raised soil pH from 3.80 to the pH level
ranging from 6.63 to 6.86. Yield and yield components of barley was
significantly affected by the treatments. The highest and significant crop
yield response was obtained with t8 (5385.6 kg ha -1). Likewise nutrient
Copyright: 2017| Author (s), This uptake (n, p, k, s) was significantly higher in plots treated with t8 compared
is an open access article under the to t1, t2, t3, t4, t6. However, in most cases the difference in yield and yield
attributes, nutrient uptake, and soil parameters due to t1, t2, t3,t4, t6, were
not discernible suggesting that all elements of soil health and fertility issues
and distribution in any medium, should be addressed together. The highest net return (30633 birr ha -1)
provided the original work is with MRR (666.81) was obtained with t9. Thus, integrated use of lime at
properly cited, the use is non- 611kg ha-1 + 2.5 t ha-1 compost together with 50 % of soil test based
commercial and no modifications or fertilizer recommendation (npsb at 75 kg ha-1, kcl at 50 kg ha-1, n at 16 kg
adaptations are made. ha-1) for barley is advocated in the study area and other locations with
similar agro-ecologies to maximize barley grain productivity ha-1 and higher
economic return.
Key words: Lime, compost, soil pH, blended fertilizer, soil test based
fertilizer recommendation, barley yield, nutrient uptake, economics.

Woubshet et al., 2017
INTRODUCTION major constraints for small farmers to get the best out
of the input. Moreover the effectiveness of soil fertility
Barley (Hordeum vulgare L.) is the fifth staple food and interventions in Ethiopia has historically been
economically important crop in Ethiopia next to teff constrained by the lack of an integrated and locally-
(Eragrostis teff), maize (Zea mays L.), sorghum tailored approach.
(Sorghum bicolor) and wheat (Triticum aestivum) (CSA,
2009). The main advantage of incorporating barley in Thus, the current study was designed to assess the
diets oflate is due to its potential health benefits such influence of integrated use of lime, compost and
as lowering of blood cholesterol, with b-glucans blended fertilizers on productivity of barley under
(Behall et al., 2004), and the glycemic index (Cavallero acidic soil condition of West Shewa zone of Ethiopia.
et al., 2002) by barley has been reported widely (Pins
and Kaur, 2006).
MATERIALS AND METHODS
Even though barley represents about 13% of the total
national cereal production with an area of 1.04 million Description of the study area
hectares with an annual production of 1.7 million tons The experiment was conducted on acidic soils of
in main season (CSA, 2011), the problems of soil Welmera District in West Shewa Zone, Oromia
acidity and diseases decrease productivity of barley National Regional State Telecho kebele Farmers
and national average yield to as low as 1 ton ha-1 under Training Center (FTC) which is located at about 17 km
farmers condition in Ethiopia (Berhane et al., 1996; North of the district during the main rainy season
CSA, 2009). Besides, major production of barley still (June-November) 2015. Geographically, the district is
largely depends on the traditional varieties and located at 90 02' N Latitude and 380 34' E Longitude with
farming practices, which is also assumed to be one of altitude range from 2060-3380 m above sea level.
the constraints accounting for its low yield. Dega (41%) and Wainadega (59%) are the two agro
climatic zones of the district. Rainfall is bimodal: short
Most of the Ethiopian highlands are characterized by rains, belg, (March to April) and long rains, meher,
heavy rainfall and prone to soil acidity due to removal (June to September) (Welmera Agricultural office,
of ample amount of exchangeable heavy cations 2014).
through leaching, crop mining and runoff as compared
with grazing and forest lands. Hence, soil acidity is The annual rainfall, mean maximum and mean
now becoming a serious challenge for crop production minimum temperatures recorded for the year 2015
in the highlands of Ethiopia (Mesfin, 2007). According were 483.5 mm, 22.38 0C and 3.610C, respectively.
to the same author, soil acidity problem in Ethiopia is Woldeyesus (2005) reported, 85% of the total annual
mainly related to some of the Alfisols, and most rainfall is received between the months of June and
Oxisols and Ultisols soil classes that occur in the west, September and the rest from the end of March to the
north-western, south-western and southern parts of mid of May as a bimodal pattern. The soil type of the
the country. Currently, it is estimated that about 40% experimental site is Nitisols (Getachew et al., 2000).
of arable lands of Ethiopia are affected by soil
acidity/Al3+ toxicity (Taye, 2007). Soil acidity is Treatments and experimental design
expanding both in scope and magnitude in Ethiopia, The experiment was laid out in a randomized complete
severely limiting crop production which extend from block design (RCBD). Treatments and treatment
southwestern to northwestern with east-west combinations including the control treatment were
distribution but are concentrated in the western part assigned randomly to the experimental plots within a
of the country (Mesfin, 2007). Hence, Soil fertility block. The plot size was 3 m x 3 m (9 m2) and the crop
maintenance is a major concern in Ethiopia. Although was planted in rows with three replications. The
there is a gradual increase in the total volume of experiment consisted of nine treatments namely
fertilizers used in the country, low and unbalanced (T1)control, (T2)compost at 5t ha-1,(T3):lime at 611kg
application rates per unit area mainly focusing on ha-1 , (T4): compost at 5t ha-1 plus lime at 611kg ha-
1,(T5): lime at 611kg ha -1 plus DAP 150 kg ha-1 and
Urea and DAP fertilizers with low use efficiency of the
fertilizers (Getachew et al., 2009) and limited use of urea 72 kg ha-1research recommendation (60 kg N ha-
1and 69 kg P2O5 ha-1), (T6):site specific
improved seeds (Dercon et al., 2009) have remained

Effect of integrated use of lime, blended fertilizer and compost on productivity, nutrient removal
recommendation (NPSB at 150 kg ha -1, KCl at 100 kg and threshed on December 7 and 8, 2015 to determine
ha-1, N at 33 kg ha-1), (T7): lime at 611kg ha-1 plus grain yields.
NPSB at 150 kg ha-1, KCl at 100 kg ha-1, N at 33 kg ha-1,
(T8): lime at 611kg ha-1 plus compost at 5t ha-1 plus Data Collection
NPSB at 150 kg ha-1, KCl at 100 kg ha-1, N at 33 kg ha-1, Soil sampling and analysis
(T9): lime at 611kg ha-1 plus 2.5 t ha-1 compost with Prior to conducting the experiment, three
50 % of soil test based fertilizer recommendation representative composite soil samples were collected
(NPSB at 75 kg ha-1, KCl at 50 kg ha-1, N at 16 kg ha-1 ) from experimental site at plow depth of 0-20 cm on
laid out in a randomized complete block design April 15, 2015 and analyzed using standard laboratory
(RCBD) with 3 replications. procedures. Well decomposed compost sample was
analyzed for pH, organic carbon (OC), total N, available
Application of treatments and field management P, C/N ratio and exchangeable cations (Ca, Mg , K, Na ).
The experimental field was prepared by using oxen The soil samples were air-dried and ground to pass
driven local plow (Maresha) in accordance with through a 2 mm sieve. Soil pH (pH-H2O) and pH KCl
conventional farming practices followed by the (pH-KCl) was determined (1:2.5 soil to solution ratio)
farming community in the area. The field was plowed using a glass electrode attached to a digital pH meter
three times before planting, between the end of March (Page, 1982), soil texture was determined by
and the first week of June 2015. Lastly, the field was hydrometer method (Day, 1965). Methods as
leveled and divided into blocks which were then described by Walkley and Black for organic carbon,
divided into plots. and wet oxidation Kjeldahl method was used to
determine Total N, as described by Jackson (1967).
The field lime requirement was estimated based on Exchangeable bases were extracted with 1 M NH4O Ac
exchangeable acidity as follows at pH 7. Exchangeable Ca and Mg were measured from
the extract with atomic absorption spectrophotometer,
Lime requirement = EA*bulk density*0.15*area kg ha-1 while exchangeable K was determined from the same
2 extract with flame photometer. Exchangeable acidity
Thus, treatments that received lime, 611 kg ha-1 calcitic (Al and H) was determined from a neutral 1 N KCl
lime (CaCO3) was evenly broadcast manually and extracted solution through titration with a standard
mixed thoroughly in upper soils at 15 cm depth (plow NaOH solution based on the procedure described by
depth) in the first week of June 2015 as per the McLean (1965). Available P was determined using the
treatments. Compost was applied in the first week of Olsen extraction method (Olsen and Dean, 1965). In
June 2015 to all plots as per the treatments one month order to investigate the effect of liming to a change in
before planting. pH and electrical conductivity, composite soil samples
were again collected from each experimental plot on
The seeds of barley variety HB 1307 were sown by August 15, 2015 and were analyzed at National Soil
hand at the rate of 100 kg ha -1 on July 13, 2015 just Testing Center (NTSC). The pH was measured using
after one month of lime and compost application. A the same method as above, while electrical
Blended fertilizer, NPSB ( having 18.1 N- 36.1 P2O5- 6.7 conductivity was measured (1:2.5 soil to water ratio)
S-0.71 B nutrient ratio) applied at a rate of 150 kg ha -1 using EC meter Van Reeuwijk, (2002) and buffered
along with 60 kg K2O and 33 kg N ha-1 represents with KCl.
balanced nutrition recommended for barley
production in the study area (EthioSIS, 2014). While Agronomic parameters
treatments which receive Di ammonium Phosphate Data on days to 50% crop emergence, days to 50%
(DAP) applied at a rate 150 kg ha -1 along with 33 kg N . flowering and days to 50% physiological maturity
DAP, blended fertilizer NPSB; K2O were applied as were recorded when 50% of barley stands reached the
basal at time of planting. Nitrogen in the form of urea respective phenological stages. Phenological data were
was applied at tillering stage of barley 35 days after collected from the net plot area 2 m x 2 m (4 m 2) of
planting. The experimental field was hand weeded randomly selected five plants. Measurements of yield
twice at 25 and 45 days after planting to control attributes were taken at physiological maturity of the
weeds. At physiological maturity barley crop was crop prior to harvest. Plant height, number of tillers
harvested on 22 November 2015, sun dried for a while (effective tillers and total tillers) per plant, spike
www.ijlsci.in Int. J. of Life Sciences, Vol. 5(3) September, 2017 313

length and number of kernels per spike was recorded lime and compost were taken as 20 ETB 100 kg-1. The
from each plot. Plant height was recorded from ground cost of application and transport for fertilizer was
level to the top of the spike excluding awns. taken to be 15 ETB 100 kg-1.
The average yield was adjusted downward by 10%
The crop was harvested from the net plot areas and was used to reflect the difference between the
manually using sickle at the ground level and dry mater experimental field and the expected yield from
yield of the above ground biomass was determined. farmers fields with farmers practices from the same
Grain moisture content was determined and grain yield treatments (Getachew and Rezene, 2006). Analysis of
was adjusted to 12.5 % moisture content. Straw yield marginal rate of return (MRR) was carried out for non-
was recorded after uniform air drying of harvest for 3 dominated treatments, and the MRRs were compared
weeks. Harvest index was calculated as the percentage to a minimum acceptable rate of return (MARR) of
ratio of grain yield to the total above ground biomass 100% in order to select the optimum treatment.
yield. Thousand seed weight was determined by using
seed counter and weighting 1000 seeds sample taken Statistical Analysis
from each plot. The data were subjected to statistical analysis.
Analysis of variance (ANOVA) was carried out using
Plant Tissue Sampling and Analysis SAS statistical package program version 9.1.3. (SAS,
Grain samples at physiological maturity were collected 2002). Tukeys Test (P< 0.05) was used to assess
from each plot for determination of nitrogen, differences among treatment means where significant
phosphorus, potassium and sulphur concentration. differences were obtained by the analysis of variance.
The measurement of N was carried out according to Correlation analyses were determined through simple
the Kjeldahl procedure by transforming organic N into correlation coefficient between yield and yield
ammonium N by digesting with H2SO4 and a catalyst components.
(Chapman,1965). Nitrates and nitrites are initially
bounded by salicylic acid as nitro compounds and then
reduced by sodium thiosulfate. After digestion, the RESULTS AND DISCUSSION
solution was made alkaline by the addition of NaOH,
which allows the formation of volatile NH3. The Soil physical and chemical properties
measurement of P concentration of grain was carried The particle size distribution of the soil was dominated
out through calcinations of both grain and straw by clay fraction (58.47%) with 25.42 % silt and
separately at 450 C. After calcination, wet destruction 16.11% sand (Table 1). Getachew et al. (2003) found
of plant substances with strong acids was carried out, that the soil texture of the experimental site in
and then P was measured using dry ashing , Black, C.A Wolmera wereda prior to conducting the experiment
(flame Photometer) as described by Chapman (1965). was clayey in texture. The soil pH and exchangeable
The measurement of K and S in grain was carried out acidity (EA) of the experimental site were 3.80 (pH-
through Wet ashing-turbidity. The grain H2O) and 0.82, respectively. In most cases soils with
concentrations of N, P, K and S were used to estimate pH values less than 5.5 are deficient in Ca and/or Mg
the N, P , K and S uptake which was calculated by and also P (Marschner, 1995; Getachew and Sommer,
multiplying grain yields on hectare basis with the 2000). The soil reaction of the experimental site can be
respective N, P , K and S percentage . considered as extremely acidic (Landon,1991) which
suggests the presence of substantial quantity of
Economic analysis exchangeable hydrogen and aluminum ions which is
The market cost of barley was 8.50 Ethiopian Birr associated with acidity.
(ETB) kg-1. Field prices for Blended fertilizer (NPSB),
KCl , N and P from DAP and urea were taken as 13.65 The organic carbon and total nitrogen percentage of
ETB kg-1 , 14.50 ETB kg-1,. 14.00 ETB kg-1 and 11.50 the experimental field was 1.26 and 0.14, respectively
ETB kg-1 of nutrient, respectively. The field price of and rated as low as per Sahlemedhin (1999) and Bruce
lime and compost were taken to be 1.35 ETB kg-1 and and Rayment, (1982). For soil to be productive, it
0.50 EB kg-1, respectively. The costs of harvesting and needs to have organic carbon content in the range of
bagging were taken at 22 ETB 100 kg-1 of grain 1.8-3.0% to achieve a good soil structural condition
harvest. The cost of spreading and transportation of and structural stability (Charman and Roper, 2007).

Table 1. Selected physico-chemical properties of surface soil collected before lime application
Soil properties Values
Clay (%) 58.47
Silt (%) 25.42
Sand (%) 16.11
Textural class Clay loam
pH(H2O) 3.8
Organic carbon (%) 1.26
Total N (%) 0.14
Exchangeable acidity (cmolc kg-1) 0.82
Available P (ppm) 3.85
Table 2. Chemical composition of compost used in the experiment.

PH OC TN C/N Avail. Basic cations ( ppm kg-1 ) CEC
(H2O) (%) (%) Ratio P(ppm) (cmol(+)kg-1)
Ca Mg K Na
6.8 8.47 0.78 10.86 51.32 7245.6 806.69 819.56 52.91 44.62
OM= Organic matter, TN = Total nitrogen, C/N= Carbon to nitrogen ratio, P= Available phosphorus and CEC= Cation exchange
capacity, ppm = parts per million.
Table 3. Effect of liming on soil pH and electrical conductivity after two months of lime incorporation
Treatments pH EC (ds/m)
T1 4.16 0.07
T2 5.54 0.11
T3 6.63 0.14
T4 6.78 0.14
T5 6.82 0.16
T6 4.68 0.1
T7 6.71 0.14
T8 6.74 0.13
T9 6.86 0.11
EC = Electrical conductivity
T1- control; T2 - compost (5 t ha-1) ;T3-lime (611 kg ha-1); T4 - lime (611 kg ha-1) + compost(5 t ha-1);T5 - DAP(150 kg ha-1)
+KCl (100 kg ha-1)+ Urea (72 kg ha-1); T6-NPSB (150 kg ha-1)+ KCl (100 kg ha-1)+ Urea (72 kg ha-1); T7 Lime (611 kg ha-1)+
NPSB(150 kg ha-1)+ KCl (100 kg ha-1) +Urea (72 kg ha-1);T8 Lime (611 kg ha-1)+ compost (5 t ha-1)+NPSB (150 kg ha-1)+ KCl
(100 kg ha-1) +Urea (72 kg ha-1); T9 Lime(611 kg ha-1)+ compost (2.5 t ha-1)+NPSB(75 kg ha-1)+ KCl (50 kg ha-1) +Urea (36 kg
ha-1);
The result is agreement with the findings of Zelleke et Chemical properties of compost
al., (2010) who noted that the organic matter The organic carbon and total nitrogen contents of the
depletion is a wide spread problem in Ethiopian soils. compost is 8.47 and 0.78 %, respectively with the
Soil available P was 3.85 ppm, considered as low resultant narrow C:N ratio of about 10.85, indicating
(Berhanu, 1980). Available P of Alfisols (Nitisols) is low that the compost applied to experimental field is well
(Mesfin, 1998). Also Marschner (1995) stated in most decomposed. Brady and Weil (2002) recommends a
cases, soils with pH values less than 5.5 are deficient in C:N ratio of below 20 to have expected impact from
Ca and/or Mg, and also P. Thus, the fertility status of application of compost. The concentration of available
the experimental soil is sub-optimal for the production phosphorus was 51.32 ppm. The average
of barley. concentrations of basic cations, viz . K, Ca, Mg and Na
were 819.56, 7245.6, 806.69, and 52.91 ppm,

respectively, while the CEC was 44.62 cmol (+) kg -1.of experimental soils when lime requirement is
compost (Table 2) .The average pH (1:2.5 H2O) was 6.8 estimated based on exchangeable acidity, and thus
where most essential plant nutrients are available for indirectly favor the creation of more suitable medium
the crop. Getachew et al., (2012) reported similar for nutrient uptake by plants (Coventry et al., 1987).
result.
The result is in agreement with Bohn (2001), who
Lime requirement and its effect on soil properties stated that for agricultural purposes, soils with pH
The lime requirement of the experimental soil is 611kg values within the range of 5.8 to 7.5 are apt to be more
ha-1 and its effect on soil pH and EC measured after trouble free than those with higher or lower pH values.
two months of incorporation is presented in Table 3. Brady and Weil; (2002) and Rowell, (1994) also stated
Application of lime effectively increased the soil pH that lime application to acidic soils is one of the
from extremely acidic to medium and neutral range solutions to address soil acidity problem. Liming acid
(pH 6.63 to 6.86). Crop response to lime is principally soils has been suggested as the best method to attain a
a response to pH and the related secondary benefits suitable pH for growth of a variety of crops (Slattery
(Haynes and Ludecke, 1981). The results clearly and Coventry ,1993).
showed that liming visibly raise the pH value of the
Table 4. Effect of Integrated use of lime, blended fertilizer and compost on plant height, effective and total tillers,
spike length, number of kernels, Thousand seed weight, harvest index, grain yield, straw yield, biological yield and
straw yield of barley.
GY SY
BY
Treatments PH ET TT SL NK TSW HI (kg ha- (kg ha-
(kg ha-1)
1) 1)
T1 44.66c 2.33e 3.00d 5.70d 33.00b 9.92c 38c 1317.8c 3433.3c 2115.6b
T2 66.66b 3.66de 3.66cd 6.40c 34.66b 36.89b 39c 1617.0c 4173.3c 2556.3b
T3 63.00 b 4.00 cd 4.00 cd 6.03d 35.66 b 36.52b 37 c 1682.7 c 4483.3 c 2800.6b
T4 70.66b 5.33bc 5.33bc 6.56bc 37.66b 36.83b 40bc 1744.8c 4266.7c 2521.8b
T5 90.33 a 6.00 b 6.00 b 6.97a 45.66 a 38.44b 43 abc 3810.7 b 8916.7 b 5106.0a
T6 68.00b 3.66de 4.00cd 6.76ab 36.33b 36.59b 42abc 1670.2c 3966.7c 2296.5b
T7 92.33 a 9.66 a 9.66 a 6.90ab 48.33 a 37.69b 45 ab 4414.2 ab 9820.0 ab 5405.8a
T8 95.33a 10.66a 10.66a 7.03a 50.66a 44.02a 47a 5385.6a 11500.0a 6114.4a
T9 93.33 a 11.00 a 11.00 a 6.83ab 45.66 a 42.52a 44 ab 4799.6 ab 10766.7 ab 5967.0a
LSD (5%) 11.41 1.65 1.74 0.35 6.92 2 0.05 1099.7 2467.9 1465.3
CV (%) 5.24 9.22 9.55 1.89 5.92 2.25 4.43 13.08 12.66 13.21
Grand mean 76.03 6.25 6.37 6.57 40.85 35.42 42 2938.06 6814.07 3876
Means within a column followed by the same letter are not significantly different at 5% probability level.
PH-Plant height, ET-Effective tillers, TT- Total tillers SL-Spike length, NK-Number of kernels, TSW-Thousand seed weight, HI-
Harvest index, GY-Grain yield, BY-Biological yield, SY-Straw yield
T1- control; T2 - compost (5 t ha-1) ;T3-lime (611 kg ha-1); T4 - lime (611 kg ha-1) + compost(5 t ha-1); T5 - DAP(150 kg ha-1)
+KCl (100 kg ha-1)+ Urea (72 kg ha-1); T6-NPSB (150 kg ha-1)+ KCl (100 kg ha-1)+ Urea (72 kg ha-1); T7 Lime (611 kg ha-1)+
NPSB(150 kg ha-1)+ KCl (100 kg ha-1) +Urea (72 kg ha-1);T8 Lime (611 kg ha-1)+ compost (5 t ha-1)+NPSB (150 kg ha-1)+ KCl
(100 kg ha-1) +Urea (72 kg ha-1); T9 Lime(611 kg ha-1)+ compost (2.5 t ha-1)+NPSB(75 kg ha-1)+ KCl (50 kg ha-1) +Urea (36 kg
ha-1)
Yield and yield components imposed treatments (Table 4). The highest mean
height of 95 cm was recorded in barley plots received
Barley growth parameters T8- Lime (611 kg ha-1)+ compost (5 t ha-1)+NPSB (150
In the present study, plant height, measured at kg ha-1)+ KCl (100 kg ha-1) +Urea (72 kg ha-1) . In
physiological maturity, was significantly affected by contrast, control plots exhibited significantly lower

plant height compared to all other treatments. The wheat by applying manure and mineral fertilizer in
synergy between lime application and plant nutrition combination as compared to inorganic fertilizer alone.
was very conspicuous on response to height
increments of barley. Addition of lime with balanced Barley grain productivity
fertilization increased plant height by 24 cm compared Integrated application of lime with half and full
to balanced fertilizer sole application (68 cm). Plant recommended rate of organic and inorganic fertilizer (
height which was 60 cm with sole application of lime NPSB, urea, KCl and compost) gave many fold
increased to 90 to 95 cm when integrated with enhancement in grain yield over control plots. Hence,
balanced fertilizer. The result agrees with Mitiku et al., the highest mean yield of 5385.6 kg ha-1 was obtained
(2014) who showed a significant effect of combined over the control (1317.8 kg ha-1 ) with T8- Lime (611
application of organic and inorganic fertilizers on plant kg ha-1)+ compost (5 t ha-1)+NPSB (150 kg ha-1)+ KCl
height where the tallest barley plants were obtained (100 kg ha-1) +Urea (72 kg ha-1). This indicates that the
with application of 5 t ha-1 farmyard manure +75% of synergistic effect of applied lime in ameliorating soil
recommended NP at Adiyo and from the application of acidity along with recommended rate of organic and
5 t ha-1 vermicompost and 75% of recommended NP at inorganic fertilizers which gave greater productivity.
Ghimbo. Getachew (2009) also reported that the use of Integrated application of organic and inorganic
organic manures in combination with mineral nutrient sources was not adequate enough to increase
fertilizers maximized the plant height than the grain yield of barley significantly without integration
application of inorganic fertilizers alone. of lime (Table 4). Thus, the synergistic effect of lime
and plant nutrition made a significant difference in
The highest number of effective tillers (11) and total terms of grain yield increment. Application of lime
tillers (11) was obtained with T9- Lime(611 kg ha-1)+ with balanced fertilization increased barley grain yield
compost (2.5 t ha-1)+NPSB(75 kg ha-1)+ KCl (50 kg ha- by 2744 kg ha-1 compared to sole application of
1) +Urea (36 kg ha-1) which was comparable with T7- balanced fertilizer (1670 kg ha-1). On the contrary,
Lime (611 kg ha-1)+ NPSB(150 kg ha-1)+ KCl (100 kg grain yield which was 1.6 t ha-1with sole application of
ha-1) +Urea (72 kg ha-1) and T8- Lime (611 kg ha-1)+ lime enhanced to 4.4 to 5.3 t ha-1 when integrated with
compost (5 t ha-1)+NPSB (150 kg ha-1)+ KCl (100 kg ha- balanced fertilization. The lower yield exhibited in T6-
1) +Urea (72 kg ha-1). The attainment of higher number NPSB (150 kg ha-1)+ KCl (100 kg ha-1)+ Urea (72 kg ha-
of effective tillers might be due to the synergetic effect 1) (1670.2 kg ha-1) in the absence of lime integration
of lime, organic and inorganic fertilizers combinations was mainly due to adverse effect of soil acidity on the
that contributed to increased availability of NPKS. This availability of plant nutrients (Merino et al., 2010) .
result is consistent with Mitiku et al. (2014) who
observed that application of 5 t ha -1 farmyard manure The synergistic effect of applied lime with organic and
along with 75% of recommended NP gave highest inorganic plant nutrient sources highly influenced
number of productive tillers m-2 and highest number thousand seed weight of barley (Table 4). The highest
of grains per spike at Adiyo and Ghimbo ,respectively. thousand seed weight was recorded with T8- Lime
Spike length (SL) and number of kernels per spike (611 kg ha-1)+ compost (5 t ha-1)+NPSB (150 kg ha-1)+
(NK) of barley responded significantly to integrated KCl (100 kg ha-1) +Urea (72 kg ha-1) (44 g) followed by
use of lime and nutrients. The highest mean SL (7) and T9- Lime(611 kg ha-1)+ compost (2.5 t ha-1)+NPSB(75
KS (51) were obtained with T8- Lime (611 kg ha-1)+ kg ha-1)+ KCl (50 kg ha-1) +Urea (36 kg ha-1) (43g)
compost (5 t ha-1)+NPSB (150 kg ha-1)+ KCl (100 kg ha- over control plots (10 g) which is attributed to better
1) +Urea (72 kg ha-1) (Table 8). This showed an availability of nutrients and grain filling. The lowest
advantage of 23% and 53% increase in SL and KS, thousand seed weight in control plots could be due to
respectively as compared to control plots (Table 4). shriveled seeds that have small size which contributed
The possible reason for increased number of kernels to the less grain weight. A combination of lime, organic
per spike could be that mineral fertilizer and and inorganic fertilizers, increased thousand seed
mineralization of organic nutrient sources availability weight from 10 g to 44 g. Mitiku et al. (2014) also
due to effect of liming didnt put the plants to nutrient reported that application of 5 t ha-1 FYM in
stress at any stage throughout the growing season. In combination with 75% recommended rate of inorganic
agreement with this result Arif et al ., (2006) reported NP , the highest thousand barley grain weight was
significant increase in number of grains per spike of obtained at Adiyo as compared to application of 100%

recommended rate of inorganic NP which scored Ethiopia could be benefited by increasing grain yield
lower thousand grain weight. Similarly, Saidu et al., and straws to alleviate food and animal feed security
(2012) obtained the highest 1000 wheat grain weight, problems (Woldeyesus et al., 2004).
from application of 5 t ha-1 FYM in combination with
50% inorganic NP, while the lowest 1000 grain weight Nutrient concentration and uptake in barley
was recorded from no fertilizer application.
Integrated application of lime with organic and
Harvest index was influenced significantly with inorganic fertilizers significantly increased the
integrated application of lime together with concentration of TN %, P %, K meq/100 g and S % in
recommended rate of organic and inorganic fertilizers barley grain (Table 5). The combined effect of applied
. The highest harvest index (47) was obtained with T8- lime with organic and inorganic plant nutrient sources
Lime (611 kg ha-1)+ compost (5 t ha-1)+NPSB (150 kg significantly increased the nutrient concentrations of
ha-1)+ KCl (100 kg ha-1) +Urea (72 kg ha-1) as TN %, P %, K meq /100 g and S % from 1.16, 0.18,
compared to plots that received applied lime and 0.19, 0.17 to 1.37, 0.36, 0.28, 0.27, respectively as
lower doses of fertilizers (Table 4). In this study, compared to the control plots (Table 5). The linear
harvest indices ranged from 37 to 47 that showed an increment in N, P, K and S concentration with
increasing trend with increased combination of increased combination of applied lime along with
applied lime, organic and inorganic plant nutrient organic and inorganic nutrients in grain of barley
sources. The possible reason could be that application could be due to the effect of lime in ameliorating Al
of lime with increased rate of compost and inorganic toxicity and promoting root growth for nutrient
fertilizers might have increased the efficiency of barley uptake coupled with nutrient availability from the
to partition the dry matter into the reproductive seed synergistic effect of both applied organic and inorganic
sinks. Mooleki et al., (2002) too reported that the nutrient sources.
increased rate of either FYM or inorganic NP has
increased the harvest index of rice. Likewise, a distinct variation in TN, P, K and S uptake
in grain was observed due to different integrated use
In line with this, Ivarson (1997) noted that effect of of lime with organic and inorganic fertilizers. The
liming attributed to a reduction of aluminum toxicity highest removal of N (74 kg ha-1), P (19.65 kg ha-1) , K
and an increase in soil pH that can both result in an (16.05 kg ha-1) and S (15 kg ha-1) in grain was
increase of microbial activity and release of labile obtained with T8- Lime (611 kg ha-1)+ compost (5 t ha-
organic matter (Anderson 1999). Furthermore, 1)+NPSB (150 kg ha-1)+ KCl (100 kg ha-1) +Urea (72 kg
Shiferaw and Anteneh, (2014) found that application ha-1) as compared with control . The nutrient uptake
of lime and all combinations of fertilizers, either alone increased through application of lime and compost
or combined, significantly increased barley yield over with blended macronutrients and micronutrients in
untreated control. appropriate form of fertilizer to nutrient deficient soil.
Hence, the complimentary effects of applied lime,
The highest total biomass yield (11500 kg ha-1) and organic and inorganic fertilizer (NPSB, urea and KCl
straw yield (6114 kg ha-1) were obtained with T8- and compost) ultimately lead to more nutrient uptake.
Lime (611 kg ha-1)+ compost (5 t ha-1)+NPSB (150 kg The N losses due to leaching or denitrification might
ha-1)+ KCl (100 kg ha-1) +Urea (72 kg ha-1) as have been reduced in soil by mixing N fertilizer with
compared to control (biomass3433 kg ha .and straw
-1 organic compost, resulting in better utilization of N by
yield 2115 kg ha-1). This is in agreement with plants. Composted organic materials enhanced
Agegnehu et al., (2014) who found that application of fertilizer use efficiency by releasing nutrients slowly
half the recommended NP rate and half recommended and thus reducing losses, particularly of N (Ramos and
rate of manure and compost as inorganic N Marttinez Casasnovas,2006). This may be attributed
equivalence resulted in wheat grain and total biomass to the improvement of soil physical properties by
yield advantages of about 129% and 194% compared organic nutrient sources in addition to contributing to
to the control. Therefore, this condition is quite nutrient availability (Laila and Ali, 2011). This result
important that increasing both parameters (straw and corroborates with the findings of Nasreen and Farid
total biomass yield) without affecting the grain yield (2003) who demonstrated that the application of
through lodging effect, farmers in the highlands of

Table 5. Effect of Integrated use of lime, blended fertilizer and compost on total nitrogen,
phosphorus, potassium and sulphur concentrations and uptake (Kg ha -1) in barley grain
Nutrient concentration Nutrient uptake(kg/ha)
Treatments
TN% P% K(meq/100g) S% TN P K S
T1 1.16f 0.18d 0.19c 0.17d 15.29b 2.46c 2.59b 2.25c
T2 1.21e 0.22c 0.21bc 0.18cd 19.67b 3.55c 3.52b 2.95c
T3 1.22e 0.22c 0.22bc 0.18cd 20.53b 3.69c 3.70b 3.01c
T4 1.25d 0.23c 0.24b 0.19cd 21.91b 4.04c 4.18b 3.36c
T5 1.28cd 0.32b 0.27a 0.20c 60.91a 15.34b 12.95a 9.82b
T6 1.22e 0.23c 0.23b 0.18cd 20.37b 3.83c 3.84b 3.12c
T7 1.32b 0.33b 0.28a 0.26ab 58.56a 14.56b 12.70a 11.47b
T8 1.37a 0.36a 0.28a 0.27a 74.15a 19.65a 16.05a 15.13a
T9 1.31bc 0.32b 0.27a 0.25b 63.19a 15.51b 13.27a 12.00ab
LSD (5%) 0.034 0.03 0.03 0.02 15.994 3.39 4.25 3.62
CV (%) 0.95 4.38 4.44 4.53 14.18 12.9 18.38 18.06
Grand mean 1.26 0.27 0.24 0.21 39.4 9.18 8.09 7.01
Means within a column followed by the same letter are not significantly different at 5% probability level.
TN-Total nitrogen, P- Phosphorus, K-Potassium ,S = Sulphur
T1- control; T2 - compost (5 t ha-1) ;T3-lime (611 kg ha-1); T4 - lime (611 kg ha-1) + compost(5 t ha-1); T5 -
DAP(150 kg ha-1) +KCl (100 kg ha-1)+ Urea (72 kg ha-1); T6-NPSB (150 kg ha-1)+ KCl (100 kg ha-1)+ Urea
(72 kg ha-1); T7 Lime (611 kg ha-1)+ NPSB(150 kg ha-1)+ KCl (100 kg ha-1) +Urea (72 kg ha-1);T8 Lime (611
kg ha-1)+ compost (5 t ha-1)+NPSB (150 kg ha-1)+ KCl (100 kg ha-1) +Urea (72 kg ha-1); T9 Lime(611 kg ha-
1)+ compost (2.5 t ha-1)+NPSB(75 kg ha-1)+ KCl (50 kg ha-1) +Urea (36 kg ha-1).
different nutrients caused significant increase in The highest net return of 30633.00 Ethiopian Birr with
nutrient uptake by pea. Uptake of N, P, K, S and Zn by highest MRR value of 666.81 was obtained from T9-
shoot and seed was highest with the treatment 30 kg Lime (611 kg ha-1)+ compost (2.5 t ha-1)+NPSB (75 kg
N, 50 kg P, 40 kg K and 20 kg S ha -1. Furthermore ha-1)+ KCl (50 kg ha-1) +Urea (36 kg ha-1) as compared
Sawer et al. ( 2009) stated that the nutritional status of to T5 - DAP(150 kg ha-1) +KCl (100 kg ha-1)+ Urea (72
plants was further strengthened when chemical kg ha-1) with net return of 24488.00 ETB and MRR
fertilizer was combined with compost. 381.00 (Table 6). This means for T9 on an average for
each l Birr ha-1 invested, the return was 1 Birr, plus
Economic feasibility analysis 6.00 birr ha-1 in net benefit which is economically
feasible as compared to T5 that showed 1 birr
Field price of barley, lime and compost were 8.50 birr recovery plus 3.81 birr ha-1 net benefits. This
kg -1, 1.35 birr kg-1 and 0.50 birr kg-1, respectively, recommendation is also supported by CIMMYT (1988)
while field price of DAP, Urea, NPSB and KCl were which stated that farmers should be willing to change
14.00 birr kg -1 11.50 birr kg -1 ,13.65 birr kg -1and from one treatment to another if the marginal rate of
14.50 birr kg -1 ,respectively. The cost of harvesting return of that change is greater than the minimum
and bagging were taken as 22 birr 100 kg -1. The cost acceptable rate of return. The current study suggests
of incorporation and transportation of lime taken as that it would be advisable for farmers in the study area
20 birr 100 kg-1.. The cost of incorporation and to apply integrated lime at 611kg ha-1 plus 2.5 t ha-1
transportation of compost taken as 10 birr 100 kg -1 compost with 50 % of soil test based fertilizer
(the cost of transport for compost was minimum since recommendation (NPSB at 75 kg ha-1, KCl at 50 kg ha-1,
it was purchased in the vicinity of farm), the cost of urea at 36 kg ha-1 ) for enhancing barley grain yield ha -
1 and accruing highest economic return.
application and transport for fertilizer was taken to be
15 birr 100 kg1

Table 6: Economic analysis of Integrated use of lime, organic and inorganic fertilizer in terms of partial budget
and marginal rate of return (MRR) for barley production
Trt . Grain Adjusted Gross return Total variable Net return MRR (%)
No yield(kg/ha) yield-10% (ETB ha-1) cost (ETB (ETB ha-1)
1 1317.77 1185.99 10080.9 260.92 9820.02
3 1682.69 1514.42 12872.6 1280.22 11592.4 173.92
2 1616.99 1455.29 12370 3320.16 9049.79D -
4 1744.82 1570.34 13347.9 4292.52 9055.36D -
5 3810.67 3429.61 29151.7 4662.86 24488.8 381.25
6 1670.2 1503.18 12777.1 4704.5 8072.55D -
9 4799.64 4319.67 36717.2 5584.28 30633.00 666.81
7 4414.24 3972.81 33768.9 6194.87 27574.00D -
8 5385.59 4847.03 41199.8 9387.2 31812.6 31.01
D: Stands for dominated treatment.
CONCLUSION test based fertilizer recommendation (75 kg ha -1NPSB,

50 kg ha-1KCl, 36 kg ha-1 urea) accrued the highest net
From the foregoing account it could be concluded that return of 30633.00 ETB ha-1with MRR of 667% which
application of lime (611 kg ha-1) increased the soil pH is advisable for farmers to maximize barley grain yield
of extremely acidic soil (pH 3.8 ) to medium and ha-1 and economic return on acid soils of central high
neutral range (pH 6.63 to 6.86) through its lands of Ethiopia where soil acidity is a major
ameliorating effect and thus indirectly favouring the production constraint.
creation of more suitable medium for nutrient uptake
by the crop. This condition creates a favorable soil Conflicts of interest: The authors stated that no conflicts of
environment that enables efficient use of both organic interest.
and inorganic nutrients which ultimately resulted in
better performance in terms of yield and yield
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RESEARCH ARTICLE
Seasonal distribution and abundance of Thrips (Thysanoptera:

Thripidae) on onion producion in central zone of Tigray, Ethiopia
Gebretsadkan Zereabruk
Tigray Agricultural Research Institute, Axum Agricultural Research center, P. O. Box: 230, Axum, Ethiopia; Email:
gebretsadkanz2007@gmail.com,
Received: 27.04.2017 Knowledge of insect pest population dynamics and abundance can help to
Accepted: 05.06.2017 predict when and where the highest infestations will occur. This
Published : 09.09.2017 information is very important to apply the right pest control measures at
the right time and place. Therefore, a field survey was conducted in four
onion producing districts namely, Ahferom, Mereb-leke, Adwa and Laelay-
Editor: maychew from 2015-2016 to assess the distribution, abundance and
Dr. Arvind Chavhan significance of thrips on onion production. Survey results indicated that
only onion thrips (Thrips tabaci) was found in all study areas. Thrips were
found to be widely distributed on onion crops in all of the areas surveyed
Gebretsadkan Zereabruk (2017) and were important insect pests of the crop. Onion thrips abundance varied
Seasonal distribution and abundance according to growth stage of the plant, months, locations (altitudes) and
of Thrips (Thysanoptera: Thripidae) weather factors mainly (temperature and rain fall). The number of onion
on onion producion in central zone of thrips per plant was low in the early December however; it was
Tigray, Ethiopia; International J. of progressively increased to the peak population in March. The highest pest
Life Sciences, 5 (3): 323-331. load of thrips population on onion was observed during March at Mereb-
leke, Ahferom, Adwa and Laely-maychew with 340, 270, 266 and 227 thrips
per plant, respectively. The highest number of onion thrips was recorded at
Copyright: 2017 | Author (s), This lowland areas (1386 m.a.s.l.) at Rama, which were characterized by sandy
is an open access article under the to loam soil type and hot weather conditions. Onion thrips population was
terms of the Creative Commons positively correlated (r = + 0.94) with increasing temperature and
Attribution-Non-Commercial - No negatively correlated (r = - 0.93) with relative humidity. At higher
Derivs License, which permits use infestation, thrips caused up to 50-75% yield loss and others observed 30-
and distribution in any medium, 50% yield reduction when control measures are not taken 27.5 and 50% of
provided the original work is the respondents, respectively. These results are important for the
management of onion thrips by growers.
adaptations are made. Key word: Onion thrips, onion, abundance and farmers
INTRODUCTION
Onion thrips is originated in the Mediterranean region. It was first found in

Hawaii in 1915, currently it is found all over the world where onion is
grown (Brian, 2006). They are poor fliers (MAFF, 1997); however their
feathery wings allow them to be readily carried by air currents. In the
tropics, thrips population on annual crops are possibly affected more by

Gebretsadkan Zereabruk, 2017
immigration than in north temperate regions, due to results in leaf tissue silvering and photosynthesis
the north-south movement of inter tropical reduction, leading to bulb size reduction and yield loss
convergence zone. Thrips can colonize crops from sea (Carl, 1997). Damaged leaves may become papery and
level up to 2000 m above sea level (Mishra et al., distorted. Infested terminals lose their color, roll, and
2014). In Ethiopia, the particular insect is also tiny black tar spots of thrips excrement are evident
distributed throughout the country, particularly in on leaves with heavy feeding injury (Diane, 2008).
onion growing areas (Tadele and Amin, 2014).
Feeding on leaves also can create an entry point for
The rate of development of Thrips tabaci is positively plant pathogens (Orloff et al., 2008). In addition to the
affected by increased temperature and decreased by direct damage, thrips cause viral and fungal diseases
increased relative humidity (Hamdy et al., 1994). In on onions under field and storage conditions. It is an
addition to their effect on thrips activity, temperature important vector that is an organism that transmits
and relative humidity further influence the intrinsic infection from one host to another. Both adults and the
rate of natural increase of the thrips (Murai, 2000). wingless larvae are attracted to white, yellow and
other light colored blossoms and are responsible for
Onion thrips damage to young onions is more spreading tomato spotted wilt virus (TSWV) and
devastating than on larger plants late in the growing impatiens necrotic spot virus. Plant pathogens
season (Zsofia and Adam, 2012). Thrips prefer to feed penetrate the injured plant easily. Alternria porri Ellis
on the newly emerged leaves in the center of onion penetrate the surface of plant pierced by thrips and
necks. Under crowded conditions, they will move cause necrotic spot on the leaves and large population
toward leaf tips to feed. Both adult and larval thrips of thrips increase the incidence of fungal pathogen in
feed within the mesophyll layer using a punch-and the field (Rueda and Shelton, 2003). Thrips tabaci
suck motion. Older leaves that have been folded over transmits purple blotch on onion (Mckenzie et al.,
may also be a preferred feeding site for the pest 1993; Carl, 1997). Similarly, the infected bulbs in the
(Paibomesai et al., 2013). Onion thrips consume field cause Botrytis allis (Hann) disease in stored
mesophyll cells, which eventually results in a loss of onions (Mayer et al., 1987). Thrips are the only vectors
chlorophyll and reduced photosynthetic efficiency of tospo viruses and thrips tabaci, in particular, is the
(Boateng et al., 2014). The mouthparts are beak- vector of Iris yellow spot virus (IYSV), genus Tospo
shaped with one enlarged mandible (tooth). The beak virus (Bunyaviridae), a severe and widespread disease
and mandible is thrust forward to puncture the leaf infecting onion, leek, iris and wild Allium species.
epidermis and sap released from injured plant cells is Symptoms of iris yellow spot include dry, elongated,
sucked up. Removal of chlorophyll causes the feeding straw-colored to tan or white lesions on onion leaves
area to appear white to silvery in color. Areas of leaf and scapes (the flowering stems of bulb plants). Small
injury can occur as patches and streaks (Diane, 2008). lesions may resemble thrips-feeding injury (Tilm and
When damage is severe, these small patches can Carrie, 2014). Recently, onion thrips was shown to
occupy most of the surface of the leaf and the plant transmit a bacterial pathogen Pantoea ananatis (center
cannot adequately photosynthesize and nutrient rot) to onion (Dutta et al., 2014). P. ananatis causes
transportation to the bulb is reduced. Yield reductions center rot in onion and has caused substantial
in the form of reduced bulb size is the primary effect of economic losses in the United States.
onion thrips (Rueda and Shelton, 2003; Cranshaw,
2004). In Ethiopia, it is an important insect pest that affect
onion yield by direct feeding as well as reducing the
Onions are most sensitive to thrips injury during the quality and quantity by rasping the leaves and other
rapid bulb enlargement phase that occurs in dry tissues of onion crops to release the nutrients
season, particularly during irrigation season in (Tsedeke, 1985). Onion fields can be destroyed by
Ethiopia (Tsedeke, 1985). Accelerated plant maturity onion thrips, especially in dry seasons (Tadele et al.,
and senescence due to thrips injury may truncate the 2013). Tsedeke (1985) and Yeshitila (2005) reported
bulb growth period. Following harvest and during that onion bulb yield losses of 33 and 26-57%,
storage, thrips may continue to feed on onion bulbs, respectively, due to onion thrips in Ethiopia. Similar
causing scars that reduce quality and aesthetic studies at Upper Awash Agro Industry Enterprises
appearance of bulbs. Thrips tabaci feeding damage revealed yield losses of 10 to 85% due to onion thrips

Seasonal distribution and abundance of Thrips (Thysanoptera: Thripidae) on onion producion
in Ethiopia (Bezawork, 2006). In the study area even select the farmers for the interview based on their
though the extent of damage and their abundance on area coverage of onion production i.e. more number of
onion has not been studied earlier, it is the most farmers was interviewed in the highest production
important pest of onion in the region. Knowledge of areas and less number of farmers in low production
insect population dynamics and abundance can help to areas. Questionnaires focused on constraints
predict when and where the highest infestations will (concerning insect pests and diseases), cropping
occur, how much the pest load will become and for season, history of thrips in the study area, season of
how long they will stay. This information is very the year when thrips observed in onions, local name of
important to apply the right pest control measures at thrips, thrips intensity on onions, other crops attacked
the right time. Therefore, the objective of the study by thrips in the area, changes occurred since known,
was to assess the distribution, abundance and cropping months at which thrips are serious,
significance of thrips on onion production across management practices, type and frequency of chemical
duration and locations. application.
MATERIALS AND METHODS RESULTS AND DISCUSSION
Distribution and abundance of onion thrips Abundance and Distribution of thrips on onion
The distribution of thrips was assessed on irrigated Onion crops are attacked by various thrips species. In
onion fields of four onion producing districts in Ethiopia the common species of thrips involved in
Central Zone of Tigray Region namely Ahferom, Adwa, onion infestation were Thrips tabaci Lind and
Mereb-leke and Laelay-maychew. These areas vary Frankliniella occidentalis (Pergand) However, Thrips
from each other, in elevation (Table 3), temperature, tabaci (Thysanoptera: Thripidae) was the only species
soil type and crop cultivation practices. A field survey found in all the surveyed areas. Results indicated that
was conducted monthly in the dry season on irrigated onion thrips abundance varied according to growth
onion fields from December 2015 to April 2016. stage of the plant, months, locations (altitudes) and
Depending on availability of onion fields, fields along weather factors mainly (temperature and rain fall)
the main road were considered and visited for (Figure 1). During the sampling period low number of
sampling. Up to ten fields were sampled per wereda by thrips were observed during early seedling stage and
sampling fields at 5-10 km interval along the main increased gradually through time with growth and
road. Ten plants per field were sampled randomly and development of the plant. In most of the surveyed
inspected for thrips. Samples were taken in a zigzag areas, less number of thrips per plant was observed
fashion. Thrips on each of the ten plants were counted during early December up to January. Most of
using 10x magnifying hand lens. The mean number of producers transplanted their onion seedlings in these
thrips was calculated per plant per month in each months. Similarly, at seedling stage (2-3 leaf stage) in
location. nursery sites a few numbers of thrips (1-2 thrips per
plant) were recorded. As far as the locations are
The mean number of thrips from December to April concerned, the highest (61) and the lowest (37)
was used to determine the abundance of thrips on numbers of thrips per plant were recorded at Mereb-
irrigated onion fields. Up to twenty thrips per field on leke and Laely-mychew, respectively in December. The
each sampling occasion was collected to determine the highest pest load of thrips population on onion was
species of thrips. Adult thrips were identified using a observed during March at Mereb-leke, Ahferom, Adwa
stereo microscope in laboratory at Axum University. and Laely-maychew with 340, 270, 266 and 227 thrips
Sparks et al. (1914) key was used for identification. per plant, respectively (Figure 1). This could be due to
the dry weather condition of that period, which helped
Farmers perception the buildup of thrips population. Thrips were found to
The study was conducted in four weredas namely be widely distributed on onion crops in all of the areas
Ahferom, Adwa, Mereb-leke and Laelay-maychew by surveyed and were important insect pests of the crop.
interviewing forty farmers from each using a
structured questionnaire. Reports of Agriculture and The highest 340 onion thrips per plant was recorded
Rural Development office of the weredas were used to at Mereb-leke lowland areas at Rama, which were

characterized by sandy to loam soil type with 1386 onion crop was very low during seedling stage and it
m.a.s.l. As compared to Mereb-leke, a lower number of was gradually increased during vegetative stage of the
thrips per plant (227) was recorded at Laelay- crop and reached a peak occurrence during
maychew with clay soil type with 2131 m.a.s.l. At physiological maturity stage. Tsedeke (1986) indicated
Adwa and Ahferom thrips distribution was similar. At that, thrips population remained low from June
the end of March and early April there was unusual through August and started rising in September and
rainfall in all surveyed areas. As a result the thrips reached its peak in April. Thrips population change
densities per plant decreased to low numbers per mainly brought by the environmental factors such as
plant. Farmers in the survey areas had different temperature and rainfall. Similarly, from dry and rainy
management practices; some of them used good period observation in Indonesia, Vos et al. (1991)
agronomic practices and others were poor in their reported that thrips population was severe in dry and
management. In onion fields with poor managements, hot periods than the rainy season. This was due to
such as weedy fields, less irrigation water and rarely wash off thrips by rainfall and the favorable humid
on saline soils, higher thrips population per plant was conditions created for entomo-pathogenic fungi that
observed. On weedy onion fields with higher thrips feed on thrips. Different people reported the impacts
infestation, symptoms of Iris yellow spot virus (IYSV) of climatic factors on thrips. Krantz et al. (1978)
were observed on onion leaves at Ahferom and Mereb- indicated that, the number of thrips on onion crop
leke. Similarly, at Laelay-maychew and Ahferom there increased rapidly during dry weather and decreased
was a problem of purple blotch and white rot diseases. rapidly after rain. Heavy rainfall greatly reduces the
When the plant reaches at good vegetative stage (7-10 population of thrips by both washing from the plant
leaves) there was a symptom of leaf blight (suddenly and killing those which are in the soil (Cranshaw,
the leaves become brown and died) at Mereb-leke and 1994). Similarly, Workman and Martin (2002)
Laely-maychew. reported that damage and density of thrips on crops
were heavier in dry and hot seasons due to their high
This study was in agreement with the results reported reproduction rate and short developmental periods at
by different researchers. Mallinath et al. (2014) such weather conditions and heavy rainfall could
reported that the population of thrips occurrence on cause up to 70% thrips mortality.
Figure 1. Onion thrips abundance across locations and months of growing season

Table 1. Farmers responses to questionnaires during survey about thrips in onion production
No. Questioners Variables Respondents (%)
1 Onion production season 1. Irrigation 100
2. Rainfed -
2 Onion production constraints regarding pests 1. Onion thrips 80
2. Diseases 20
3 Observation of thrips on other crops 1. Yes 22.5
2. No 77.5
4 Other vegetables infested by thrips 1. Tomato -
2. Garlic 22.5
3. Cabbage -
5 Duration of thrips known 1. Five years 27.5
2. Eight years 40
3. Ten years 32.5
6 Changes observed on thrips intensity over years 1. Increase 100
2. Decrease -
3. No change -
7 Months with the highest thrips population 1. March-April 47.5
2. Jan-Feb. 32.5
3. Oct-Dec 20
8 Onion yield loss due to thrips (%) 1. 20-30 22.5
2. 30-50 50
3. Above 50 27.5
9 Onion thrips management practices 1. Chemical 80
2. Cultural -
3. No control 20
10 Type of insecticide used in thrips management 1. Karate 5%Ec. 35
2.Malathion 50% 30
3.Any insecticide 15
11 Frequency of spray interval 1. once in a week 20
2. 3times a month 35
3. If necessary 25
12 Efficacy of insecticides on thrips management 1. Very effective -
2. Moderately 3.1
3. Very low 56.2
4. Not effective 40.6
13 Price of insecticides concerning producers 1. Affordable -
2. Not affordable 80
Farmers perception about onion thrips not taken 27.5 and 50% of the respondents,
Farmers responses to the questionnaires are shown in respectively. The majority of the farmers (80% of
(Table 1). Farmers recognized that onion production is them) indicated that they depend on insecticides to
affected by insect and disease problems 80 and 20% of manage onion thrips. The frequency of insecticide
the respondents, respectively. Farmers locally name application was reported to vary from once per week
onion thrips as kunchi shigurti or hafo shigurti in to three times per month. However, some of the
Tigrigna. According to farmers responses the highest respondents (25%) responded that they apply when
thrips infestation was observed from March to April they observed higher infestations or if it deemed
(47.5% of respondents). At higher infestation, thrips necessary. The commonly used chemical insecticides
caused up to 50-75% yield loss and others observed were karate 5% EC (lambda-cyhalothrin) and
30-50% yield reduction when control measures are Malathion 50% EC. The effectiveness of the chemicals

was very low to non effective according to 56.2 and years. It may mean that the pest started causing heavy
40.6% of the respondents, respectively. The prices of damage to the crop in the last few years. The reason
the chemicals were also variable and not affordable by for the increase of thrips problem on onions could be
the onion producers. many, but production system changes from rainfed to
irrigation, climate change effects, use of high yielding
Farmers were asked whether they observed thrips on improved varieties that lack the resistance genes to
other crops in their fields and the majority (80%) of thrips, and regular unwise use of insecticides could be
them responded that they did not know whether the major ones. Hill (1994) reported that new
thrips attack other crops rather than onions. However, agricultural practices, such as the use of improved
20% of the respondents answered that they observed varieties and use of nitrogenous fertilizer, are
thrips on garlic similar to onion. According to the responsible for changing plant hoppers from minor
interviewed farmers, thrips are difficult to manage, pests to the major pest status.
highly distributed in all onion fields and became very
important pests of onion and garlic. Effect of weather factors on onion thrips
population
The problem became intensive with onion fields and Thrips population was lower during the early crop
increased from time to time according to all the stages with lower average temperature and higher
respondents. Most of the farmers knew onion thrips relative humidity. However, number of thrips
before eight years and about 27.7% of the farmers increased gradually with increasing temperature and
knew thrips only five years ago. This may not mean decreasing relative humidity (Table 2).
that thrips were not present on their fields before five
Table 2. Meteorological data during the growing period and the mean thrips population
Date of observation Mean onion Average temperature Average rain fall Relative
thrips/plant (0C) (mm) humidity (%)
04 January 2016 15.63 25.5 00 68
22 January 2016 23.18 26.0 00 59
07 February 2016 46.20 27.5 00 36
22 February 2016 92.6 29.0 00 29
07 March 2016 127.73 30.0 00 25
22 March 2016 165.50 29.0 00 27
07 April 2016 51.97 28 00 46
Source: Ethiopian Meteorological Agency Mekelle branch and Aksum airport.
Figure 2. Mean number of thrips population /plant observed at different dates in onion

y=-742.29x+29.3 y=192.49x-2.84
r= 0.937** r= -0.929**
Figure 3. Relationship between average temperatures Figure 4. Relationship between relative humidity
(C) and thrips population in onion (%) and thrips population in onion
Table 3. Coordinates and elevations of the surveyed areas

Area North East Altitude (m.a.s.l.)
Mereb-leke 1422.824' 3848.278' 1407
1422.916' 3848.097' 1386
1422.828' 3848.318' 1415
1422.847' 3848.298' 1404
1423.721' 3848.272' 1370
1423.665' 3848.317' 1368
1423.649' 3848.291' 1387
1423.736' 3848.244' 1379
1423.684' 3848.291' 1350
1423.652' 3848.248' 1381
Ahferom 1411.144' 3904.256' 1995
1411.219' 3904.197' 1999
1416.374' 3903.934' 2020
1411.096' 3904.225' 2007
1411.129' 3904.289' 2007
1411.144' 3904.030' 2004
1416.390' 3930.936' 2029
1416.379' 3904.330' 2006
1416.359' 3904.320' 2014
1411.145' 3904.248' 2005
Adwa 1409.494' 3851.247' 1894
1409.497' 3851.251' 1897
1409.101' 3851.126' 1879
1409.133' 3851.096' 1876
1408.593' 3852.289' 1830
1408792' 3852.645' 1841
1408.742' 3852.547' 1837
1408.318' 3852.396' 1835
1408.553' 3852.262' 1833
1408.667' 3852327' 1841
Laelay-mychew 1407.189' 3845.809' 2088
1407.265' 3845.865' 2089
1407.293' 3845.885' 2082
1407.379' 3845.813' 2086
1407.050' 3837.182' 2131
1407.910' 3846.147' 2093
1407.998' 3846.087' 2102
1407.784' 3846.024' 2098

There was strongly positive correlation between others observed 30-50% yield reduction when control
temperature and thrips population in onion. The measures are not taken 27.5 and 50% of the
regression equation was y = -742.29x+29.33 and respondents, respectively. Most of the onion growers
correlation coefficient was r = 0.937**. Figure (3) in the surveyed areas used insecticides only,
indicated that thrips population was increased with particularly lambda-cyhalothrin (Karate) for onion
the increase of temperature. However, Thrips thrips management even though it was not effective
population was negatively correlated with relative and expensive to purchase. Most onion fields need an
humidity in onion. The regression equation was y = integrated management against onion thrips for two to
192.49x-2.84 and correlation coefficient was r = - three months depends on the weather conditions.
0.929**. Figure (4) indicated that thrips population
decreased with the increase of relative humidity. Acknowledgement:
The research work reported in this paper is in
Similar findings have been reported by different collaboration with my home institution Tigray
authors on climatic factors, including the important Agricultural research Institute, Aksum Agricultural
factors rainfall, temperature, relative humidity and research Center. I am very grateful to, Tigray
wind that significantly affect thrips numbers (Kirk, Agricultural Research Institute for the financial
1997; Waiganjo et al., 2008). Relatively high support and Akum University for providing me with
temperature and lack of rainfall increased onion thrips laboratory equipment for pest identification. I would
population, while high relative humidity and rainfall like to express my thanks and appreciation to Mulatu
reduced thrips population (Hamdy and Salem, 1994; wakgari (PhD), Gashawbeza Ayalew (PhD), Mashila
Tadele and Amin, 2014). Effect of weather factors on Dejjene (PhD), Tesfay Belay (PhD) and Firdu
thrips colonization depicted that heavy rainfall and Azerefegn (PhD) for their critical review of the paper.
low temperature had negative correlation with thrips It has helped me to improve the quality of the report
abundance (Haider et al., 2014). Similarly, Lorini and and compilation of my work.
Junior (1990) reported that high temperatures and
lack of rainfall increased density of T. tabaci on garlic Conflict of interest: The author stated that no
in Brazil. conflicts of interest.
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Abate (eds). Pages 61-67.
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RESEARCH ARTICLE
The Genetic and Morphological Evolution of Fish through the

Protein Cytochrome C Oxidase Subunit One
Sammer Marzouk,

Received: 12.07.2017 Background: Cytochrome C Oxidase Subunit 1 (COX1) is a protein that

Accepted: 14.08.2017 helps to catalyze the reduction of water into oxygen in Eukaryotes. Through
Published : 19.09.2017 the analyzation of COX1 from online public genetic databases in 16 species
of fish, an evolutionary phylogeny of fish was derived from the data. This
Editor: paper considered three hypotheses: Axolotl (Ambystoma mexicanum) and
Dr. Arvind Chavhan Tiktaalik (Tiktaalik roseae) share a common ancestor was determined; that
A.gueldenstaedtii and P.spathula both share a common ancestor; and that
P.marinus and S.acanthias are the outliers of the phylogeny. The
Sammer Marzouk (2017) The
evolutionary phylogeny used the percent ID between the two species of fish.
Genetic and Morphological Evolution
of Fish through the Protein From these differences, analysis is done to the data and the data is used to
Cytochrome C Oxidase Subunit One; make phylogenies based on the morphological and genetic evolution of
International J. of Life Sciences, 5 (3): these fish.
332-344. Results: From the data derived from the phylogenies, the results
demonstrates the claims that Axolotl (Ambystoma mexicanum) and Tiktaalik
(Tiktaalik roseae) share a common ancestor was determined,
Copyright: 2017| Author (s), This A.gueldenstaedtii and P.spathula both share a common ancestor, and
P.marinus and S.acanthias are the outliers of the phylogeny.
Conclusion: The data gathered can be used to connect tetrapods to fish,
Derivs License, which permits use and contribute to the theory that tetrapods came from fish. The information
and distribution in any medium, presented in this paper can be used to make a complete phylogeny of all
provided the original work is organisms in the biosphere.
Keywords: Cladogram, Evolutionary Biology, Ichthyology, Phylogeny, and
adaptations are made. Evolution.
INTRODUCTION
Of the 30,000 species of fish, the phylogenetic trees of fish have not been
heavily studied on built upon for the reason that their has not been a
reliable connection between fish and tetrapods (Fiz-Palacios et al., 2013).
Charles Darwin first proposed that animals could be categorized by the
morphological traits they possessed, and supported this by categorizing
finches based on their beaks. This type of phylogeny made by data from
morphological traits is the morphological phylogeny. As DNA sequencing
technology because in greater use, a new type of phylogeny from which

The Genetic and Morphological Evolution of Fish through the Protein Cytochrome C Oxidase
protein-sequences were used was also used to Table 1: Sample group of 16 species of fishes
categorize animals. These two types of phylogenies
Common Name: Genus Species:
would show a hypothesis of the evolution of certain
beings. But sometimes there would be differences in Spiny Dogfish Squalus acanthias
these two types of phylogenies. This demonstrates the Sea Lamprey Petromyzon marinus
need of how can a cladogram be inferred using genetic
Saddled Bichir Polypterus endlicheri
data be reconciled with a cladogram using only
morphological data? From a sample size of 16 species Paddlefish Polyodon spathula
of fish, a morphological phylogeny was previously Ghost Shark Callorhynchus milii
derived (Fig.1). The phylogeny has 21 traits and shows Coelacanth Latimeria chalumnae
which traits evolved in which species (Fig.1). From the
morphological phylogeny, it is shown that gills, Bowfin Amia calva
spiracles, and a cartilage skeleton are the basic traits Pollen`s Cichlid Paratilapia pollen
that make this clade of species (Fig.1). Overtime, more Tiktaalik Tiktaalik roseae
diverse and complex traits divided up the fish into
Ropefish Erpetoichthys calabaricus
separate taxa (Fig.1). In the morphological phylogeny
Tiktaalik (Tiktaalik roseae), and Axolotl (Ambystoma Great White Shark Carcharodon carcharias
mexicanum) were both in the same clade, which lead to Axolotl Ambystoma mexicanum
the hypothesis that if the morphological and protein
Cuvier's Bichir Polypterus senegalus
phylogenies were compared, then it would determine
that they shared a common ancestor (Fig.1). Also, in Russian Sturgeon Acipenser gueldenstaedtii
the morphological phylogeny, it is shown that Australian Lungfish Neoceratodus forsteri
A.gueldenstaedtii and P.spathula both share a common
Goldfish Carassius auratus
ancestor, and P.marinus and S.acanthias are the
outliers of the phylogeny (Fig. 1). In order to get the
most accurate results, this morphological data was
combined with the protein-sequence of Cytochrome C
Oxidase Subunit 1 (COX1) (Table 2). Cytochrome C phylogeny was compared with the morphological
Oxidase Subunit 1 (COX1) is one out of the three phylogeny to make a final phylogeny. This paper
mitochondrial DNA subunits, the others being MT-CO2, describes the morphological and protein similarity
MT-CO3 that are part of respiratory complex IV between the 16 species of fish, and describes the use of
(Tsukihara et al., 1996). In respiratory complex IV, this building a more extensive phylogenetic tree.
enzyme completes the electron transport system, and
its function of catalyzing the reduction of water into
oxygen (Tsukihara et al., 1996). Most Eukaryotes, and MATERIALS AND METHODS
all vertebrates have this protein (Tsukihara et al.,
1996). The COX1 from the Spiny Dogfish (Squalus Morphological Phylogeny
acanthias), Sea Lamprey (Petromyzon marinus), The morphological tree phylogeny was constructed
Saddled Bichir (Polypterus endlicheri), Paddlefish based off of 21 synapomorphies of a sample group of
(Polyodon spathula), Ghost Shark (Callorhynchus milii), 16 species of fish (Fig. 1). A species would be
Coelacanth (Latimeria chalumnae), Bowfin (Amia compared with the other species of fish. If the species
calva), Pollen`s Cichlid (Paratilapia pollen), Tiktaalik had or did not have a specific trait, they would be
(Tiktaalik roseae), Ropefish (Erpetoichthys placed in a different clade than species that did not did
calabaricus), Great White Shark (Carcharodon not have another trait (Fig. 1). The continuation of this
carcharias), Axolotl (Ambystoma mexicanum), Cuvier's pattern would give a complete morphological
Bichir (Polypterus senegalus), Russian Sturgeon phylogeny (Fig. 1). The morphological phylogeny was
(Acipenser gueldenstaedtii), Australian Lungfish based off of a data matrix which was derived from the
(Neoceratodus forsteri), and Goldfish (Carassius characteristics that the species of fish might or might
auratus) were sequenced and compared in order to not have (Table 3). For every characteristic, if it was
find the percent ID (Table 1). This percent ID was used present in that species of fish it had a Y otherwise it
in order to make a phylogeny. The protein-sequenced had a N in it (Table 3).

Protein Sequences and Construction of Phylogeny deriving a spot for the species based on a specialized
The protein sequence of the fish was derived from an computer algorithm. Any contradictions would be
online public genomic database (Table 4). The data resolved by analysing the percent similar and
was then compared and analyzed by a series of determining which species had the greater percent
specialized computer algorithms. These computer similar (Fig. 3). This species would then be placed
algorithms gave the specific percent identity (specific accordingly (Fig. 3).
as in rounded to the nearest tenth) and the specific
percent similar. These percentages were then used, 2.4 Data Analysis
and compiled on a genetic chart. From the genetic All sequenced protein data has been verified by the
chart, the phylogenetic tree was derived (Table 4). In proper scientific body. Data analysis was by a
order to make the phylogeny, the data on the first specialized computer algorithm developed by William
species, S.acanthias was compared to the rest of the Pearson and run on the LALIGN server. All data was
fish (Fig. 2). The one with the largest percent identity tested for specific outliers and the percent similar
was put into a clade with S.acanthias (Fig. 2). After induced function of amino acids. percent identity
this, the next species that was closest to S.acanthias included exact amino acid comparisons. Identical and
was put in the same clade the two previous fish. similar percentages were rounded to the nearest tenth.
Combination of Morphological and Protein

Phylogeny RESULTS
In order to combine the types of phylogenies, further
examination on the similarities and differences of the From the data, three specific conclusions were drawn.
phylogenies would be conducted (Fig. 3). Similarities The first conclusion drawn from the data was that
would be kept and considered a constant of both Axolotl (Ambystoma mexicanum) and Tiktaalik
phylogenies (Fig. 3). Differences in species placement (Tiktaalik roseae) share a common ancestor (Fig.1,
were decided by comparing the two phylogenies and Fig.2, Fig.3).
Fig. 1 Morphological Phylogeny. The morphological phylogeny is showing the 16-different species of fish,
and their synapomorphic distinctions. From the table, it is shown the morphologically, the Sea Lamprey is
most distinct to pollens Cichlid. Not all possible traits are shown by the phylogeny.

Fig. 2 Protein-Derived Phylogeny. This is the phylogeny that was derived using the protein comparisons of
COX 1 from the 16 species of fish. The data demonstrates that the first species, the Sea Lamprey is an outlier
from the rest of the fish.
Fig. 3 Final Phylogeny. This is the final phylogeny that was made by combining the morphological phylogeny and
the protein derived phylogeny. The final phylogeny demonstrates the impformation that Tiktaalik is closely
related to pollens Cichlid.

Table 2 : Morphological data was combined with the protein-sequence of Cytochrome C Oxidase Subunit 1 (COX1) from the 16 species of fish
www.ijlsci.in
White Shark
Coelacanth
Paddlefish
Australian
Sturgeon
Lamprey
Tiktaalik
Ropefish
Lungfish
Goldfish
Cuvier's
Saddled
Pollen`s
Russian
Dogfish
Bowfin
Axolotl
Cichlid
Bichir
Bichir
Ghost
Shark
Great
Sea
Sea
Sea 87.3/ 90.4/ 92.4/ 90.9/ 89.5/ 90.7/ 92.2/ 91.6/ 94.2/ 89.3/ 90.8/ 92.4/ 91.9/ 93/
100%
Dogfish 97.9 98 98.4 97.7 97.7 98.3 98.6 98.4 98.5 97.9 98.6 98.6 98.6 98.6
Sea 87.3/ 87.9/ 88.9/ 87.6/ 89.1/ 88.3/ 88.1/ 87.9/ 86.3/ 86.2/ 88.3/ 89.1/ 86.9/ 88.9/
100%
Lamprey 97.9 96.5 97.5 96.3 97.3 97.3 97.1 96.9 97. 96.5 96.9 97.9 97.7 97.5
Saddled 90.4/ 87.9/ 92.4/ 90.6/ 91.5/ 91.7/ 92.6/ 98.5/ 90.4/ 90.3/ 98.6/ 92.6/ 91.6/ 92.2/
100%
Bichir 98 96.5 98.6 98 98.3 98.6 98.3 99.6 98.2 97.7 99.4 99.2 99 98.8
92.4/ 88.9/ 92.4/ 90.5/ 93.8/ 95.1/ 95.3/ 93.4/ 91.2/ 89.1/ 90.8/ 98.5/ 93.2/ 95.9/
Paddlefish 100%
Int. J. of Life Sciences, Vol. 5(3) September 2017
98.4 97.5 98.6 98.6 99 99.4 99.6 99 98.1 97.5 98.2 99.8 99.6 99.4
Ghost 90.9/ 87.6/ 90.6/ 90.5/ 91/ 90/ 91.1/ 90.6/ 91.2/ 89.1/ 90.8/ 90.9/ 91.1/ 90.1/
100%
Shark 97.7 96.3 98 98.6 98.2 98.4 98.4 98.2 98.1 97.5 98.2 98.8 98.8 98.3
89.5/ 89.1/ 91.5/ 93.8/ 91/ 92.2/ 93.4/ 92/ 89.5/ 90.8/ 92/ 93.8/ 93.8/ 92.8/
Coelacanth 100%
97.7 97.3 98.3 99 98.2 98.8 99 98.2 97.9 98.6 98.4 99.2 98.8 98.8
90.7/ 88.3/ 91.7/ 95.1/ 90/ 92.2/ 93.6/ 93/ 89.9/ 91.6/ 92.4/ 95.1/ 92.4/ 93.4/
Bowfin 100%
98.3 97.3 98.6 99.4 98.4 98.8 99.2 99.2 98.3 98.4 98.8 99.6 99.2 99
Pollen`s 92.2/ 88.1/ 92.6/ 95.3/ 91.1/ 93.4/ 93.6/ 93.9/ 92.2/ 93.6/ 93.8/ 95.9/ 94.7/ 94.9/
100%
Cichlid 98.6 97.1 98.3 99.6 98.4 99 99.2 99 99 98.6 98.8 99.8 99.4 994
Tiktaalik 100%
91.6/ 87.9/ 98.5/ 93.4/ 90.6/ 92/ 93/ 93.9/ 91/ 90.8/ 98.6/ 95.9/ 91.5/ 92/
Ropefish 100%
98.4 96.9 99.6 99 98.2 98.2 99.2 99 98.6 98.2 99.8 99.6 99 98.8
Great
94.2/ 86.3/ 90.4/ 91.2/ 91.2/ 89.5/ 89.9/ 92.2/ 91/ 89.3/ 90.6/ 91.9/ 92.2/ 91.8/
White 100%
98.5 97.1 98.2 99 98.1 97.9 98.3 99 98.6 97.9 98.8 98.6 99 99
Shark
89.3/ 86.2/ 90.3/ 92.4/ 89.1/ 90.8/ 91.6/ 93.6/ 90.8/ 89.3/ 91.4/ 92.8/ 91.2/ 91.8/
Axolotl 100%
97.9 96.5 97.7 98.9 97.5 98.6 98.4 98.6 98.2 97.9 98.4 99 98.6 99
Cuvier's 90.8/ 88.3/ 98.6/ 93.2/ 90.8/ 92/ 92.4/ 93.8/ 98.6/ 90.6/ 91.4/ 92.8/ 93/ 93/
100%
Bichir 98.6 96.9 99.4 99.2 98.2 98.4 98.8 98.8 99.8 98.8 98.4 99 99.6 99.4
Russian 92.4/ 89.1/ 92.6/ 98.5/ 90.9/ 93.8/ 95.1/ 95.9/ 93.6/ 91.9/ 92.8/ 95.9/ 94.2/ 95.9/
100%
Sturgeon 98.6 97.9 99.2 99.8 98.8 99.2 99.6 99.8 99.6 98.6 99 99.6 99.8 99.6
Australian 91.9/ 86.9/ 91.6/ 93.2/ 91.1/ 93.8/ 92.4/ 94.7/ 91.5/ 92.2/ 91.2/ 93/ 94.2/ 93/
100%
Lungfish 98.6 97.7 99 99.6 98.8 98.8 99.2 99.4 99 99 98.6 99.6 99.8 99.6
93/ 88.9/ 92.2/ 95.9/ 90.1/ 92.8/ 93.4/ 94.9/ 93.2/ 92/ 91.8/ 93/ 95.9/ 93/
Goldfish 100%
98.6 97.5 98.8 99.4 98.3 98.8 99 994 99.2 98.8 99 99.4 99.6 99.6
336

Table 3 : The morphological characteristics that the species of fish might or might not have
Feature
Coelacanth
Paddlefish
Lamprey
Tiktaalik
Ropefish
Saddled
Pollens
Dogfish
Bowfin
Cichlid
Bichir
Shark
Ghost
Spiny
Sea
gills Y Y Y Y Y Y Y Y Y Y
spiracles Y Y Y Y Y Y Y Y Y Y
Cartilage skeleton Y Y Y Y Y Y Y Y Y Y
jaws Y N Y Y Y Y Y Y Y Y
Paired appendages Y N Y Y Y Y Y Y Y Y
Placoid scales Y N Y Y Y Y Y Y Y Y
Jelly filled swim
N N Y N N Y Y Y Y Y
bladder
Scales reduced N N Y N N Y Y Y Y Y
Air filled swim
N N Y N N Y Y Y Y Y
bladder
Ossification of
N N Y N N Y Y Y Y Y
skeleton
Cycloid skeleton N N Y N N Y Y Y Y Y
Swimbladder used as
N N N N N Y N N Y N
lung
Cosmoid scales N N N N N Y N N Y N
Ganoid scales N N Y N N N N N N Y
Lobed fins N N Y N N Y Y Y N Y
Dorsoventral
N N Y N N N N Y N Y
flattening
Pectoral girdle
N N Y N N N N Y N Y
extension
elasmobranch Y N N N N N N N N N
Tetrapod limbs N N N N N N N N Y N
Pectoral walking N N Y N N N N N N N
Morphologically, A.mexicanum, and T.roseae share the paired appendages, placoid scales, gills, spiracles, and
synapomorphies of tetrapod limbs, lobed fins, cosmoid cartilage skeletons (Fig. 1). And in the protein-derived
scales, an air-filled swim bladder, an ossified skeleton, phylogeny, it is shown that A.gueldenstaedtii and
cycloid scales, a reduced amount of scales, jaws, paired P.spathula have a 98.5 percent identity with each
appendages, and having a cartilage skeleton (Fig. 1). other.(Fig. 2). In the final phylogeny, it was shown that
Since T.roseae is an extinct species, no protein A.gueldenstaedtii and P.spathula with the 98.5 percent
sequence of COX1 from T.roseae was available (Fig. 2). identity and the similar synapomorphies are related
Only the morphological phylogeny could be used to and have once shared a common ancestor (Table 2).
infer where T.roseae goes on the final phylogeny (Fig.
1). In the final phylogeny, when the data from the The third conclusion derived from the data is that
previous phylogenies were combined, it was shown P.marinus and S.acanthias are the outliers of the
that A.mexicanum and T.roseae, both from protein phylogeny (Fig.1). In the morphological phylogeny,
comparisons and morphological traits, shared a P.marinus and S.acanthias were shown to share many
common ancestor (Fig.1, Fig.2, Fig.3). similar synapomorphies (Fig. 1). These include gills,
spiracles, and cartilage skeletons (Fig. 1). But in the
In the morphological phylogeny, it is shown that protein-derived phylogeny, it was demonstrated that
A.gueldenstaedtii and P.spathula are in the same clade the COX1 comparison between the two species of fish
as they both have jelly-filled swim bladders, jaws, yield an 87.3 percent identity (Table 2). This is a high

average, but compared to the other relationships, such (Ambystoma mexicanum) and Tiktaalik (Tiktaalik
as S.acanthias and A.gueldenstaedtii sharing a 90.4 roseae) share a common ancestor was derived from
percent identity, and P.marinus and P.pollen, who the morphological phylogeny (Fig. 1). In the
morphologically are different, only sharing three morphological phylogeny, both A.mexicanum and
synapomorphies by having a 90.8 percent identity T.roseae formed a clade, and shared 15
(Fig.1, Fig.5). In the final phylogeny, it was decided synapomorphies (Fig. 1). In the protein-derived
that S.acanthias and A.gueldenstaedtii would be the phylogeny, there was no evidence of a relationship
outliers of the group, because the protein-derived data between A.mexicanum and T.roseae as T.roseae is
was more accurately studied, and the synapomorphies extinct (Fig. 2). A.mexicanum and T.roseae have been
could be modified in a way to change the position of shown to share many synapomorphies (Ericsson et al.,
any of the two fish (Fig. 3). These three conclusions 2012). A.mexicanum and T.roseae both share a specific
were the three main results. Another finding revealed organization of neck muscles, which goes around their
that all of the 16 fish species are all related (Table 2). gills and allows them to move the head (Ericsson et
Of the 15 COX1 protein sequences that were available, al., 2012). A.mexicanum and T.roseae also both share
all of them had an percent identity greater than 88% appendicular skeleton which connect to their neck
(Table 2). Morphologically, the species of fish are muscles (Ericsson et al., 2012). A.mexicanum and
shown to all have similar traits, except P.marinus, T.roseae have both been shown to be closely related as
which is shown to be the outlier of the group (Table 2). they have been put on the same clade on
Tis information correlates to the protein-derived morphological phylogenies (Shubin et al., 2014).
phylogeny in which P.marinus is also the outlier of the A.mexicanum and T.roseae share a unique pelvic girdle
clade, and also in the final phylogeny in which and fin that allows for movement, and it is
P.marinus is the outlier of the clade (Fig.1, Fig.2, Fig.3). demonstrated that T.rosales pelvic girdle was a
S.acanthias is also shown to be constant in its position transitional phase for A.mexicanums pelvic girdle
in all three phylogenies. In the morphological, protein- (Shubin et al., 2014). The second conclusion was that
derived, and final phylogenies, S.acanthias was shown A.gueldenstaedtii and P.spathula both share a common
to always be in the same clade as C.carcharias (Fig.1, ancestor, which was derived from the morphological
Fig.2, Fig.3). Every species on the morphological and and protein-based phylogeny (Fig.1, Fig. 2). In the
protein-derived phylogeny were in the same position morphological phylogeny, A.gueldenstaedtii and
in the final phylogeny except P.senegalus, P. Endlicheri P.spathula were in the same clade and shared 8
and A.calva (Fig.1, Fig.2, Fig.3). In protein-derived synapomorphies (Fig. 1). In the protein-derived
phylogeny, it was demonstrated that P. Endlicheri was phylogeny, A.gueldenstaedtii and P.spathula shared a
similar to P.senegalus while in the morphological 98.5 percent identity (Table 2). Morphologically, it is
phylogeny, it was demonstrated that C.milii and A.calva supported that A.gueldenstaedtii and P.spathula are
were more related P.senegalus then P. Endlicheri was related as they both have eggs with funnels and clear
(Fig.1, Fig.2, Fig.3). groves (Ludwig, 2008). Also, A.gueldenstaedtii and
P.spathula share the same sized micropylar opening
(Ludwig, A, 2008). Genetically the data is supported as
DISCUSSION A.gueldenstaedtii and P.spathula have similar
heteroplasmic movements in the mtDNA control
In an effort to build a phylogeny that demonstrates a region (Wang et al., 2010). This allows both species to
link between fish and tetrapods, COX1 from 16 have lengths of central units 78-83 bp (Wang et al.,
different species was sequenced and made into a 2010). The conclusion that A.gueldenstaedtii and
protein-derived phylogeny (Fig. 3). This phylogeny P.spathula are related shown through the fact that
was combined with a morphological phylogeny with A.gueldenstaedtii and P.spathula are in the same
21 synapomorphies of the 16 species of fish to make a Kingdom, Phylum, Superclass, Class, and order (Carroll
final phylogeny (Fig. 1). From this final phylogeny, R, 1998). This demonstrates a support for the claim
three main conclusion were derived: A.mexicanum and that A.gueldenstaedtii and P.spathula are closely
T.roseae share a common ancestor; A.gueldenstaedtii related (Carroll, 1998). The third conclusion was that
and P.spathula both share a common ancestor; and P.marinus and S.acanthias are the outliers of the
P.marinus and S.acanthias are the outliers of the phylogeny, which was derived from the morphological
phylogeny. The first conclusion which was Axolotl and protein-derived phylogeny (Fig. 2, Fig. 3). In the

morphological phylogeny, P.marinus and S.acanthias been shown that P.marinus and S.acanthias have
share three synapomorphies, and were in different different gas transfer rates due to the difference in
clades (Fig. 1). In the protein-derived phylogeny, respiratory system IV (Randall, 1995). And it has been
P.marinus and S.acanthias share an 87.3 percent demonstrated that P.marinus and S.acanthias are both
identity (Fig. 2). This is the lowest percent identity of in genetically separate groups, which share a 0.400
the 15 species of fish that had a COX1 protein similarly plasma level rate (Steve et al., 1998).
sequence available (Table 2). It is supported that P.marinus and S.acanthias are also shown to be put
P.marinus and S.acanthias are morphologically into separate clades (Grim et al., 2011). P.marinus and
different, as P.marinus and S.acanthias have different S.acanthias are demonstrated to be not very related, as
secretory granule compositions (Wilson et al., 2002). they are in different classes, subclasses, orders,
P.marinus and S.acanthias also have a different type of families and genuses (Grim et al., 2011). This finding
microfilament network (Wilson et al., 2002). that P.marinus and S.acanthias shown to be a support
Genetically, the claim that P.marinus and S.acanthias for the idea that P.marinus and S.acanthias are not very
are outliers of the fish phylogeny is supported. It has related (Grimm et al., 2011).
Table 4: The protein sequence of the fish was derived from an online public genomic database
Spiny Dogfish:
>sp|Q9ZZ52|COX1_SQUAC Cytochrome c oxidase subunit 1 OS=Squalus acanthias GN=MT-CO1 PE=3 SV=1
MAINRWFFSTNHKDIGTLYLIFGAWAGMVGTALSLLIRAELSQPGTLLGDDQIYNVIVTA
HAFVMIFFMVMPVMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSLLLLLASAGVEA
GAGTGWTVYPPLAGNMAHAGASVDLAIFSLHLAGISSILASINFITTIINMKPPAISQYQ
TPLFVWSILVTTILLLLSLPVLAAAITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMISHVVAYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGSIKWETPLLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMAGFIHWFPLFSGYTLHSTWTKTQFLVMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYALWNTVSSIGSLISLVAVIMFLFIIWEAFAPSG
KVLSVELPHTNVEWLHGCPPPYHTYEEPAFVQVQRTYF
Sea Lamprey
>sp|Q35536|COX1_PETMA Cytochrome c oxidase subunit 1 OS=Petromyzon marinus GN=MT-CO1 PE=3 SV=1
MTHIRWLFSTNHKDIGTLYLIFGAWAGMVGTALSILIRAELSQPGTLLGDDQIFNVIVTA
HAFVMIFFMVMPIMIGGFGNWLVPLMLGAPDMAFPRMNNMSFWLLPPSLLLLLASAGVEA
GAGTGWTVYPPLAGNLAHTGASVDLTIFSLHLAGVSSILGAVNFITTIFNMKPPTMTQYQ
TPLFVWSVLITAVLLLLSLPVLAAAITMLLTDRNLNTSFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGIISHVVAYYAGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGKIVWHTPMLWALGFIFLFTVGGLTGIVLSN
SSLDIILHDTYYVVAHFHYVLSMGAVFAIMAGFVHWFPLFTGYTLNETWAKAHFIIMFAG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTTWNIISSIGSTVSLIAVMLFMFILWEAFSAKR
KAIATDLLNTNLEWLHGCPPPYHTYEEPAFVQTNFKK
Saddled Bichir:
>tr|F2W4F6|F2W4F6_9ACTI Cytochrome c oxidase subunit 1 OS=Polypterus endlicherii congicus GN=COX1 PE=3
SV=1
MTITRWLFSTNHKDIGTLYLIFGAWAGMVGTALSLLIRAELGQPGALMGDDQIYNVIVTA
HAFVMIFFMVMPIMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSLLLLLTSSAVEA
GVGTGWTVYPPLAGNLAHAGASVDLAIFSLHLAGVSSILGAINFITTIINMKSPSTSQYQ
TPLFVWSVLVTAILLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMISHIVAYYSGKNEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGAIKWETPMLWALGFIFLFTVGGLTGIILAN
SSLDIMLHDTYYVVAHFHYVLSMGAVFAIMGGFVHWFPLFSGYTLHPTWTKIHFGIMFIG

VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNSLSSIGSMISLTAVIMFLFILWEAFAAKR
EVQMVNLTYTNVEWLHGCPPPYHTYEEPAFVQSPQARE
Paddlefish:
>tr|Q8HMQ8|Q8HMQ8_POLSP Cytochrome c oxidase subunit 1 OS=Polyodon spathula GN=COI PE=3 SV=1
MAITRWFFSTNHKDIGTLYLVFGAWAGMVGTALSLLIRAELSQPGALLGDDQIYNVIVTA
HAFVMIFFMVMPIMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASSGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGVSSILGAINFITTIINMKPPAVFQYQ
TPLFVWSVLVTAVLLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMISHIVAYYAGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGSIKWDTPLLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMGAFVHWFPLFTGYTLHSTWSKIHFAVMFVG
VNLTFFPQHFLGLAGMPRRYSDYPDAYALWNTVSSVGSLISLIAVIMFLFILWEAFAAKR
EVLSVELTATNVEWLHGCPPPYHTYEEPAFVQVQSTN
Ghost Shark:
>tr|D7RWS9|D7RWS9_CALMI Cytochrome c oxidase subunit 1 OS=Callorhinchus milii GN=COX1 PE=3 SV=1
MTINRWLFSTNHKDIGTLYLLFGAWAGMVGTALSLLIRAELSQPGALMGDDQIYNVIVTA
HAFVMIFFMVMPIMIGGFGNWLIPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASAGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGISSILASINFITTIINMKPPSITQYQ
TPLFVWSILITTILLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMISHVVTYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGNIKWDTPMLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMAGLVHWFPLFTGYTLHETWTKIHFGLMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNSVSSIGSLMSLLAVILFLFILWEAFASKR
TLSHVMMSSTNNEWLHGCPPPHHTFEEPAFVQIQVNKKG
Ghost Shark:
>tr|D7RWS9|D7RWS9_CALMI Cytochrome c oxidase subunit 1 OS=Callorhinchus milii GN=COX1 PE=3 SV=1
MTINRWLFSTNHKDIGTLYLLFGAWAGMVGTALSLLIRAELSQPGALMGDDQIYNVIVTA
HAFVMIFFMVMPIMIGGFGNWLIPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASAGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGISSILASINFITTIINMKPPSITQYQ
TPLFVWSILITTILLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMISHVVTYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGNIKWDTPMLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMAGLVHWFPLFTGYTLHETWTKIHFGLMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNSVSSIGSLMSLLAVILFLFILWEAFASKR
TLSHVMMSSTNNEWLHGCPPPHHTFEEPAFVQIQVNKKG
Coelacanth:
>sp|O03167|COX1_LATCH Cytochrome c oxidase subunit 1 OS=Latimeria chalumnae GN=MT-CO1 PE=3 SV=1
MMITRWLFSTNHKDIGTLYMIFGAWAGMVGTALSLLIRAELSQPGALLGDDQIYNVVVTA
HAFVMIFFMVMPIMIGGFGNWLIPLMIGAPDMAFPRMNNMSFWLLPPSLLLLLASSGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGVSSILGAINFITTVINMKPPTMTQYQ
TPLFIWSVLVTAVLLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMISHIVAYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGVTKWDTPLLWALGFIFLFTVGGLTGIVLAN
SSLDIILHDTYYVVAHFHYVLSMGAVFAIMGGLVHWFPLMTGYTLHNTWTKIHFGVMFTG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNTVSSIGSLISLIAVIMFMFILWEAFSAKR
EVLIVEMTTTNVEWLHGCPPPHHTYEEPAFVQAPR

Bowfin:
>tr|Q8HQL9|Q8HQL9_AMICA Cytochrome c oxidase subunit 1 OS=Amia calva GN=COI PE=3 SV=1
MTITRWFFSTNHKDIGTLYLVFGAWAGMVGTALSLLIRAELSQPGALLGDDQIYNVIVTA
HAFVMIFFMVMPVMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASSGVEA
GAGTGWTVYPPLASNLAHAGASVDLTIFSLHLAGVSSILGAINFITTIINMKPPAASQYQ
TPLFVWSVLITAVLLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMVSHIVAYYAGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMVIAIPTGVKVFSWLATLHGGAIKWETPLLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMGGFVHWFPLFSGYTLHPTWSKIHFGVMFVG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNTVSSIGSLVSLIAVVMFLFMLWEAFAAKR
EVLAVEYAATNVEWLHGCPPPYHTYEEPAYVQAHRA
Pollen`s Cichlid:
>tr|B5DCF6|B5DCF6_9CICH Cytochrome c oxidase subunit 1 OS=Paratilapia polleni GN=CO1 PE=3 SV=1
MAITRWFFSTNHKDIGTLYLVFGAWAGMVGTALSLLIRAELSQPGSLLGDDQIYNVIVTA
HAFVMIFFMVMPIMIGGFGNWLIPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASSGVEA
GAGTGWTVYPPLAGNLAHAGPSVDLTIFSLHLAGVSSILGAINFITTIINMKPPAISQYQ
PEVYILILPGFGMISHIVAYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGAIKWDTPLLWALGFIFLFTVGGLTGIVLAN
SSLDIMLHDTYYVVAHFHYVLSMGAVFAIVGGFVHWFPLFSGYTLHSTWTKIHFGVMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNTVSSIGSMISLVAVIMFLFIIWEAFAAKR
EVLSAELTTTNAEWLHGCPPPYHTFEEPAFVQVQQA
Ropefish:
>tr|Q8HMU1|Q8HMU1_ERPCA Cytochrome c oxidase subunit 1 OS=Erpetoichthys calabaricus GN=COI PE=3 SV=1
HAFVMIFFMVMPIMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLTSSAVEA
GVGTGWTVYPPLAGNLAHAGASVDLAIFSLHLAGVSSILGAINFITTIINMKPPATSQYQ
SSLDIMLHDTYYVVAHFHYVLSMGAVFAIMGGFVHWFPLFSGYTLHPTWTKIHFGVMFIG
EVQTVNLTYTNVEWLHGCPPPYHTYEEPAFVQSPNSRE
Great White Shark:

>tr|I1U3D7|I1U3D7_CARCH Cytochrome c oxidase subunit 1 OS=Carcharodon carcharias PE=3 SV=1
MAINRWLFSTNHKDIGTLYLIFGAWAGMVGTALSLLIRAELGQPGSLLGDDQIYNVIVTA
HAFVMIFFMVMPIMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASAGVEA
GAGTGWTVYPPLASNLAHAGPSVDLAIFSLHLAGISSILASINFITTIINMKPPAISQYQ
TPLFVWSILVTTILLLPALPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGFGMISHVVAYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGTIKWDTPLLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMAGFIHWFPLMSGFTLHQTWTKIQFTVMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNAISSIGSLISLVAVIMLLFIIWEAFASKR
EVLSIELPHTNVEWLHGCPPPYHTYEEPAFVQIQRPSF

Axolotl:
>tr|Q70EE6|Q70EE6_AMBME Cytochrome c oxidase subunit 1 OS=Ambystoma mexicanum GN=COI PE=3 SV=1
MMITRWLFSTNHKDIGTLYLVFGAWAGMVGTALSLLIRAELSQPGALLGDDQIYNVIVTA
HAFVMIFFMVMPVMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASSGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGVSSILGAINFITTSINMKPASMSQYQ
TPLFVWSVLITAVLLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPVLYQHLFWFFGH
PEVYILILPGFGMISHIVTYYSAKKEPFGYMGMVWAMMSIGLLGFIVWAHHMFTVDLNVD
TRAYFTSATMIIAIPTGVKVFSWLATMHGGAIKWDAAMLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMGGFVHWFPLFSGYTLHSTWSKIHFGVMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNTVSSIGSLISLVAVIMMMFIIWEAFASKR
EVLSTELTSTNIEWLHNCPPPYHTFEEPSFVQSRI
Cuvier's Bichir:
>tr|Q8HMS1|Q8HMS1_POLSE Cytochrome c oxidase subunit 1 OS=Polypterus senegalus senegalus GN=COI PE=3
SV=1
HAFVMIFFMVMPIMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSLLLLLTSSAVEA
GVGTGWTVYPPLAGNLAHAGASVDLAIFSLHLAGVSSILGAINFITTIINMKPPSTSQYQ
SSLDIMLHDTYYVVAHFHYVLSMGAVFAIMGGFVHWFPLFSGYTLHSTWTKIHFGVMFIG
EVQTVELTYTNVEWLHGCPPPYHTYEEPAFVQSPQARE
Russian Sturgeon:
>tr|C1J916|C1J916_ACIGU Cytochrome c oxidase subunit 1 OS=Acipenser gueldenstaedtii GN=cox1 PE=3 SV=1
MAITRWFFSTNHKDIGTLYLVFGAWAGMVGTALSLLIRAELSQPGALLGDDQIYNVIVTA
HAFVMIFFMVMPIMIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASSGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGVSSILGAINFITTIINMKPPAVSQYQ
PEVYILILPGFGMISHIVAYYAGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGSIKWDTPLLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMGAFVHWFPLFTGYTLHGTWSKIHFAVMFVG
VNLTFFPQHFLGLAGMPRRYSDYPDAYALWNTVSSIGSLISLVAVIMFLFILWEAFAAKR
EVMSVELTTTNVEWLHGCPPPYHTYEEPAFVQVQSTS
Australian Lungfish:
>tr|Q94XJ5|Q94XJ5_NEOFS Cytochrome c oxidase subunit 1 OS=Neoceratodus forsteri GN=COX1 PE=3 SV=1
MTITRWFFSTNHKDIGTLYMIFGAWAGMVGTALSLLIRAELSQPGALLGDDQIYNVLVTA
HAFVMIFFMVMPIMIGGFGNWLIPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASSGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGVSSILGSINFITTIINMKPPAISQYQ
TPLFIWSVMITTILLLLSLPVLAAGITMLLTDRNLNTTFFDPAGGGDPILYQHLFWFFGH
PEVYILILPGYGMISHIVAYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGSIKWETPLLWALGFIFLFTVGGLTGIVLAN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMGGFVHWFPLMTGYTLHNTWTKIHFGVMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYTLWNTVSSIGSLISLVAVIMLLFIIWEAFAAKR
EVMSIELSPTNVEWLHGCPPPHHTFEEPAFVQVQTSQR

Goldfish:
>sp|O78681|COX1_CARAU Cytochrome c oxidase subunit 1 OS=Carassius auratus GN=mt-co1 PE=3 SV=1
MAITRWFFSTNHKDIGTLYLVFGAWAGMVGTALSLLIRAELSQPGSLLGDDQIYNVIVTA
HAFVMIFFMVMPILIGGFGNWLVPLMIGAPDMAFPRMNNMSFWLLPPSFLLLLASSGVEA
GAGTGWTVYPPLAGNLAHAGASVDLTIFSLHLAGVSSILGAINFITTTINMKPPAISQYQ
PEVYILILPGFGIISHVVAYYSGKKEPFGYMGMVWAMMAIGLLGFIVWAHHMFTVGMDVD
TRAYFTSATMIIAIPTGVKVFSWLATLHGGSIKWETPMLWALGFIFLFTVGGLTGIVLSN
SSLDIVLHDTYYVVAHFHYVLSMGAVFAIMAAFVHWFPLLTGYTLHSAWTKIHFGVMFIG
VNLTFFPQHFLGLAGMPRRYSDYPDAYALWNTVSSIGSLISLVAVIMFLFILWEAFAAKR
EVLSVELTMTNVEWLHGCPPPYHTYEEPAFVQIQSN
From the phylogenies, it was shown that species that connection to be made between primates and other
were in the same genus and families were more closely animals (Nancy et al., 2002). This connection could
related morphologically and genetically (Fig. 3). It was allow for medicine to be tested on fish before the
also shown that species that were morphologically medicines use of primates (Nancy et al., 2002). A
similar were also similar in their protein structures limitation of the protein-derived phylogeny was that
(Fig. 3). A point of controversy on the topic of T.roseae was extinct and did not have any sequences of
combining the protein-derived phylogeny and the COX1 available. Because of this, T.roseae only had one
morphological phylogeny is that other versions have source for its placement on the final phylogeny, and
put P.marinus and S.acanthias closer on the final that was the morphological phylogeny. The
phylogeny as they are morphologically similar (Fig. 3). morphological phylogeny was only based off of certain
This idea of putting P.marinus and S.acanthias close to traits. If these traits were chosen in specific ways, the
each other because they have similar synapomorphies morphological phylogeny could be theoretically
disrupts the phylogeny as A.gueldenstaedtii and controlled and be made to look any way the
P.spathula are more morphologically similar and have synapomorphies allow it to be. This could have
similar COX1 sequences (Fig.5, Fig. 1). Putting affected the final phylogeny, as it allows for variations
P.marinus and S.acanthias in place of A.gueldenstaedtii of the phylogeny. The process of combining
and P.spathula would push A.gueldenstaedtii and morphological and protein-derived phylogenies is a
P.spathula apart on the phylogeny, even though they flawed process in itself (Gontcharova. A et al., 2003).
are more closely related (Fig. 3). In the phylogeny, Even though computers and mathematical formulas
there are only 16 limited taxa for the 16 species of fish make the combination more precise, phylogenies
to be put on. If one fish's position in the phylogeny made from the combination of morphological and
changes, it will impact the rest of the phylogeny. These protein-derived phylogenies suffer from inadequate,
results build upon the idea that tetrapods and fish may noncongruent taxon sampling, long-branch attraction,
share a common ancestor, and tetrapods may once or conflicting evolutionary models of the genes
been fish. This idea has implications throughout analyzed (Gontcharova et al., 2003). In summary, the
biological sciences (Nancy et al., 2002). A phylogeny present data suggests that as time progressed, fish
connecting tetrapods to fish could build a more started to gain more tetrapod-like synapomorphies,
detailed connection of the evolution of genes (Nancy et and supports the theory that tetrapods came from fish.
al., 2002). Additional studies concerning the morphological and
genetic similarity between fish and terrestrial
organisms might help better understand the link
CONCLUSION between tetrapods and fish.
This information could be more helpful as it could be

used in the medical field to find specific new nutrients Conflicts of interest: The authors stated that no conflicts of
and new antibiotics (Nancy B et al., 2002). A phylogeny interest.
between tetrapods to fish could also allow for new

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.

RESEARCH ARTICLE
Performance of bread wheat (Triticum aestivum L.) in response to

supplemental irrigation and rate of nitrogen application in
Enderta, Tigray, Northern Ethiopia
Beyenesh Zemichael1, Nigussie Dechassa2
1
Tigray Agricultural Research Institute, P.O. Box 498, Mekelle, Tigray, Ethiopia
2Haramaya University, College of Agriculture and Environmental Sciences, P. O. Box 138, Dire Dawa, Ethiopia
*Corresponding author. E-mail: beyeneshzm@yahoo.com

Received: 18.07.2017 Moisture stress and nitrogen deficiency are the most important factors
Accepted: 17.08.2017 which limit wheat production in the semi-arid Tigray region of Ethiopia.
Published : 19.09.2017 Therefore, a field experiment was conducted in Enderta district in the
region during the 2013 and 2014 cropping seasons to elucidate the
Editor:
response of bread wheat to supplemental irrigation (SI) and rates of
Dr. Arvind Chavhan
nitrogen application. The experiment was laid out as a randomized
Cite this article as: complete block design (RCBD) in a split-plot arrangement with three
Beyenesh Zemichael and Nigussie replications. Four levels of supplementary irrigation (SI), viz: 0, 50, 75, and
Dechassa (2017) Performance of 100% (full) of the crop water requirement were assigned to the main plots
bread wheat (Triticum aestivum L.)
in response to supplemental whilst, nitrogen levels of 0, 23, 46, 69 and 92 kg N ha-1 were assigned to the
irrigation and rate of nitrogen sub-plots. Application of 92 kg N ha1 with 75% SI increased tiller
application in Enderta, Tigray, production by 94% as compared to the rain-fed treatment. Similarly, plants
Northern Ethiopia; International J. of
that received 100% SI with the application of 69 kg N ha-1 had their heights
Life Sciences, 5 (3): 345-361.
significantly increased by about 39%, spike length by 56%, kernel per spike
Acknowledgements by 51%, TKW by 47% and aboveground biomass yield by 340% as
The authors gratefully acknowledge compared to no irrigation and no nitrogen application. Maximum grain
the support of East Africa Agriculture
yield of 4504kg ha-1 and water productivity of 7.99 kg mm-1 ha-1 was
Productivity Project (EAAPP) of the
Ethiopian Institute of Agricultural recorded in response to the application of 100% SI combined with 92 kg N
Research for funding this research. ha-1, However, the lowest grain yield of 885 kg ha -1 and water productivity
The authors wish to express their of 2.15 kg mm-1 ha-1 were recorded in response to the rain-fed production
appreciation also to Ato Muluberhan
Kifle and Tesfay G/Tsadik for their with no application of nitrogen fertilizer.. Based on economic analysis, the
help in the analysis of crop wat acceptable marginal rate of return of 255% and the highest net benefit
requirement of the crop (47004 ETB ha-1) was obtained from the application of 92 kg N ha -1 with
100% supplemental irrigation. It can be concluded that the integration of
is an open access article under the 92 kg N ha1 with 100% supplementary irrigation resulted in optimum
terms of the Creative Commons grain yield and economic return of the crop.
Key words: Deficit irrigation, economic analysis, N application, water
provided the original work is productivity yield, yield component

Beyenesh and Nigussie, 2017
INTRODUCTION than double rain-fed yields. Similarly, Oweis and

Hachum (2003) reported that wheat yields can be
Bread wheat (Triticum aestivum L.) one of the most increased from 2 tons per hectare to more than 5 tons
important small cereal crop which ranks fourth in per hectare by the conjunctive use and timely
terms of area coverage after teff, maize and sorghum. application of only 100 to 200 mm of irrigation water.
The crop is also one of the most important cereal crops Related studies in the country have shown that more
being cultivated in the mid and high land areas of than 80% of the yield reduction and more than 50% of
Tigray region cultivated over 100,175 hectares (ha) crop failures in tef and barley can be avoided if SI is
with a total production of about 192,507 tons (CSA, employed during the critical growth stages of the
2014). crops (Araya and Stroosnijder, 2011). SI increased the
yield of tef from less than 1 t ha-1 to 2 tons ha-1 (Yihun
Nonetheless, the productivity of wheat in the country et al., 2013 and Alemtsehay et al., 2015). Similar
is low compared to other wheat producing countries of results were also reported for Pearl millet (Ashraf et
the world (Gashaw et al., 2014). Moreover, its average al., 2001).
yield in Tigray has remained by far lower than the
national yield (Mesay et al., 2012). This explicitly However, the most beneficial effects of supplemental
shows that the production of wheat in the Region is irrigation were obtained in combination with soil
very insufficient to meet the increasing food fertility management (Rockstrm et al., 2003). Plant
demand for the ever increasing population (Gain, available nutrient supply is also essential to maximize
2014). the benefits of additional water captured or saved.
This is especially true for N (Ryan et al., 2009). Several
Though, several socio-economic, abiotic, and biotic studies suggested that limited supplemental irrigation
constraints explain these yield gaps; crop production and fertilization during the growth season could
in the semi-arid highlands of Tigray is restricted by significantly increase water productivity, fertilizer use
soil moisture stress and low fertility mainly nitrogen efficiency and wheat yield (Pandey et al., 2002;
(Fassil et al., 2009, Araya et al., 2010a). Rainfall is Mintesinot et al., 2004; Kibe et al., 2006, and Tavakoli
erratic with intermittent drought spells (Araya et al., et al., 2010). Furthermore, SI allows modifying crop
2010a), short length of growing period (Hadgu et al., calendars to escape climatic extremes and adapt to
2013). Mean annual rainfall has been estimated at 650 climate change (Oweis, 2014).
mm or less over the past few decades (Pender and
Gebremedhin, 2004), and the coefficient of variation is Supplemental irrigation (SI) is a key strategy, albeit it
four times the national level (REST 1995). However, is still underused, for unlocking rain-fed yield potential
the risk of water shortage usually happens due to late and water productivity in rain-fed wheat in Tigray
onset and early cessation of rainfall. These periods of even in areas having perennial water sources in the
water deficit mainly occur during the critical periods study area. Conversely, no research has been
of flowering, grain set and grain filling of wheat as a conducted in the study area to elucidate the effect of
result yield is consequently low(Araya et al., 2010 , application of mineral nitrogen coupled with
Alemtsehay et al., 2015). supplementary irrigation on the productivity of wheat.
The objective of the study was, therefore, to identify
Allen et al. (1998) and Rockstrm and Barron (2007) the optimum rates of nitrogen and supplemental
stated that for cereals, the period between grain filling irrigation that would maximize the productivity of
and maturity was particularly sensitive to drought bread wheat.
stress, with greatest yield reductions occurring when
stress began during or following heading or during
maturation. Water deficit around anthesis may lead to MATERIALS AND METHODS
a loss in yield by reducing spike and spikelet number
and the fertility of surviving spikelets (Giunta et al., Description of the Experimental Site
1993), while water deficit during grain-filling period A field experiment was carried out for two successive
reduces grain weight (Royo et al., 2000). seasons (2013/ 2014) under farmers field at
Enderta. The site is geographically located in the
Oweis (2014) reported that supplemental irrigation of southeast of Tigray at 130 5N latitude, 390 5 E
50200 mm in a season is sufficient to double or more longitudes and an altitude of 1970 masl. The area

Performance of bread wheat (Triticum aestivum L.) in response to supplemental irrigation
receives an average annual rainfall of 392 mm of collected in a zigzag pattern, composited, and analyzed
which 317 mm were received in the 2013 cropping at Mekelle soil laboratory research center for pH,
season. For 2014, the average annual rainfall was 790 texture, soil OC, total N, available P, CEC. for bulk
mm of which 537 mm were received in the growing density, field capacity, permanent wilting point, Soil
season. The average yearly minimum and maximum bulk density was determined from undisturbed soil
temperatures are 11 and 27 for 2013 and 11 and 26 C sample taken using a metal cylinder (core sampler) of
for 2014, respectively (Mekelle research known volume (100 cm3) that was driven into the soil
Meteorological service). The study site has a one- of desired depth and calculated as the ratio of oven dry
season rainfall pattern with extended rainy season weight of soil to a known cylinder core sampler
from March to November with the peak season in volume. The water content at field capacity was
August. The main rain season falls from June to determined in the laboratory by using a pressure
September, during which about 83% of the annual (porous) plate apparatus by applying -1/3 bar to a
rainfall occurs. The area is characterized by heavy and saturated soil sample. When water is no longer leaving
erratic rainfall distribution. the soil sample, the soil moisture was taken as field
capacity. Permanent wilting point was also determined
Experimental Materials by using pressure membrane apparatus by applying -
A bread wheat variety named Mekelle I which was 15 bars to a saturated soil sample. When water is no
released by Mekelle Agricultural Research Centre longer leaving the soil sample, the soil moisture was
(MARC) in 2011 was used as a test crop. It is a semi- taken as permanent wilting point. The improved
dwarf variety known for its moisture stress resistance recommended wheat variety known as Mekelle I was
but can perform better under good rainfall. It needs seeded on 19th July in 2013 and 12th July in 2014 by
300- 500 mm rainfall and grows at an altitude of 1980- hand drilling with a row spacing of 20cm at a seeding
2500 meters above sea level. The variety needs 90-95 rate of 150 kg ha-1.
days to reach maturity (Hintsa et al. 2011).
The full dose of P fertilizer (20 P) was applied as triple
Treatments and Experimental Design super phosphate (46% P2o5) at planting, while N was
The experiment was conducted in 2013 and 2014 split applied (half of the dose at planting and half at 30
cropping seasons. The experimental field was laid out days after emergence) as Urea: 46% N) according to
in a randomized complete block design with split plot the treatments. The improved wheat variety (Mekelle
arrangement having three replications. Four levels of I) used as test crop which was hand drilled at 150 kg
supplemental irrigation:( full irrigation or 100 %, 75 ha-1 rate with a row spacing of 20 cm was planted on
% , 50 % of the crop water requirement as well as July 19th and 12th of 2013 and 2014 cropping seasons,
rain-fed condition( control) were assigned to main respectively. The size of each plot was 3 m x 2 m
plots, while nitrogen levels (0, 23, 46, 69 and 92 kg ha- (6m2). Weeds were removed by hand weeding two
1) were maintained in subplots.. The size of each plot times at 30 and 55 days after crop emergence. The
was 3 m x 2 m (6m2) and plots and blocks were at the crop was harvested on 29th October and 2th November,
distances of 0.5 m and 1.5 m apart, respectively. Soil 2013 and 2014 respectively.
bunds were constructed around each plot and around
the entire experimental field to minimize nutrient and Determination of Irrigation Water Requirement
water movement from plot to plot. Seeds were sown The field was irrigated based on the water
into rows of 0.20 m apart and 3 m long. Each plot requirement (CWR) of the crop after the rain stopped
consisted of 10 rows. Wheat plants in the two outer (Clarke et al., 1988). To estimate the CWR, irrigation
most rows on both sides of a plot as well as 0.2 m on scheduling and irrigation water requirement (IWR) of
each end of central rows were considered border the crop the CropWat 8 for windows was used.
plants and not considered for data collection. The net Modified FAO Penman-monteith method (Allen et al.,
plot area that was used for data collection consisted of 1998) was used for calculating reference crop evapo-
6 rows (1.2 m) each 2.6 m in length. transpiration (ETO). For the computation of ETo,
meteorological data such as rainfall, Tmax, Tmin,
Experimental Procedures relative humidity, sunshine hours and wind speed
Prior to planting, surface (0 - 20 cm) soil samples, from were used from nearby weather station.
twelve spots across the experimental field, were

The respective crop coefficient for initial, middle and after cautiously separating the grain from the straw,
late growth stages were adopted from the FAO cleaned and adjusted to 12.5% seed moisture
Penman-monteith method for wheat. Soil data like content using a hand seed moisture tester
total available water (194 mm/m), maximum rain instrument. The thousand grain weights was
infiltration rate (30mm/day), rooting depth (3m), determined by counting the number of seeds
initial soil moisture depletion (45%) and initial randomly taken from each plot and recorded on 12.5
available soil moisture (106 mm/m) were used to % moisture basis. Harvest index (HI) was calculated as
calculate crop irrigation schedule. the ratio of grain to above ground biomass yield.
Seventy percent of application efficiency was Water productivity

considered as surface method of water application was The productivity of total applied water (WP) is defined
used. Irrigation scheduling of the crop was computed as crop yield per unit volume of total water supplied
using FAO CROPWAT program (Allen et al., 1998) by (Molden, 1997).
considering soil type with fixed interval and variable
depth (refill to field capacity). Ten days fixed interval Crop yield kg-1
WP =
and refill to field capacity irrigation scheduling criteria Water supplied (P+I mm)
was adopted. Since there was no rainfall received
during the later growth stage net irrigation Where,
requirement was taken to be equal to Etc. The depth I = irrigation water (mm),
applied to other treatments was taken simply as P = the precipitation (mm) during the
percentage of the optimal irrigation. growing season.
Based on CROPWAT computer program the total Economic analysis was performed following the
amount of supplemental irrigation water for 0%, 50%, CIMMYT partial budget methodology (CIMMYT, 1988).
75% and 100% SI treatments were 0, 93, 140, 187 Economic analysis was done using the prevailing
mm, respectively. The computed depth of irrigation average market prices for inputs at planting and for
water was applied through a partial flume (7.0 cm outputs at the time of crop harvest. The average local
height) as per the treatment. First irrigation was market price of wheat was 11.29 Birr kg-1, straw 0.68
applied on 21 September based on the irrigation Birr kg-1, the cost of urea was 12.48 Birr kg-1, and
regimes. The second and third irrigation was applied application cost was 2 Birr kg-1. Labor cost for
on 29 September and 09 October 2013. The equivalent irrigation was 16.97, 12.75 and 8.51 man-days per
values for the aforementioned treatments in 2014 hectare for 100%, 75% and 50% SI treatments,
were 0, 52, 71, 95 mm and applied on 1 October and respectively. One man-day cost was Birr 100. Fuel cost
10 October. was Birr 14.08 liter-1. All costs and benefits were based
on the average of the two years. Percent MRR was
Data Collection and Measurements calculated as changes in NB (raised benefit) divided by
Days to maturity (DM): Physiological maturity was changes in cost (raised cost).
calculated by counting the number of days from 50 %
emergence to the stage when 90% of the plant reaches Finally all data were analyzed following statistical
physiological maturity. Data on productive tillers and procedures of SAS version 9.2. Whenever treatment
plant stand of the plots at harvest were recorded from effects were significant, the means were separated
randomly selected area of 0.25 m2 (0.5 x 0.5m) at four using the least significant difference (LSD) and LSD
sites and expressed as tiller population per meter fisher procedures at the probability level of( p < 0.05).
square. Plant height (PHT), spikes length (SPL),
Number of seeds per spike (NSPS) was determined Homogeneity of variances was also evaluated using
from 10 random sampled plants per plot during the F-test as described by Gomez and Gomez (1984).
physiological maturity. Grain yield and biomass yield Since the F-test has showed homogeneity of the
per plot was determined from the middle 6 rows of variances of the two years for most of the agronomic
each plot to avoid border effects. Total biomass was parameters, combined analysis was used.
determined by weighing the shoots along with the
seeds using Salter balance. Grain yield was recorded

RESULTS AND DISCUSSION that NP is inadequate for maximum crop production in

this soil, causing an important constraint to wheat
production. Therefore, the soils need amendment for
Soil physico-chemical properties successful crop production. The soil falls in category of
The soil in the study area is vertisol with clay texture non saline soils (Hazelton and Murphy (2007).
(Rowell, 1994) with a particle size distribution of 45%
clay, 30% silt and 25% sand. High clay content may Phenological and Morphological Attributes
indicate the better water and nutrient holding capacity Days to maturity was significantly (P<0.01) influenced
of the soil in the experimental site. The soil reaction is by the main effects of supplemental irrigation, N
almost neutral to slightly neutral according to the fertilization and their interaction, year of cultivation
rating of (Tekalign , 1991) indicating that it is suitable and nitrogen rates by year of cultivation interaction
for growing most crops (FAO, 2008).Wheat grows (Appendix Table I). Table 1 showed that the treatment
under a wide range of soil pH, with permissible ranges that received 100% SI level at application rates of 69
of 5.5-7.0 (Gooding and Davies, 1997). Based on the kg N ha-1 delayed maturity by 23% as compared to no
limit set by Hazelton and Murphy (2007), the soil had irrigation and no nitrogen was applied. But differences
high CEC. The data further revealed that the soil is low were not significant with the treatment that received
in available P (Cottenie, 1980) and low total N content 100% SI at application rates of 92 kg N ha-1and 46 kg N
and organic matter (Tekalign , 1991), which indicating ha-1.
Table 1. Mean values of days to maturity and plant height of bread wheat as influenced by the interaction effect
of nitrogen rates by supplemental Irrigation levels.
Supplemental irrigation N rate (kg ha-1) Days to maturity Plant height (cm)
Control (no irrigation) 0 83.50 k 64.71m
23 84.67jk 67.10l
46 85.67ij 67.53kl
69 86.33hi 69.57jk
92 86.17i 69.83j
50% CWR 0 87.67h 72.48i
23 91.00 ef 74.22ghi
46 91.83rf 83.05cde
69 90.67 f 81.95de
92 92.33e 81.12e
75% CWR 0 89.17g 73.17hi
23 90.67 f 75.37fgh
46 100.50c 82.92cde
69 102.00 ab 84.53bc
92 101.17 bc 83.47cd
100% CWR 0 96.17d 75.87fg
23 100.50 c 76.73f
46 102.83a 86.77b
69 103.00 a 89.47a
92 102.83 a 83.81cd
LSD (0.05) 1.40 2.29
CV (%) 2.58 1.26
Year
2013 76.21b 92.33b
2014 78.14a 94.53a
LSD (0.05) 0.72 0.43
Means of the same parameter in a column followed by the same letter are not significantly different at P = 0.05
according to LSD Fishers Protected Test. CWR= crop water requirement

Table 2. Mean values of days to maturity and productive tiller production of Bread wheat as influenced by the
interaction of nitrogen rate and growing Season.
Days to maturity Productive Tillers m-2
N rate(kg ha-1) 2013 2014 2013 2014
0 88.4e 89.0d 161 g 184ef
23 90.7d 92.8c 187 de 226b
46 94.2b 96.2a 194cd 222b
69 94.1b 96.9a 176f 223b
92 94.2b 97.0a 200c 258a
LSD (0.05) 1.01 9.00
CV(%) 1.26 5.50
according to LSD Fishers Protected Test.
Interaction effects among nitrogen rates and year Plant height has significantly (P<0.01) varied across
were significant. In both seasons, days to maturity was years. The highest plant height (89.47cm) was
prolonged with the increase in nitrogen rates. recorded during 2014 cropping season as compared
Significantly longest maturity (97) and 94 days was (83.80cm) to 2013. The highest plant height of wheat
recorded with application of 92 kg N ha -1 for 2014 and plant in 2014 might be attributed to relatively
2013 seasons respectively. The possible reason for adequate water which helps the plant to utilize the
long maturity in 2014 might be attributed to the better available nutrient to grow fast and taller, produce
moisture condition and nitrogen fertilizer that favored more vegetative and reproductive growth than in 2013
the growth of the crop, delayed senescence and with lower rainfall.
extended grain filling duration. While, the shortest
maturity in 2013 might be due to the moisture stress Yield components
that accelerated senescence. The results are in
agreement with Westage (2003) and Akram et al. Productive tiller:
(2004) who stated that water stress-induced The number of productive tillers m-2 was significantly
accelerated senescence after anthesis and shorten the (P < 0.01) influenced by supplemental irrigation, N
duration of grain filling by causing premature fertilization rates and their interaction as well as year
desiccation of the endosperm. A similar result on of cultivation and its interaction by nitrogen rates
cotton has been reported by Owen et al. (2011). (Appendix Table 1).
Plant height of bread wheat was significantly (P < Application of 92 kg N ha1 with 75% SI or 100% SI
0.01) varied in response to supplemental irrigation, N level increased tiller production by 94% and 92% as
fertilization and their interaction and cropping season compared to the control treatment (Table 3). This
(Appendix Table I). Table 1 showed that the treatment result is in line with Kibe et al. (2006) and Malhi et al.
that received 100%SI with nitrogen application rates (2006) who stated that increasing irrigation water
of 69 kg N ha1 increased plant height by about 39 % during heading to owering stages of wheat and
as compared to no irrigation and no nitrogen was nitrogen uptake increased tiller production of wheat.
applied. The increase in plant height in response to They also noted that stimulatory effects of N on
this treatment might be primarily due to the tillering through cytokinin synthesis are known to
contribution of higher N rates and supplementary result into more number of effective tillers of wheat. In
irrigation which improved vegetative growth, cell contrast, plants in control (140 m-2) recorded the
expansion and enlargement. This result is in lowest number of fertile tillers which might be
agreement with Ayoub et al. (1994); and Muhammad attributed to relatively low soil moisture content at the
et al. (2012) who have reported direct relationship of growth stages of anthesis and booting, which may have
nitrogen application to plant height through stressed the crop, thereby reducing survival of
elongation of internodes. productive tillers (Kibe et al., 2006).

Nitrogen rate and year of cultivation interact about 36% in comparison with no irrigation and no
significantly to affect number of productive tillers nitrogen treatment (Table 3).
(Table 2). The highest rate of nitrogen application (92
kg ha-1) increased tiller production by 24% during the Spike length has significantly (P<0.01) affected due to
2013 cropping season and by 40% during 2014 supplemental irrigation and this also varied across
cropping season compared to the control treatment. years. In this regard, the highest supplemental
irrigation level increased spike length by 46 % during
Spike length: Spike length of bread wheat was the 2014 cropping season and by 28 % during 2013
significantly (P>0.001) influenced by supplemental cropping season as compared to their respective
irrigation levels, N fertilization rates and their control (Table 4). The increase in spike length in 2014
interaction as well as cropping season and its might be associated to the better utilization of
interaction with supplemental irrigation (Appendix nutrients using the adequate moisture of the season
Table I). which helps the crop to better utilize the nutrient and
made the plant more efficient to express its potential
Spike length of wheat was significantly (P<0.01) in photosynthetic activity and influence to increase
affected by the interaction effect of nitrogen rates and length of spike. Attia and Barsoum (2013); and Shirazi
supplemental irrigation levels. The treatment where et al. (2014) also indicated that irrigation resulted in
nitrogen was applied at the rate of 69 kg N ha-1 and increased spike lengths of wheat compared with non-
irrigated with 100 % SI level increased spike length by irrigated treatment.
Table 3. Mean values of productive tiller, spike length, seeds per spike, 1000 kernel weight and harvest index as
influenced by nitrogen rates and Supplemental Irrigation levels interaction
Supplemental irrigation N levels kg ha-1 Productive Spike Seeds per Thousand
Levels Tiller length Spike(no) seed weight
m-2 (cm) (g)
Control (no irrigation) 0 140.00 k 7.23i 31.13 m 28.56j
23 150.00k 7.37i 33.63l 29.52j
46 177.67h 7.73ghi 36.31 ij 31.32i
69 164.33ij 7.73ghi 35.27jk 29.16j
92 177.10 hi 7.77ghi 34.01 kl 29.32j
50% CWR 0 163.33j 7.61hi 35.01 jkl 32.91h
23 206.67de 8.51ef 38.73gh 34.92g
46 211.67de 9.17d 39.81 fg 35.93fg
69 228.34c 8.81de 41.67 de 36.42ef
92 236.65bc 8.93de 41.91de 36.07fg
75%CWR 0 203.33def 7.98fgh 37.28 hi 36.03fg
23 200.00ef 8.95de 39.58g 37.67de
46 201.65 def 9.85c 41.62 de 39.05cd
69 191.64fg 9.95bc 42.91 cd 40.11bc
92 275.00a 10.15bc 43.47c 39.98bc
100%CWR 0 183.00gh 8.25fg 38.97 g 37.47e
23 227.00c 10.45b 41.21ef 39.73bc
46 241.76 b 11.25 a 43.91 c 41.82a
69 213.33d 11.58 a 45.96 b 41082a
92 270.00a 10.42b 47.59a 42.02a
LSD(0.05) 13 0.54 1.53 1.38
CV% 5.50 4.52 3.01 2.84
Year
2013 183.46b 8.09b 38.09b 34.86b
2014 222.76 a 9.87a 40.89 a
40.89a
LSD(0.05) 4.06 0.15 0.43 0.37

Table.4. Mean values of spike length, number of seeds per spike and biological yield of bread wheat as influenced
by the interaction of supplemental irrigation by year of cultivation, Enderta
Spike length (cm)
Supplemental irrigation 2013 2014
0 (rain-fed) 7.07f 8.06de
50% CWR 7.93 e 9.27c
75% CWR 8.33d 10.43b
100% CWR 9.04 c 11.74a
LSD (0.05) 0.29
CV(% ) 4.40
Number of kernels per spike
0 (rain-fed) 32.47e 36.80d
50% CWR 38.09 cd 42.74b
75% CWR 39.21c 44.23ab
100% CWR 44.04 b 46.47a
LSD (0.05) 2.27
CV(% ) 3.10
Thousand kernel weight (g)
0(rain-fed) 28.07g 31.07f
50% CWR 33.99e 36.50d
75% CWR 37.56 c 39.57b
100% CWR 39.83 b 40.99a
LSD (0.05) 0.87
CV(%) 6.00
Aboveground biomass yield ( kg ha-1)
0(rain-fed) 3045.7f 3681.1e
50%CWR 5187.6d 6493.5c
75%CWR 7534.3 b 8654.7a
100%CWR 7940.0 b 9002.7a
LSD (0.05) 494.78
CV(% ) 9.34
Means of the same parameter in a column followed by he same letter are not significantly different at P = 0.05
according to LSD Fishers Protected Test. CWR= crop water requirement
Number of Seeds per Spike (NSPS): of both nitrogen and water in the soil which may have
Number of seeds per spike of bread wheat was increased floret endurance whilst the reduction in the
significantly (P<0.01) influenced by supplemental control plots might be attributed to deficiency of
irrigation levels, N fertilization rates and their assimilates due to water stress during grain filling
interaction as well as cropping season and its (Ferrante et al., 2010), reduced number of grains per
interaction with supplemental irrigation (Appendix unit area due to flower abortion (Acevedo et al., 2002)
Table I). and reduces pollination Shraf (1999).
The number of seeds per spike were significantly Kernel per spike were significantly (P<0.01) affected
(P<0.01) affected by the interaction effect of nitrogen by supplemental irrigation, which also varied across
rates and supplemental irrigation levels. The years. In this regard, the highest number of kernels per
treatment where nitrogen was applied at the rate of 92 spike increased kernels per spike by 46% during the
kg N ha-1 and irrigated with 100 % SI level increased 2014 cropping season and by 35% during 2013
seeds per spike by 51 % as compared to no irrigation cropping season as compared to their respective
and no nitrogen was applied (Table 3). The increase in control (Table 4). The result are in agreement with the
number of grains per spike might be due to availability findings of Madani et al. (2010) ; and Passioura and

Angus (2010) who reported that stress after anthesis and no nitrogen was applied in 2014 and 2013
reduced pollination, size of the individual grains and cropping season, respectively. The highest thousand
the grain number through abortion of the developing kernel weight of wheat in 2014 might be attributed to
grains which resulted in the reduction in grain yields. relatively adequate water which helps the plant to give
extra time to translocate the carbohydrate to grains
Thousand Kernel Weight (TKW): which improves grain size than in 2013 with lower
The TKW of bread wheat was significantly (P < 0.01) rainfall (Zhang and Oweis, 1998).
influenced by supplemental irrigation levels, N
fertilization rates and their interaction as well as Aboveground biomass and Grain Yields
cropping season and its interaction with supplemental Total aboveground biomass responded significantly (P
irrigation (Appendix Table1). < 0.01) to supplemental irrigation, N fertilization rates
and their interaction as well as cropping season and its
The treatment where nitrogen was applied at the rate interaction with supplemental irrigation (Appendix
of 92 kg N ha-1 and irrigated with 100% SI level, Table 1). Table 5 showed that increasing the rate of
increased TKW by 47% as compared to no irrigation nitrogen application from 0 to 92 kg N ha -1 and
and nitrogen application but this treatment was in supplemental irrigation from zero to 100% had
statistical parity with the treatment that received 69 increased aboveground biomass yield of the wheat
kg N ha-1 of the same SI level (Table 1). The increase in plants consistently. The increases in the aboveground
thousand kernels weight could be attributed to higher biomass yield obtained in response to application of
grain weight of well-watered plants which could be 100%, 75% and 50%, SI irrigation combined and 92 kg
associated with longer grain filling duration and faster N ha-1 over the control treatment were 340%, 58%
grain filling rate as reported by Li et al. (2000). These and 7%, respectively. However, the aboveground
results are in agreement with the findings of Zhang et biomass yield obtained in response to application of
al. (1998) who also reported that adequate water at 100% irrigation and 92 kg N ha-1 were in statistical
anthesis not only allowed the plant to increase parity with the aboveground biomass yield obtained in
photosynthesis rate but also gave extra time to response to the application of 100% irrigation and 69
translocate the carbohydrate to grains which kg N ha-1. Similar findings were reported by Tavakoli
improved grain size and yield. et al. (2010) who found that irrigation and nitrogen
significantly enhanced biomass yield of wheat, in
The reduction in thousand kernel weight in response which case no significant differences were observed in
to decreased irrigation may be attributed to water biomass production between 66% and 100%
shortage effects in seed filling period which cause to irrigation levels and between nitrogen fertilizer rates
the reduction of absorption and translocation of of 60 and 90 kg N ha-1.
water and nutrients in plant and reduction of
nutrients translocation rapidity to seeds. This The highest aboveground biomass yield obtained at
suggestion is consistent with the findings of Begdelo et 100% irrigation combined with 69 and 92 kg N ha -1
al. (2011) who stated that thousand kernels weight might be attributed to the synergistic effect of
was reduced under moisture stress conditions. improved N nutrition and enhanced moisture content
Thousand kernel weight has significantly (P<0.01) of the soil on growth and photosynthetic efficiency of
influenced by the interaction of supplemental the plants, enhanced tillering and delayed senescence
irrigation levels and year of cultivation. The highest of leaves of the crop which may have resulted in higher
TKW (41g) was recorded during 2014 cropping season production of dry matter. Moreover, there were no
with the application of 100% SI level as compared further statistical yield increases beyond 46 kg ha-1
(39.8g) to 2013. In this regard, the year explained 3 % under non-irrigated plots and 50% SI plots. This could
total variability on thousand kernel weight of wheat be possibly due to supra-optimal rates of nitrogen
(Table 4).In both seasons, thousand kernels weight supply to the plant that was above optimum under
progressively increased with increasing the level of moisture stressed condition.
supplemental irrigation. The lowest TKW was
recorded in 2013 than that of 2014 cropping season. Aboveground biomass yield varied significantly in
The treatment that received 100% SI level increased response to supplemental irrigation as well as across
TKW by 31 % and 42% as compared to no irrigation the years (Table 5). In both years, aboveground
biomass yields increased with the increase in SI level.

A higher aboveground biomass yield was recorded Grain Yield (kg ha-1):
during the 2014 cropping year than during the 2013 The grain yield of wheat responded significantly (P <
possibly due to the favorable moisture conditions 0.01) to the supplemental irrigation levels, N
during growing period that may have improved the fertilization rates and their interaction and year of
efficiency of nutrient utilization, and thus the yield cultivation (Appendix Table1). Grain yield increased
attributing characters and yield of the crop. significantly (P < 0.01) with the increased level of
irrigation and nitrogen being the highest in plots that
The magnitudes of increase in response to 100% received 100% SI with 92 kg N ha1 and the lowest in
irrigation over the rain-fed, 50% and 75% SI treatme- plots that received no irrigation and nitrogen (Table
nts were 160%, 53%, and 5%, respectively, in 2013. 5). Keeping the N fertilization at 69 kg ha -1 and
The equivalent values for the 2014 cropping year were applying 100%, 75% and 50% irrigation levels
144%, 39% and 4%, respectively. This may be attribu- increased grain yield by 408%, 75% and 9%,
ted to the differences in the amount of rainfall received respectively over none irrigated control. However,
during the growing season. Thus, the rainfall of only there was no significant difference between the
317 mm during the growing season of 2013, which treatment that received 69 kgN ha-1 and 92 kg N ha-1 at
was as much as 537 during the growing season of 100% SI which increased the grain yield by about
2014, may have made the crop respond to irrigation 385% over the control treatment.
more vigorously in the former than the latter growing
seasons.
Table 5. Mean values of biological yield , grain yield and water productivity of wheat as influenced by nitrogen
rates by Supplemental Irrigation levels interaction.
Supplemental N levels Biological yield Grain yield W P kg ha-1 mm-1
Irrigation kg ha -1 (kg ha )
-1 kg ha )
-1
Control 0 2505.11j 885.91l 2.15j

23 3046.10ij 1087.81kl 2.67hij
46 3774.10ghi 1367.10ijk 3.39efg
69 3987.85fgh 1461.61hij 3.58ef
92 3504.75ghi 1253.62ijk 3.06fgh
50% CWR 0 3343.42hi 1189.61jkl 2.39ij
23 5110.01e 1886.42g 3.80e
46 6593.24d 2478.56f 5.03cd
69 7171.01d 2715.01f 5.50c
92 6985.33d 2595.91f 5.24cd
75% CWR 0 4221.71fg 1543.65hi 2.90ghi
23 6937.81d 2596.71f 4.97cd
46 8274.22c 3228.01e 6.17b
69 9831.05b 3987.52cd 7.42a
92 10209.42b 4119.31bc 7.60a
100%CWR 0 4751.11ef 1713.11gh 3.00fghi
23 7009.32d 2643.56f 4.68d
46 9010.91c 3630.01d 6.42b
69 10563.56ab 4297.03ab 7.60a
92 11023.65a 4505.85a 7.99a
LSD(0.05) 765.80 322.70 0.63
CV% 9.34 10.39 10.64
Year
2013 5976.90b 2306.00b 5.26a
2014 6808.02a 2602.04a 4.32b
LSD(0.05) 217.36 92.87 0.18
Means of the same parameter in a column followed by the same letter are not significantly different at P = 0.05 according to
LSD Fishers Protected Test. TKW=thousand kernel weight, CWR= crop water requirement, PAW = Productivity of available
water .

This suggests that the extra N levels greater than 69 kg insufficient, fertilizer nitrogen was inadequately taken
N ha-1 had little influence on increasing yield. The by the plant and some of it may have been lost by
possible reason of increased yield at 100% denitrification in an experiment involving a
supplemental irrigation and nitrogen rates at 69 kg N wheat/sunflower cropping sequence.
ha-1 might be largely attributed to the effect of
nitrogen in sustaining leaf photosynthesis activity, The grain yield produced in response to the
delaying leaf senescence during grain filling period application of 69 kg N ha-1 combined with 100%
and extended duration of grain filling when moisture irrigation, which was agronomically optimum in this
supply was not limited. Likewise, supplementary experiment, was five-fold higher than the grain yield
irrigation upon the cessation of rains may have produced in response to the rainfed treatment.
allowed remobilization of photoassimilates from Similarly, this yield was 4-fold, 3-fold, 3-fold, and 3.4-
source to sink as was evidenced by Abderrazzak et al. fold higher than the grain yield produced without
(2013). The results are in conformity with the findings irrigation but with combined applications of 23, 46, 69,
of other researchers (Abbas, 2010; Tsegay et al., 2015; and 93 kg N ha-1 in order mention here. In this
Yihun et al., 2013) who reported supplemental connection, the blanket recommended rate of N
irrigation at heading; anthesis and early grain filling fertilizer in the study area is about 41 kg N ha -1and 46
period resulted in maximum wheat and tef yields. This kg P2O5 (100 kg DAP and 50 kg Urea per hectare)
finding is also supported by Tadayon et al. (2012) who (Elias, 2002). However, farmers may be applying much
showed that supplemental irrigation at jointing stage less than even the blanket recommended rate of the
produced greater grain yield and higher water fertilizers possibly because of escalating fertilizers
productivity with application of 100 kg N ha-1. prices (Bekunda et al., 2010), uncertainty of response
to fertilizers because of frequent moisture stress
The lowest grain yield obtained in the control during the growing season (Morris et al., 2007). As a
treatment (885 kg ha-1) might be attributed to reduced result, the average grain yield of bread wheat obtained
production of photosythate because of deficiencies of by smallholder farmers under rain-fed condition in the
both nitrogen and moisture, which may have resulted study area is about 1.5 t ha-1 (TBOARD, 2013), which is
in stomatal closure and early senescence that could almost equal to the yield obtained under rain-fed
have ultimately affected grain development processes. condition (no irrigation), combined with the
Similar results were reported by Akram et al. (2004) application of 69 kg N ha-1 in this study. Thus, the
who stated that water stress-induced accelerated average grain yield obtained by smallholder farmers in
senescence after anthesis shortening the duration of the study area is only about 1/3rd of the grain yield
grain filling and causing premature desiccation of the obtained in this experiment in response to the
endosperm and reducing embryo volume, grain yield, application of 100% supplementary irrigation
and kernel weight. combined with 69 kg N ha-1. This result reveals that
farmers in the study area are losing about 66% wheat
The magnitude of decrease in grain yield in response grain yield to moisture stress under the rain-fed
to increasing nitrogen from 69 kg N ha-1 to 92 kg N ha-1 production system although the blanket recommended
under non-irrigated treatment was 14%. A similar rate of fertilizer they apply on average to the crop
result was reported by Tavakoli et al. (2010) who seems adequate.
described that the lowest response to N was under
rainfed conditions and the highest level (120 kg.N.ha-1) The results are in line with the findings of Alemtsehay
tended to decrease yields more. While, response to N et al. (2015) who reported that uptake of N and P by
were not limited by available water at the 2/3 of full tef [Eragrostis tef (Zucc.) Trotter] was enhanced in
irrigation level, as they were similar to those at full response to increasing the rates of the fertilizers in the
irrigation. presence of supplemental irrigation. Araya and
Stroosnijder (2011) also reported that more than 80%
The results of this study are also corroborated by the of the yield reduction and more than 50% of crop
findings of Thomson and Whitney (1998) who showed failures in tef and barley can be avoided if SI is
that the most important factors controlling the effect employed during the critical growth stages of the
of nitrogen on crops is water supply. Corbeels et al. crops.
(1998) showed that when water supply was

Similarly, Oweis and Hachum (2003) found that wheat of this research indicated that water and nitrogen use
yields increased from 2 tons per hectare to more than efficiency was greatly increased by supplemental
5 tons per hectare in response to the use and timely irrigation. Similar results were reported by (Zhang et
application of only 100 to 200 mm of irrigation water. al. 2004; Ali et al. 2007; Oweis et al. 2010).
SI increased the yield of tef from less than 1 t ha -1 to 2
tons ha-1 (Alemtsehay et al., 2015). Similarly, Ashraf et In both seasons, water productivity decreased with the
al. (2001) reported that when water was applied at increase in SI level up to 100 % SI. Significantly higher
mid grain fill pearl millet produced 10 % more grain water productivity was observed during 2013
yields than with no irrigation. cropping season (6.54 kg ha-1 mm-1) when the crop
was supplemented with 75% SI level as compared to
Water Productivity (WP) (5.58 kg ha-1 mm-1) during 2014 crop season (Table 6).
The productivity of total applied water (PAW) In this regard, 75% SI level increased WP by 161%
responded significantly (P 0.01) to supplemental during 2013 cropping season as compared to 129 % in
irrigation levels , N fertilization rates and their 2014 cropping season. However, increasing the rate of
interaction, year of cultivation and the interaction supplemental irrigation from 75% to 100% levels
effect of supplemental irrigation levels by year and decreased the water productivity by 4 % during the
nitrogen rates by year of cultivation(Appendix Table 2013 cropping season and by about 7 % during the
I). Table 5 showed that increasing the rate of nitrogen 2014 cropping season, respectively.
application from 0 to 92 kg N ha -1 with supplemental
irrigation from zero to 100 % SI significantly (P < The results showed a more significant improvement in
0.001) increased water productivity (7.99 kg ha -1 mm- SI water productivity at SI level application rates
1) of the crop. Thus, increasing the rate of nitrogen than at full SI level. The increase in water productivity
from 0 N ha-1 to 92 kg N ha-1 and no irrigation to 100 might be attributed to the effectiveness of a small
% SI increased WP of the crop by 271% over control. amount of water in alleviating severe moisture stress
There was however, no statistical difference with plots during the crop development and grain filling. While,
that received 100 % SI level with 69 kg N ha -1 (7.60 kg the decrease in water productivity at the high SI levels
ha-1 mm-1) , 75% SI level at application rates of 92 kg N might be due to increase grain yield is less than
ha-1 (7.84 kg ha-1 mm-1) and 75% SI level at application proportional increase in ET. Similar results were
rates of 69 kg N ha-1 (7.41 kg ha-1 mm-1). The findings reported by (Maliheh and Aliasghar, 2011).
Table 6. Interaction effect of supplemental irrigation and year and nitrogen rate and year on water productivity in
Enderta , Tigray
WP (kg ha-1 mm-1)
Supplemental irrigation 2013 2014
0 (rain-fed) 2.50f 2.43g
50% CWR 4.69 d 4.09e
75% CWR 6.54 a 5.58b
100% CWR 6.30a 5.18c
LSD (0.05) 0.40
Nitrogen (kg N ha-1)
0 2.70f 2.51f
23 4.48d 3.58e
46 5.85 b 4.65d
69 6.63 a 5.45bc
92 6.61a 5.41bc
LSD (0.05) 0.41
CV(% ) 10.64
according to LSD Fishers Protected, PAW = Productivity of available water, CWR= crop water requirement

Table 7. Mean marginal rate of return on the effect of nitrogen and supplemental irrigation levels on wheat in
Enderta district, Tigray
Supplemental Applied N TCV Net benefit Raised cost Raised benefit MRR%
irrigation (CWR %) kg ha-1 (ETB )ha-1 (ETB ha-1) (ETB ha-1) (ETB ha-1)
0 0 0.00 10495.0 0.0 0.00 0.00
0 23 724 12144.5 1649.5 724.0 227.83
50 0 1291 12789.3 644.8 567.0 113.72
75 0 1935 16264.3 3475.0 644.0 539.60
50 23 2015 20195.4 3931.1 80.0 4913.88
75 23 2659 27855.3 7659.9 644.0 1189.43
75 46 3333 30732.3 2877.0 674.0 426.85
100 46 3975 38260.9 7528.6 642.0 1172.68
75 69 4007 43658.9 5398.0 32.0 16868.70
100 69 4649 45284.7 1625.8 642.0 253.24
100 92 5323 47004.7 1720.0 674.0 255.19
CWR= crop water requirement, TCV= total cost that vary, MRR= marginal rate of return, ETB= Ethiopian Birr
In general higher water productivity was observed in Partial Budget Analysis

2013 compared to 2014 cropping season. The results Treatments having marginal rates of return (MRR)
are in close collaboration with Magombeyi et al. below 100% were considered low and unacceptable to
(2009) who found large yield responses to farmers, thus, eliminated (CIMMYT, 1988). This was
supplementary irrigation of maize, ranging from 67 to because such a return would not offset the cost of
314 %, with the highest responses in the driest years. capital and other related costs while still giving an
They also signify that the significant response of attractive profit margin to serve as an incentive. The
supplementary irrigation even in the wettest year maximum net benefit of 47004 ETB ha-1 with optimum
reflects the erratic rainfall pattern typical of the semi marginal rate of return was recorded at the rate of 92
arid tropics, and the potential for supplementary kg N ha-1 supplemented with 100% irrigation, followed
irrigation to bridge dry spells at critical stages .This by nitrogen application rates of 69 kg N ha -1
study is also in line with Rahman et al. (2000) who supplemented with 100% irrigation (Table 7).
reported that WP is the highest in low rainfall season
and gradually decreased with increasing irrigation
levels in good season. CONCLUSION
In both seasons, water productivity decreased when The results of this study have demonstrated that grain
nitrogen level was increased from 69 to 92 kg N ha -1. yield, yield components and water productivity of
Significantly higher water productivity was obtained bread wheat were strongly influenced by
in 2013 (6.63 kg ha-1 mm-1) at application rates of 69 N supplemental irrigation and nitrogen application.
kg ha-1 as compared to (5.45 kg ha-1 mm-1) in 2014. In Maximum bread wheat grain yield of 4504 kg ha-1 the
this regard, the increasing rate of application from 69 highest water productivity 7.99 kg mm-1 ha-1 was
N kg ha-1 to 92 N kg ha-1 decreased water productivity recorded in response to the application of 100%SI
by 4 % during the 2013 cropping season and by about combined with 92 kg N ha-1. However, the lowest grain
7 % during the 2014 cropping season, respectively. yield of 885 kg ha-1 and the lowest water productivity
Decreased productivity at higher rates was also of 2.15 kg mm-1 ha-1 were recorded in response to the
reported by Oweis and Hachum, 2003. Moreover, the rain-fed production with no application of any
application of 69 kg N ha-1 increased water nitrogen fertilizer. Based on economic analysis, the
productivity by 145 % during the 2013 cropping acceptable marginal rate of return of 255% and the
season and by 117% during 2014 cropping season as highest net benefit (47004 ETB ha-1) was obtained
compared to control. from the application of 92 kg N ha-1 with 100%

supplemental irrigation. The results of the study Regimes. International Journal of Science and Advanced
indicated that in order to improve the grain yield and Technology, 1(9):120-125.
CIMMYT (International Maize and Wheat improvement
yield components of wheat grown in the clay soils of Center).1988. From Agronomic Data to Farmers
Enderta farmers should adopt supplementary Recommendations: Economics Training Manual.
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Clarke D, Smith El-Askari K (1998) Users Guide:
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Corbeels M, Hofman G and Cleemput O (1998) Residual
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cropping sequence on a Vertisol under semi -arid
Mediterranean conditions.European Journal of
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Appendix Table I. Analysis of variance for phonology, growth characteristics, yield and yield components and
water productivity of wheat as affected by , supplemental irrigation and nitrogen rates at Enderta, 2013 and 2014
cropping seasons.
MSE of agronomic variables

Source of variation DF DM PHT PTM SPL NSPS TKW BYLD
Replication 2 15.108 32.85 312.93 2.98 59.8 37.63 224802
SI 3 1431.04** 1268.14** 24419.41** 42.87** 478.6** 678.85** 160575812**
Error1 6 2.7194 3.9176 89.38 0.875 6.115 6.196 1491320
NL 4 202.57** 467.19** 9903.13** 13.02** 158.05** 45.265 77079680**
SI.NL 12 36.86** 27.34** 3288.58** 1.53** 7.94** 4.0391 5632169**
Error 2 32 2.3792 8.90525 249.89 0.309 2.9044 2.33877 293751
Year 1 145.2** 111.16** 46334.7** 95.40** 233.80** 140.83** 20722115**
SI.year 3 1.6222 0.05764 15.14 4.39** 1.30** 4.58** 1445703*
NL.year 4 2.1** 0.0125 1212.2** 0.033 0.300 0.113 201833
SI.NL.year 12 0.0389 0.0125 28.76 0.033 0.300 1.432 156353
Error 3 40 0.60 0.0271 24.70 0.050 0.229 1.042 406979
Total 119
DM= days to maturity, PHT= plant height ,PTM= productive tiller m2 , SPL=spike length ,NSPS= number of seeds per spike , BYLD=biological yield, GYLD= grain yield ,HI=
harvest index, PAW= productivity of available water.
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RESEARCH ARTICLE
Community structure and monthly dynamics of zooplankton in

high altitude rice fish system in Eastern Himalayan region of India
Saikia Rajashree1,2*, Das Tapati1, Gogoi Budhin2, Kachari Akash2, Safi Vivekanand2 and Das Debangshu N2
1Department of Ecology and Environmental Science, Assam University, Silchar, India - 788011
2Department of Zoology, Rajiv Gandhi University, Rono Hills, Doimukh, Arunachal Pradesh, India 791112
*Corresponding author: Rajashree Saikia, Department of Ecology and Environmental Science, Assam University
Silchar, India - 788011, Contact No. +91-9854474964, Email ID: rajashreesaikia1@gmail.com

Received: 19.07.2017 The community structure and seasonal dynamics of zooplankton were
Accepted: 20.08.2017 examined in high altitude rice fish system of Eastern Himalaya. The study
Published : 19.09.2017 was conducted at fortnight interval from field preparation to harvesting of
rice during two seasons of 2013 and 2014. All the standard procedures
Editor:
were followed to obtain relevant quantitative and qualitative data on
Dr. Arvind Chavhan
zooplankton showing, a total of five major communities of zooplankton viz.,
Cite this article as: Cladocera, Copepoda, Rotifera, Protozoa, and Ostracoda were encountered.
Saikia Rajashree, Das Tapati, Gogoi These comprised of 33 taxa of Cladocera, 11 taxa of Copepoda, 17 taxa of
Budhin, Kachari Akash, Safi Rotifera, 5 taxa of Protozoa and only one taxon of Ostracoda. Cladocera was
Vivekanand and Das Debangshu N the most dominant (29%) while Rotifera (12%) was least abundant.
(2017) Community structure and Density of zooplankton communities showed significant (p<0.05%)
monthly dynamics of zooplankton in monthly variation during the rice growing seasons. The computed diversity
high altitude rice fish system in
indices confirmed the presence of diversified zooplankton in the rice fish
Eastern Himalayan region of India;
system. Month wise cluster of similarity index revealed very close species
362-378. composition at initial and mid phases compared to end phase of the rice
growing season. The zooplankton therefore becomes natural food sources
of fishes concurrently in the flooded rice fields to enhance unit land
Copyright: 2017| Author (s), This productivity.
terms of the Creative Commons Keywords: Abundance; Diversity; Dynamics; Richness; Zooplankton.
provided the original work is INTRODUCTION
commercial and no modifications or Rice fish farming system is a very famous and mostly studied seasonal
adaptations are made. aquatic ecosystem for productivity of rice and fish together (Frei et al.
2007) and is recognized as a globally unique agro ecosystem in tropical and
sub tropical Asia (Lu and Li, 2006). Though wet rice fields covers the entire
spectrum of diverse seasonal freshwater fauna (Fernando, 1977; 1993);
however, assessment of zooplankton diversity, species richness, seasonal
dynamics, abundance, distribution and ecology was reported till date from
Malaysia, India, Sri Lanka, Japan, Philippines, Thailand, Indonesia and
Italian rice fields only (Bambaradeniya and Amarasinghe, 2003; Meijen

Community structure and monthly dynamics of zooplankton in high altitude rice fish system
1940; Neale, 1977; Heckman, 1979; Lim et al. 1984; organisms flourish on the energy received from
Ali, 1990; Simpson et al. 1994; Taniguchi et al. 1997; primary producers and become readily available to the
Leoni et al. 1999; Rossi et al. 2003; Chittapun et al. different categories of consumers in the rice fields
2009; Bahaar and Bhat, 2013). including stocked young carps. Therefore, fish growth
and survival is directly influenced by the abundances
In India, the Apatani hill tribes of Eastern Himalaya are and diversity of zooplankton which are involved with
habituated in raising fish concurrently as companion the process of decomposition and circulation of both
crop of rice as their main food source in high rainfall allochthonous and autochthonous organic matters
mountainous zone since last 1960 years (Kacha, (Shil et al. 2013; Guangjun, 2013). The micro
2016). This ecofriendly and economically beneficial crustaceans, rotifers and protozoans in the field water
farmers practice has made the system unique in the thrive on the dissolved organic matters and thus make
context of aquatic resource utilization (Singh et al. the system potentially favorable for the rice field
2011). The farmers sometimes use household and fishery development (Lemly and Dimmick, 1982;
agricultural wastes, excreta of domestic animals like Saikia and Das, 2010).
pig, cow and goat in the fields as energy subsidy
making such farming more sustainable and organic in From the above perspective, goal of the study is to
nature (Saikia and Das, 2004). They also cultivate the explore the zooplankton community structure and
fields dykes with various nitrogen fixing vegetables their seasonal population variability so that judicious
and millets for enhanced productivity. Regarding fish strategies of natural resource exploitation may be
species, three strains of Common carp viz., Cyprinus developed for better rice fish farming in future.
carpio specularis, C. carpio communis, C. carpio nudus
are used to stock just after water accumulation in to
the rice fields (Das et al. 2007). These stocked fishes MATERIALS AND METHODS
practically depend on the natural food sources of the
rice fields and farmers hardly use any supplementary Study site
fish feed (Ali, 1988; Saikia and Das, 2009). The micro
crustaceans, Rotifers and Protozoans in the field water
thrive on the dissolved organic matters making the
system potentially congenial for rice field fishery
(Lemly and Dimmick, 1982; Saikia and Das, 2010).
Within a crop cycle, therefore the Apatani farmers
could harvest 500.0 kg/ha of fishes and 3.0 tons/ha of
rice in an average from their rice fish system (Saikia et
al. 2015; Li, 1988).
Zooplankton being the naturally available fish food

organisms in high altitude rice fields seems to have
influencing role towards the development of rice field
fishery. The frequently changing nature of rice fields
according to the different growth stages of rice
provide congenial habitat for rich number of
diversified organisms (Heckman, 1979; Simpson and
Roger, 1995; Simpson et al. 1993). These organisms
enter into the field water via irrigation water and air to
colonize on different microhabitat of rice fields
(Fernando, 1995; 1996). The vertebrate fauna like
amphibians, reptiles, birds, mammals and the variety
of invertebrate fauna including zooplankton in rice
fields can inhabit different drastic agronomic changes
of rice field within a very short period of time Figure 1. Location Map of the study sites (Redrawn
(Bambaradeniya et al. 2004). The zooplanktonic after Saikia et al. 2015)

Saikia et al., 2017
Three high altitude (1800 m above the mean sea level) and Buzas Gibsons evenness index (Buzas Gibson,
rice fish culture plots were selected randomly from 1969).
five villages (Figure 1) namely Pine grobe, Tajang,
Nenchalya, Dutta, Mudang tage (altitude:, latitude: All the equations related to the above mentioned
2650'- 9821' N and 9240' - 9421' E longitude) in diversity indices are mentioned below:
Eastern Himalaya.
a) Shannon-Wiener diversity index
Field sampling and Analysis
Field sampling started fortnightly from April to
September during the two consecutive rice growing
seasons of 2013 and 2014 and data were presented
[Where, ni= number of individuals of taxon i, n= Total
separately for both the seasons. Daily rainfall (mm)
number of individuals]
was recorded from the nearest meteorological station
of the plateau. Water parameters like water
b) Simpsons dominance index
temperature (WT), water depth (WD), electrical
conductivity (EC), total dissolved solids (TDS) and pH
were recorded using thermometer (ZPHI-9100 Zico,
India Ltd), centimeter scale and digital water analysis
kit (SYSTORNICS 371) respectively. On the other hand, [Where, D= Simpsons dominance index; ni= number of
dissolved oxygen (DO), free carbon di oxide (FCO2), individual of taxon i, N= total number of individuals in
nitrate-nitrogen (NO3-N), phosphate-phosphorus (PO4- each species]
P), total alkalinity (TA) and chloride (Cl -1) were also
estimated (APHA, 2012). Plankton were collected by c) Buzas-Gibson evenness index
filtering 25 L of field water through standard mesh
(size 35 m) and immediately preserved in 4%
formalin. Furthermore, the preserved samples were
[Where, Buzas-Gibson evenness index; eH' =
analyzed using light microscope (Nikon, ECLIPSE
E200, Olympus CX4 and Leica DM 5000). The
quantitative analyses of zooplankton were done Shannon-Wiener index, calculated using natural
following drop count method (Lackey, 1938). logarithms, S = Number of species]
Identification of individual zooplankton was d) Margalef richness index
performed following standard keys and monographs
of Smirnov (1971, 1996); Needham and Needham,
(1972); Tonapi, (1980); Michael and Sharma (1988);
Battish, (1992); Edmonson, (1992); Shiel (1995); [Where, Margalef richness index;
Subhash Babu and Nayar (2004), Kotov et al. (2012;
2013); Sharma and Sharma (2013) and related other S= Number of species, N= Total number of individuals
related published taxonomical works on various in the sample].
species mostly up to genus level because of intricacy in
species confirmation. Statistical analysis
All the statistical analyses were carried out by using
Relative abundance [% RA = (n/N) 100] was the softwares, PAST version 3.07, SPSS version 21 and
computed to generate the percentage composition of Microsoft excel. The month-wise population
zooplankton community where, n = total number of fluctuations of various zooplankton species were
the individual per species under consideration; N = examined using Kruskal-Wallis test. Further, the
total number of all individuals per station. Attempt has similarities in species composition of zooplankton
also been made to find out the diversity of communities within months were assessed through
zooplankton in the rice field using various indices like the Bray-Curtis cluster analysis. The multiple
Shannon-Wiener diversity index (Shannon and regression analysis of physico-chemical parameters of
Wiener, 1949); Simpsons dominance index (Simpson rice field water for the zooplankton belonging to
(1949); Margelefs richness index (Margelef, 1968) different communities was done.

RESULTS AND DISCUSSIONS well as diverse zooplankton and the findings also
matched with the observation of Ali (1990) from
flooded rice field in Malaysia. The presence of aquatic
Community structure: Abundance and species weeds, decomposition of detritus and shallow depth of
composition rice field water (Sharma et al. 2012) might be reason
The findings clearly revealed that zooplankton for dominance of Cladocera in the rice field of the
community structure showed variations in terms of study area. The incident sunlight and abundant algal
species composition, density, diversity, dominance, communities supported the rapid growth of filter
richness and evenness with respect to different feeding microcrustaceans like Daphnia sp. and
months. The variations in physico-chemical Bosmina sp. in the initial rice growing cycle. Besides,
parameters of rice field water seem to be the major several biotic and abiotic interactions might selectively
contributor for seasonal abundance of zooplankton in act on some detritivorous zooplankton like Alona sp.
shallow wet rice field situated even in higher altitude. and Chydorus sp. where Chydoridae became most
The mean value of water physico-chemical parameters dominant among Cladocerans was in conformity with
like pH (6.49), WT (27.30C ), WD (16.5 cm), EC the findings of Shah et al. (2008). The periphytic
(523.62 mgl-1), TDS (302.45 mgl-1), DO (6.32 mgl-1), associations perhaps influenced the high abundances
FCO2 (12.05 mgl-1 ), NO3-N (1.34 mgl-1), PO4-P (0.14 of the members of Macrothricidae family in the rice
mgl-1), TA (29.47 mgl-1), Cl-1 (36.15 mgl-1) were fields (Sharma and Sharma, 2014). In addition to
analyzed and average rainfall (615.95 mm) was also available nutrients, the alkaline water of rice field
recorded during the whole study period (Table 1). perhaps became one of the reasons for the dominance
of Cyclops sp. (Kumar et al. 2004; Ahmad et al. 2011;
A total of five major communities of zooplankton viz., Bhat et al. 2014).
Cladocera, Copepoda, Rotifera, Protozoa and
Ostracoda were encountered in the inundated rice The Bray Curtis cluster analyses depicted that the
fields of Apatani Plateau during the rice growing monthly species compositions of zooplankton in the
season. These comprised of 33 taxa of Cladocera, 11 water logged rice field showed more similarity
taxa of Copepoda, 17 taxa of Rotifera, 5 taxa of between April and May, as represented by one cluster,
Protozoa and only one taxon of Ostracoda (Table 2). another cluster was represented by June, July and
Cladocera were represented by family Bosminidae, August, the third cluster was represented separately in
Chydoridae, Macrothidae, Daphniidae and Moinidae. September (Figure 3).
Copepoda were represented by family Cyclopidae,
Diaptomidae, Centropugidae and Canthocamptidae. Table 1. Mean value of water physico-chemical
Ostracoda was represented by a single family parameters among months in high altitude rice
Cyprididae. Among the Rotifers, the families were wetlands of Apatani Plateau during the rice growing
Lecanidae, Brachionidae, Testudinellidae, season of 2013-14 (n=45).
Dicranophridae, Notommatidae and Asplanchnidae Parameters Mean SD
whereas family Arcellidae, Centropyxidae, Difflugiidae
and Ophryoglenidae were found under Protozoa. The
pH 6.490.265
Cladoceran community was mostly dominated by
WT (C ) 27.302.5
Bosmina sp. and Chydorus sp., Copepods were
WD (cm) 16.52.9
dominated by Cyclops sp., Diaptomus sp. Furthermore,
EC (S cm-1) 523.6267.6
Brachionus sp., Keratella sp. under Rotifera and Arcella
TDS (mgl-1) 302.4543.4
sp. under Protozoa were found as dominant genera
DO (mgl-1) 6.326.5
during the study period. Relative abundance of
FCO2 (mgl-1) 12.055.4
zooplankton indicated that Cladocera (29%) was the
NO3-N (mgl-1) 1.341.2
most dominant community followed by Copepoda
PO4-P (mgl-1) 0.140.184
(24%), Protozoa (22%), Ostracoda (13%), while
TA (mgl-1) 29.479.5
Rotifera (12%) showed least dominance during the
Cl-1 (mgl-1) 36.156.5
entire study period (Figure 2). The relative abundance
Rainfall (mm) 615.9545.3
indicated that Cladocera was the most dominant as

Saikia et al., 2017
Table 2. The checklist of zooplankton from high altitude rice wetlands of Apatani Plateau during the rice growing
season of 2013-14.
Group Family Taxa
Bosminidae Bosmina sp.
Cladocera Bosminidae Bosmina cf. tripurae (Jurine 1820)
Bosminidae Bosmina longirostris (Muller, 1776)
Bosminidae Bosminopsis sp.
Chydoridae Chydorus sp.
Chydoridae Dadaya macrops (Daday, 1898)
Chydoridae Kurzia longirostris (Daday, 1898)
Chydoridae Kurzia sp.
Chydoridae Oxyurella sp.
Chydoridae Alona sp.
Chydoridae Disperalona caudata Smirnov, 1996
Chydoridae Coronatella anodonta (Daday, 1905)
Chydoridae Alona affinis (Leydig, 1860) s. lat.
Chydoridae Alona guttata Sars, 1862
Chydoridae Celsinotum macronyx (Daday, 1898)
Chydoridae Ephemeroporus barroisi Richard 1894
Chydoridae Anthalona sp.
Chydoridae Chydorus sphaericus (Muller, 1776)
Chydoridae Chydorus ventricosus Daday, 1898
Chydoridae Chydorus cf. ovalis Kurz, 1875
Chydoridae Pleuroxus denticulatus Birge, 1879
Sididae Diaphanosoma dubium (Manuilova, 1964)
Sididae Latonopsis australis Sars, 1888
Daphniidae Daphnia lumholtzi Sars, 1885
Daphniidae Ceriodaphnia cornuta Sars, 1885
Daphniidae Ceriodaphnia sp.
Daphniidae Simocephalus sp.
Daphniidae Simocephalus mixtus Sars, 1903
Moinidae Moina sp.
Moinidae Moina micrura Kurz, 1874
Moinidae Moinodaphnia sp.
Macrothidae Macrothrix sp.
Macrothidae Macrothrix spinosa King, 1853
Macrothidae Macrothrix triserialis (Brady, 1886
Copepoda Cyclopidae Cyclops sp.
Cyclopidae Mesocyclop sp.
Cyclopidae Mesocyclop edax Forbes, 1891
Diaptomidae Diaptomus sp.
Diaptomidae Limnocalanus sp.
Diaptomidae Limnocalanus macrurus Sars, 1863
Diaptomidae Bryocamptus sp.
Diaptomidae Helodiaptomus sp.
Diaptomidae Neodiaptomus sp.
Diaptomidae Nauplii of Cyclops
Diaptomidae Nauplii of Diaptomus

Table 2. continued
Group Family Taxa
Rotifera Asplanchnidae Asplanchna sp.
Asplanchnidae Asplanchna brightwellii Gosse, 1850
Brachionidae Keratella sp.
Brachionidae Keratella valga (Ehrenberg, 1834)
Brachionidae Brachionus sp.
Brachionidae Brachionus forficula Wierzejski, 1891
Brachionidae Plationus patulus (Muller 1786)
Brachionidae Platias quadricornis (Ehrenberg, 1832)
Testudinella Testudinella patina (Hermann, 1783)
Testudinella Testudinella striata (Murray, 1913)
Dicranophoridae Dicranophorus forcipatus (Muller, 1786).
Lecanidae Lecane sp.
Lecanidae Lacane ungulata (Gosse, 1887)
Lecanidae Lacane leontina (Turner, 1892)
Lecanidae Lacane bulla (Gosse, 1851)
Lecanidae Monostyla sp.
Notommatidae Eosphera sp.
Protozoa Arcellidae Arcella sp.
Difflugiidae Difflugia sp.
Centropyxidae Centropyxis sp.
Ophryoglenidae Ophryoglena sp.
Epistylidae Epistylis sp.
Ostracoda Cyprididae Cypris sp.
Table 3. Month wise diversity, dominance, evenness and richness indices of Cladocera in high altitude rice
wetlands of Apatani Plateau during the rice growing season of 2013-14
Indices April May June July August September
Diversity index 2.138 2.842 2.869 2.755 2.815 1.298
Dominance index 0.139 0.104 0.102 0.122 0.291 0.323
Evenness index 0.129 0.134 0.385 0.109 0.288 0.188
Richness index 5.672 5.865 5.233 5.025 6.707 5.346
Table 4. Month wise diversity, dominance, evenness and richness indices of Copepoda in high altitude rice
Diversity index 1.301 1.391 1.458 1.696 1.485 1.344
Dominance index 0.276 0.210 0.296 0.111 0.222 0.264
Evenness index 0.524 0.537 0.490 0.495 0.670 0.547
Richness index 1.390 1.346 1.648 1.836 2.061 1.638
Table 5. Month wise diversity, dominance, evenness and richness indices of Protozoa in high altitude rice
Diversity index 1.263 1.388 1.374 1.132 1.203 1.264
Dominance index 0.234 0.132 0.154 0.265 0.266 0.233
Evenness index 0.707 0.801 0.790 0.620 0.666 0.707
Richness index 0.854 1.559 1.176 1.259 0.926 0.842

Saikia et al., 2017
Figure 2. Relative abundance (%) among different Figure 3. Cluster analysis showing similarity of
communities of zooplankton in high altitude rice zooplankton within months in high altitude rice
wetlands of Apatani Plateau during the rice growing wetlands of Apatani Plateau during the rice growing
season of 2013-14 season of 2013-14
The monthly cluster analysis of zooplankton showed Kurasawa (1957) and Tripathi et al. (2006). The
that species compositions of April and May were zooplankton population declined at the later stage of
similar which might be because of low density of rice rice cultivation (August-September) due to strong
stems, comparatively lower water depth, low canopy predation stress by the stocked fish accompanied by
cover of rice plant, high density of primary producers growth stages of the rice plant with full grown canopy
and also the high level of nutrients. Species that limit entry of sunshine into the field that inhibits
compositions of zooplankton in June, July and August the growth of algae suspended in water. The stocked
associated in a single cluster was perhaps due to fishes of the rice fields utilize zooplankton as main
gradually increased field water level, dense canopy food resources for their growth and survival (Beisner
cover of rice, maximum number of rice stems along and Peres-Neto, 2009). This situation in turn, probably
with predatory effect of stocked fishes. However, it is led to less amount of total zooplankton at latter stages
also to be mentioned here that the species for scarcity food resources (Bahaar and Bhat, 2013).
compositions of zooplankton in September was in The findings clearly revealed that zooplankton
different cluster probably because of some community structure showed significant variability
anthropogenic activities by the farmers at harvest. within a very short period of time under wet rice agro
ecosystem. The alteration of the wet phase to dry
Community dynamics and diversity of Zooplankton phase due to seasonality seems quite favorable for re-
The mean density of zooplankton (Figure 4) was emergence of dormant zooplankton eggs inside the
highest in May (452 individuals l-1) followed by June sediment and probably was an important source for
(446 individuals l-1) and lowest in September (89 abundant zooplankton (Chittapun et al. 2009;
individuals l-1) in the entire study area. The population Rodrigues et al. 2011) in Apatani rice fields. The
density of zooplankton was high from May-July and dispersion of ectozoochory and endozoochory of
again it went down from August to till end of the aquatic birds in foraging time, rich primary
September. The study unfolded that the average productivity and available nutrient content at the
population of zooplankton was highest in June onset of the crop cycle enhanced high density of
probably because of high algal abundance zooplankton in the rice agro-ecosystem (Baltanas,
accompanied with heavy rainfall which triggered the 1992; Figuerola and Green, 2002; Figuerola et al.
growth of certain specific zooplankton genera (e.g. 2003; Lin et al. 2003). Moreover, Apatani farmers very
Bosmina sp., Cyclops sp. and Cypris sp.) in summer but frequently used to clear weeds in their wet rice fields
declined later in water recession period with the onset that allowed sufficient solar influx into field water
of winter. Similar observation was also reported by enhancing the multiplication of zooplankton.

probably minimize the turbidity of rice field water and

allowed photosynthesis for the formation of sufficient
food resources for them (Viroux, 2002; Santhanam and
Perumal, 2003; Jose and Sanalkumar, 2012). It was
found that the density of Copepods (Figure 6) were
maximum in July (129 individuals l-1) and minimum in
May (35 individuals l-1) during 2013 while in 2014,
highest (142 individuals l-1) density was in August and
lowest density was in April (19 individuals l -1).
However, the density of Protozoa (141 individuals l -1)
and Rotifera (101 individuals l-1) depicted their
dominance in May, whereas during September,
Protozoa (7 individuals l-1) and Rotifera (5 individuals
Figure 4. Monthly variations in total density of l-1) exhibited lower population density in 2013
zooplankton (individuals l-1) in high altitude rice
(Figure 7 and 8). Monthly variations of Protozoa (15
wetlands of Apatani Plateau during the rice growing
season of 2013-14. individuals l-1) and Rotifera (2 individuals l-1) follow
similar trend where their lower value was observed in
September and their higher values were in May where
The Kruskal-Wallis test confirmed that the diverse Protozoa had 100 individuals l-1 and Rotifera had 131
communities of zooplankton showed significant individuals l-1 during 2014 (Figure 7 and 8). In the
monthly variation (p<0.01). The herbivorous present findings, it was also noted that the Rotifera
zooplankton devoured by fishes regulated population and Protozoa diversity and density were high from
dynamics of algal communities through their grazing tillering to end of the transplanting stage of rice (May)
processes (Carpenter et al. 1985). It may also be and then sharply declined up to harvesting phase
explained that higher population density of suspended (September) in all the rice fields of Apatani Plateau
algae had increased the population density of which cohere with the finding of Bahaar and Bhat
zooplankton (Goldyn and Kowalczewska-Madura, (2013). The rich diversity of both Protozoa and
2008). The prey-predator interaction may be Rotifera was perhaps due to the higher population of
considered as one of the most important factors for bacteria and low water flow which caused
abundance of zooplankton which is mainly influenced decomposition of household sewages that ultimately
by algal population (Striebel et al. 2012). The algae enhanced the production rate of organic materials
which were persistent in such a situation had probably (Kumar et al. 2004; Majagi and Vijaykumar, 2009;
been fed rapidly by zooplankton during the rice Dhembare, 2011; Bhat et al. 2014, Sharma et al. 2014).
growing season (Godyn and Kowalczewska-Madura, Additionally, the sufficient sunlight also stimulates the
2008). The mean density of Cladocera were maximum reproductive strategies of Protozoa and Rotifera which
in June in 2013 (364 individuals l-1) and 2014 (473 might also be ascribed to their maximum diversity and
individuals l-1) and the minimum values of 2013 (11 density in May. Conversely, low amount of detritus
individuals l-1) and 2014 (25 individuals l-1) were in loading and increased water flow may be one of the
September (Figure 5). Besides, the Cladoceran causes behind the less population of Protozoa during
population showed increase in June probably due to the end of the rice cropping cycle (Sharma et al. 2014).
relatively high water depth (Saikia et al. 2016) which
Table 6. Month wise diversity, dominance, evenness and richness indices of Rotifera in high altitude rice wetlands
of Apatani Plateau during the rice growing season of 2013-14
Diversity index 1.389 1.533 1.426 1.024 1.427 1.040
Dominance index 0.197 0.124 0.176 0.211 0.184 0.222
Evenness index 0.802 0.926 0.942 0.830 0.833 0.557
Richness index 1.022 1.820 1.611 1.276 0.841 1.443

Saikia et al., 2017
Table 7. Month wise diversity, dominance, evenness and richness indices of Ostracoda in high altitude rice
Diversity index 0.000 0.000 0.000 0.000 0.000 0.000
Dominance index 1.000 1.000 1.000 1.000 1.000 1.000
Evenness index 1.000 1.000 1.000 1.000 1.000 1.000
Richness index 0.111 0.213 0.274 0.358 0. 696 0.341
Table 8. Month wise taxa density (individuals l-1) of Cladocera in high altitude rice wetlands of Apatani Plateau
during the rice growing season of 2013-14
Species April May June July August September
Bosmina sp. 1 4 1 0 3 2
Bosmina tripurae 4 7 59 189 9 4
Bosmina longirostris 5 17 107 30 13 5
Chydorus sp. 2 2 3 1 1 0
Dadaya macrops 1 1 14 1 1 1
Kurzia sp. 0 0 1 1 0 0
Kurzia longirostris 1 1 2 1 1 1
Oxyurella sp. 0 1 5 7 2 2
Alona sp. 0 0 2 0 1 0
Disperalona caudata 1 1 1 1 1 1
Alona affinis 0 1 4 2 1 0
Latonopsis australis 0 1 1 0 0 0
Bosminopsis sp. 1 0 4 1 1 1
Alona guttata 1 2 2 1 1 1
Coronatella anodonta 2 3 1 0 0 1
Celosinotum macronyx 0 1 2 1 1 2
Ceriodaphnia cornuta 0 1 2 1 1 0
Ephemeroporus barroisi 0 2 11 3 2 2
Chydorus sphaericus 4 5 1 1 2 1
Chydorus ventricosus 2 1 2 1 1 1
Chydorus ovalis 1 1 2 2 2 0
Pleuroxus denticulatus 0 4 4 1 1 0
Diaphanosoma dubium 1 0 3 4 1 0
Anthalona sp. 1 2 4 2 0 1
Ceriodaphnia sp. 1 6 2 1 3 0
Daphnia lumholtzi 5 8 4 3 1 1
Simocephalus sp. 1 1 3 1 0 0
Simocephalus mixtus 1 1 3 3 2 1
Moina sp. 2 2 2 1 1 1
Moina micrura 1 1 2 1 1 0
Moinodaphnia sp. 1 4 1 2 1 1
Macrothrix sp. 1 2 3 1 1 1
Macrothrix spinosa 0 2 3 1 1 1
Macrothrix triserialis 0 0 1 1 2 0

(a) 2013 (b) 2014

Figure 5. Monthly variations in density of Cladocera (individuals l -1) present in high altitude rice wetlands of
Apatani Plateau during the rice growing season of 2013-14
(a) 2013 (b) 2014

Figure 6. Monthly variations in density of Copepoda (individuals l -1) in high altitude rice wetlands of Apatani
Plateau during the rice growing season of 2013-14
(a) 2013 (b) 2013

Figure 7. Monthly variations in density of Protozoa (individuals l-1) in high altitude rice wetlands of Apatani

Saikia et al., 2017
(a) 2013 (b) 2014

Figure 8. Monthly variations in density of Rotifera (individuals l-1) in high altitude rice wetlands of Apatani
(a) 2013 (b) 2014

Figure 9. Monthly variations in density of Ostracoda (individuals l -1) in high altitude rice wetlands of Apatani
Table 9. Month wise taxa density (individuals l-1) of Copepoda in high altitude rice wetlands of Apatani Plateau
Cyclops sp. 40 16 27 36 56 20
Diaptomus sp. 15 15 26 23 35 11
Nauplli of Cyclops 13 4 5 3 3 2
Nauplli of Diaptomus 1 0 0 1 4 0
Neodiaptomus sp. 1 3 0 0 3 0
Limnocalanus sp. 3 1 1 1 5 2
Mesocyclop sp. 2 2 5 6 6 1
Bryocamptus sp. 0 0 4 2 2 1
Limnocalanus macrurus 0 0 1 3 3 2
Heloidiaptomus sp. 0 0 0 3 3 0
Mesocyclop edex 0 0 1 0 8 0

Table 10. Month wise taxa density (individuals l-1) of Protozoa in high altitude rice wetlands of Apatani Plateau
Arcella sp. 58 50 32 19 13 7
Epistylis sp. 21 19 5 3 3 1
Diflugia sp. 17 8 10 3 1 1
Centropyxis sp. 5 23 22 4 6 1
Opheroglena sp. 7 9 6 1 1 3
Table 11. Month wise taxa density (individuals l-1) of Rotifera in high altitude rice wetlands of Apatani Plateau
Keratella sp. 18 16 2 1 2 1
Brachionus sp. 18 42 16 3 4 2
Lecane sp. 3 20 2 5 1 1
Eosphera sp. 5 16 1 1 1 0
Asplanchna sp. 6 22 2 2 1 0
Asplanchna brightwelli 0 1 1 0 0 0
Keratella vulga `1 2 0 0 0 1
Brachionus forficula 1 1 1 0 0 0
Plationus patulus 0 0 0 0 1 0
Platias quadricornis 2 3 1 1 0 0
Eosphera sp. 1 1 1 0 0 0
Testudinella patina 2 2 0 0 1 1
Testudinella striata 1 3 1 1 0 0
Dicranophorus forcipotus 1 1 1 2 0 0
Lecane ungulate 2 1 1 0 0 0
Lecane leontina 5 4 3 0 1 1
Lecane bulla 0 0 0 1 0 0
Table 12 Month wise taxon density (individuals l-1) of Ostracoda in high altitude rice wetlands of Apatani Plateau
Cypris sp. 13 12 21 25 11 3
The maximum and minimum population density of The community diversity, dominance, evenness and
Ostracoda (Figure 9) was found in June (87 richness of different communities of rice fields
individuals l-1) and April, 2013 (17 individuals l-1) zooplankton presented in Table 3-Table 7. Cladocera
respectively. But in 2014 (Figure 9), peak value was showed maximum Shannon-Wiener diversity value
found in July (50 individuals l-1) and dip values were in (Table 3) in June (2.869) and the minimum was in
September (6 individuals l-1). Cladocerans might have September (1.298). Copepod (Table 4) varied from
suppressed the population density of Rotifers (Gilbert, 1.301 (April) to 1.696 (July) whereas Protozoa (1.388)
1988; Nogrady et al. 1993; Shurin et al. 2006; Hulyal and Rotifera (1.533) showed peak diversity in May
and Kaliwal, 2008). while Protozoa (1.132) and Rotifera (1.024) showed

Saikia et al., 2017
least value in July (Table 5 and 6). The evenness Computed species richness and evenness indices
indices of Cladocera (0.385) and Rotifera (0.942) were assured the congenial habitat for the existing
high in June (Table 3 and 5) whereas Cladocera zooplankton communities in the rice fields of the study
(0.109) and Protozoa (0.620) showed low evenness in area. On the other hand, the occasional low evenness
July (Table 3 and 6). The peak evenness of Copepoda of some species might have been caused due to
was in July (0.670) and the least value for evenness of frequent deweeding activities and other related
Copepoda (0.490) and Rotifera (0.557) were noticed in agronomic disturbances within the flooded rice field.
June and September respectively (Table 4 and 5). The mean density of all the zooplankton taxa
Protozoa had the high evenness (0.801) in May (Table belonging to different communities were also
6). The evenness value (1.000) of Ostracoda (Table 7) calculated (Table 8- Table 12) according to different
indicates equal distribution of the taxa in all the months during the whole study period. Simpson et al.
months. The Cladoceran (Table 3) were also having (1994) also reported similar observation that high
the high richness in August (6.707) and the low in July population of Copepods and low population of
(5.025). The richness of Copepoda (Table 4) in the Ostracods towards the end of the crop cycle. So,
wet rice fields of Apatani Plateau ranged from 1.346 predatory action of Copepods might have decreased
(May)-2.061 (August). Protozoa (Table 6) also the abundances of Rotifers in the end phase of rice
showed the high richness in May (1.559) and low in (Badsi et al. 2010). The frequent availability of
September (0.842). Similarly, the richness indices of crustaceans nauplii during the whole sampling periods
Rotifera (Table 5) were found minimum in August indicated their active reproductive phases (Sharma,
(0.841) and maximum in May (1.82) respectively. 2011; Bhat et al. 2014) in the flooded fields.
Figure 10. Linear regression between physico-chemical properties of rice field water and total density of
zooplankton in the study area

Linear regression analyses between the different such zooplankton persisted under rice canopy till
physico-chemical properties of rice field water and complete recession of the water with seasonality. All
total density of zooplankton (Figure 10) showed the agro input in this ecosystem with organic manures,
positive relation with TH, NO3-N, EC, TDS, CaH, WD decomposed rice stubbles and macrophytes along with
and DO and negative relation with WT and PO4-P. biotic and abiotic factors regulate various
Zooplankton showed a negative relation with WT, characteristics of population properties of the
NO3-N and FCO2 in the present study which also zooplankton community in rice fish system. For
corroborates with the observation of Sharma (2011); growth and development, the stocked fishes in the rice
Sharma and Sharma (2011) and Ahmad et al. (2012). A field may have utilized and assimilated available
positive relation of total density of zooplankton with aquatic biomass along with zooplankton through
TH, WD, TDS and EC have been recorded during the trophic interactions. Therefore, high altitude flooded
study period, which was also drawn by Ramakrishnan environment of Eastern Himalaya enriched with such
and Sarkar (1982); Bhati and Rana (1987); Ahmad and natural fish feeds hold sufficient scope to raise fish as
Krishnamurthy, (1990); Kumar and Datta (1994); concurrent crop of rice in the same field.
Jhingran (1997); Hujare, (2005); Ratushnyak et al.
(2006); Datta (2011); Ahmad et al. (2012). The Acknowledgments
multiple regression analysis determined the overall The first author gratefully acknowledged the
interaction of different zooplankton communities, viz., University Grants Commission (UGC), Government of
Cladocera, Copepoda, Protozoa, Rotifera and India for providing the financial support in the form of
Ostracoda with different physico-chemical properties UGC NON-NET fellowship. We also appreciate the
of the rice field water (Table 13) during the study Department of Ecology and Environmental Science,
seasons. The multiple regression analyses revealed Assam University, Silchar, Assam and Department of
that density of Cladocera was related with increase in Zoology, Rajiv Gandhi University, Rono Hills,
WD, PO4-P, TA, CaH and Cl-1 which also corroborated Arunachal Pradesh for laboratory facilities. Authors
the findings of Tidame and Shinde (2009), Ahangar et are also thankful to the Apatani farmers and Govt.
al. (2012). Copepod density was related with increase meteorological station for extending their cooperation
in EC, WD and Cl-1and decrease in DO, TH and PO4-P. during the field sampling and data collection. Thanks
Rotifera density was related with increase in FCO2, DO, are also due to Mr. Rubu Tagio, Dipankar Saikia,
TDS and TH. Protozoan density was related with Pramod Saikia and Hemanta Baruah who assisted
decrease in Cl-1 and increase in DO, TDS, NO3-N and during each of the field visit of the first author. We
FCO2. Rotifera density showed a positive correlation would also like to acknowledge our two fellow
with TH, TDS, FCO2 and DO which was agreeable to the laboratory mates for their support during the
view of Hulyal and Kaliwal (2008); Tidame and Shinde preparation of the manuscript.
(2012); Chandrasekar, (2009); Sharma, (2009); Bera
et al. (2014); Devi and Kumar (2014); Hussain et al. Conflicts of interest: The authors stated that no conflicts of
(2016). A negative relation was observed between interest.
protozoan density and Cl-1 whereas density of
Copepoda depicted positive relation with Cl -1 which
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RESEARCH ARTICLE
Anatomical markers and Phytochemical study of different plant

parts of Bacopa monnieri (L.) Wettst.
Varshney Anju 1*, Chandra Naresh2 and Patil Avinash3
1Department of Botany, 2 Principal, 3 Vice Principal, Birla College of Arts, Science and Commerce, Kalyan (W),
Maharashtra 421304 , India
*Corresponding author email: varshneyanju@yahoo.co.in

Received: 08.07.2017 Bacopa monnieri (L.) Wettst. (Scrophulariaceae) commonly known as
Accepted: 20.08.2017 brahmi, has been used in the traditional systems of medicines as "Medhya
Published : 19.09.2017 Rasayana" as it increases mental clarity and brain stimulating action. It is
also being used as anti-inflammatory, analgesic, antipyretic, sedative,
Editor:
antioxidant and anti-cancerous agent. The present study was carried out to
Dr. Arvind Chavhan
provide characteristic anatomical markers for root, stem and leaf, along
Cite this article as: with this study preliminary photochemical analysis was also carried out for
Varshney Anju, Chandra Naresh and the presence of major secondary metabolites which can act as the indicator
Patil Avinash (2017) Anatomical of bioactivity of the different plant parts of Bacopa monnieri (L.) Wettst. The
markers and Phytochemical study of distinguishing features for root and stem are simple starch grains, xylem
different plant parts of Bacopa vessel with spiral thickenings, pitted tracheids and for leaf are sub-
monnieri (L.) Wettst; International J. epidermal foliar idioblast, sessile, multicellular trichome and prismatic
of Life Sciences, 5 (3): 379-386.
calcium oxalate crystals. Presence of two types of stomata i.e. anomocytic
and diacyitc in the same field of vision under 45X magnification, are seen in
the surface preparation as well as in the powder microscopy of Bacopa
Copyright: 2017| Author (s), This monnieri leaf. This is the major observation of the study. All these
is an open access article under the distinguishing features can be used as anatomical markers for correct
terms of the Creative Commons indentification of root, stem and leaf of the plant. Preliminary
Attribution-Non-Commercial - No phytochemical analysis revealed the presence of alkaloids, saponins,
Derivs License, which permits use flavonoids, tannins, carbohydrates, proteins and anthroquinones in
and distribution in any medium, different plant parts of Bacopa monnieri while quantitative estimation
showed that alkaloids (mg /g dry weight) in root, stem and leaf were found
to be 1.67 0.577, 47.00 0.81 and 53.07 2.08 respectively; saponins (mg
adaptations are made. /g dry weight) in root, stem and leaf were 7.50 0.71, 23.33 1.53 and
57.00 2.65 respectively and flavonoids (mg /g dry weight) in stem and
leaf were 25.33 1.53 and 27.67 2.08 respectively. Thus the findings of
the present study will provide referential pharmaco - botanical information
for correct identification and authentication of the crude drug and its plant
parts.
Keywords: Bacopa monnieri (L.) Wettst., foliar idioblast, anomocytic,

diacytic types of stomata, prismatic calcium oxalate crystal and
phytoconstituents.

Varshney et al., 2017
INTRODUCTION present in the plants is of equal importance (Harborne,

2007). Phytochemicals are divided into two groups
In the last few decades there is an exponential growth (according to their functions in plant metabolism),
in the field of herbal medicine. It is getting popularize these are primary and secondary metabolites. Primary
in developing countries owing to its natural origin and metabolites are compounds like carbohydrates,
lesser side effects. Nowadays, herbal medicines are proteins, lipids which are essential for the plant
being manufactured on a large scale in mechanical metabolism. Secondary metabolites are compounds
units, where manufacturers are facing many problems which are not essential for the plant metabolism as
such as availability of good quality of raw material, such but are formed as byproducts in the biochemical
authentication of raw material, availability of pathways. These include very interesting and useful
standards, proper standardization methodology of classes of compounds like alkaloids, terpenoids,
drugs and formulations, quality control parameters etc anthocyanins, saponins phenolic compounds like
(Patil et al., 2011). For pharmacological or flavonoids, tannins, etc. These secondary metabolites
pharmaceutical use, scrutiny of a crude drug for its can be utilized for the identification of the plant
botanical identity is required (Poornima et al., 2009). material (Krishnaiah et al., 2009).
Generally herbal formulations use fresh or dried plant
parts. Correct knowledge of such crude drugs is very Bacopa monnieri (L.) Wettst., an important medicinal
important aspect in preparation, safety and efficacy of plant belongs to family Scrophulariaceae is used in the
the herbal product. Pharmacognosy is a simple and traditional systems of medicines as memory booster
reliable tool, by which complete information of the (Gohil and Patel, 2010). It is also referred to as,
crude drug can be obtained (Modi et al., 2010). Herpestis monniera L. Kunth. Bacopa monnieri (L.)
Wettst. locally known as water hyssop, brahmi or
Anatomical structure is very important for studying Jalanimba in India. The name brahmi is derived from
biological specimens for the purpose of classification, the word 'Brahma', the mythical 'creator' in the Hindu
pathological changes and the physiological significance pantheon. As the brain is the center for creative
of certain cell organelles or structures in relation to activity, any compound that improves the brain health
the habitat (Coopoosamy and Naidoo, 2011). is called brahmi, which also means 'bringing
Microscopy allows the identification of herbal drugs knowledge of the supreme reality (Prasad et al., 2008).
and the detection of individual components of a Brahmi is a small, smooth, hairless, somewhat fleshy
mixture. It is important to ensure quality and purity of and creeping herb. The plant is a short duration annual
herbal medicines in order to maximize the efficacy and herb, frequent in moist habitat and water edges
minimize adverse side effects. Adulteration and throughout tropical and subtropical India. It grows
misidentification of herbal drug can cause serious best near flowing water and wet lands in plain and
health problems to consumers (Serrano et al., 2010). foothills and is particularly abundant in monsoon.
Brahmi can grow in a variety of soil types if the habitat
Several medicinal plants have been used as a source of provides wet and semi shade conditions. Near- neutral,
many potent and powerful drugs. Medicinal property clayey loam to clayey soils are best suited for the
of a plant is due the presence of chemical entities growth of Bacopa monnieri. In North India, it can grow
(phytochemical constituents) which are extracted in a wide range of temperatures 150 C - 400 C and soil
from the plant or the plant parts (Mahesh and Satish, pH 5.0- 7.5. However, it can even grow well in soils
2008). Medicinally important plants contain diverse with pH 7.5 or even more. It becomes dormant during
group of phytochemicals with great variations in the winter months except when grown near running
solubility and stability (Mallikharjuna et al., 2007). water. Flowers and fruits appear in summer (Agro
Phytochemical progress has been aided enormously by tech, 2008).
the development of rapid and accurate methods of
screening plants for particular chemicals (Banso, The pharmacological effects of Bacopa monnieri are
2009). Aim for the phytochemical analysis is the attributed to the presence of a number of biologically
characterization of an active principle responsible for active compounds including alkaloids (nicotine and
some toxic or beneficial effect shown by a crude plant herpestine), saponins and sterols (Monograph, 2004),
extract when tested against a living system. d-mannitol, hersaponin, monnierin, betulic acid,
Quantification of the different phytoconstituents stigmastarol, beta-sitosterol, as well as numerous

Anatomical markers and Phytochemical study of different plant parts of Bacopa monnieri (L.) Wettst.
bacosides and bacopasaponins (Agro tech, 2008). The characters and preliminary phytochemical analysis of
compounds responsible for the memory enhancing different plant parts (root, stem and leaf) of Bacopa
effects of Bacopa monnieri are triterpenoid saponins monnieri (L.) Wettst.
called "Bacosides" (Srinivasa et al., 2011). The major
chemical shown to be responsible for neuropharma-
cological effects and the nootropic action or MATERIALS AND METHODS
antiamnestic effect of Bacopa monnieri was bacoside A
(Gohil and Patel., 2010). Herbarium of Bacopa monnieri (L.) Wettst. was
prepared and authenticated from Blatter Herbarium,
The whole plant of Bacopa monnieri is used as brain St. Xaviers College, Mumbai. Bacopa monnieri (L.)
tonic in indigenous system of medicines. Brahmi is Wettst. plants were collected from Keshavsrushti,
considered among one of the Celestial drugs (Divya Uttan, Bhayander, Mumbai. Root, stem and leaves were
ausadhi) when consumed with milk for 6 months. The separated, washed under running tap water and
herb is generally confused with another herb blotted dry. All the separated plant parts were dried in
Centella asiatica that is morphologically very different preset oven at 40 20C for about a week, grounded
from that of Brahmi. The juice of the boiled whole into powder and used for further analysis.
plant is given to children for relief in bronchitis and
diarrhoea. The paste of the leaves is used as a remedy Anatomical Study
for rheumatism. Ghee fried leaves are given in Fresh plant parts were used for morphological and
hoarseness complaints. The leaves and tender stalks microscopical studies. Surface preparations were used
are reported to be eaten in the west Bengal. Its juice to study type and structure of stomata and trichomes
along with ginger juice, sugar and bark ext. of and stomatal index was also calculated. The fine
Moringa oleifera is given to children in stomach sections were stained by safranine and mounted in
disorders. Decoction of leaves is given in cough. glycerine, were photographed under light microscope
Brahmi Ghrita a medicated ghee of brahmi when using 45X magnifications.
given in a dose of 1-2 tolas along with milk twice a
day remove insanity problems. Brahmi Rasayana Phytochemical Analysis
imporves memory power when taken in a dose of 1-3 Preliminary phytochemical screening of different plant
mashas along with honey and ghee (Krishnamurthy). parts (root, stem and leaf) of Bacopa monnieri (L.)
Paste or juice of fleshy leaf and stem of brahmi were Wettst. was carried out by using standard methods .
mixed with sugar, jaggery or honey to make it more Aqueous and ethanolic extracts of root, stem and leaf
palatable due to its bitter taste. Some of the known were used for qualitative analysis. Quantitaive
preparations with brahmi are Sarasvatarishta (a estimation of secondary metabolites such as alkaloids,
decoction used as a brain tonic), Brahmi Rasayana (a saponins and flavonoids in different plant parts were
rejuvenating formulations with other herbs), Brahmi also carried out by using standard methods (Harborne,
Taila (medicated oil) and Brahmi Sarbat (a cooling 1973; Khan, 2001; Boham and Kocipai method, 1994
drink) (Prasad et al., 2008). repectively).
Brahmi has been used in Ayurvedic formulations for

conditions ranging from catarrhal complaints,
gastrointestinal disturbances due to excessive RESULTS AND DISCUSSION
tobacco use, habitual abortions and high blood sugars
due to anxiety disorders. In certain parts of India, Anatomical Study
brahmi is believed to be an aphrodisiac; in Sri Lanka,
under the name of Loonooweella, brahmi is Morphological Study
prescribed for fevers; in the Philippines, it is used as a The morphological characters can be diagnostic
diuretic (Prasad et al., 2008; Russo and Borrelli, parameters for the plant.
2005).
Roots: Creamish yellow in colour, thin tapering, wiry,
The present study included examination of small, branched and arising from the nodal region of
morphological and microscopical characters, powder the stem.

Figure 1: (A) Morphology of whole plant; (B) Stem with node and internode; (C) Leaves; (D) T.S. of root;
(E) T.S. of stem; (F) to (G) T.S. of leaf; (H) to (J) Enlarged surface view of leaf,
Keywords: E= Epidermis, Co= Cortex, Ar = Aerenchyma, Ed= Endodermis, Sg = Starch grains, Gl tr = Glandular
trichome, Id= Idioblast, Caox cr = Calcium oxalate crystal, VB = Vascular bundle, Xy = Xylem, Ph = Phloem, Msl =
Mesophyll, Dcy st = Diacytic stomata and Anmcy st = Anomocytic stomata.

Stem: Green or purplish-green in colour, prostrate, were comparatively larger than the cells of lower
thick and fleshy, herbaceous, soft, with prominent epidermis and covered with striated cuticle. Presence
nodes and internodes Figure 1 A& B]. of sub - epidermal foliar idioblasts (found in the form
of empty cavity) and a centrally located conjoint,
Leaves: Simple, sessile, glabrous, opposite and collateral vascular bundle encircled by a
decussate, obovate -oblong to spathulate in shape, parenchymatous sheath were observed [Figure 1 F].
apex is obtuse and margins are completely entire, 1-3 Few crystals of calcium oxalate were seen embedded
nerved or penninerved, punctate, faint green in colour in the undifferentiated mesophyll tissue [Figure 1G].
Figure 1 A, B & C].
Powder characteristics
Flowers: White with violet and green bands and
spangled with shining dots while fresh, short lived and Whole plant powder showed presence of simple,
colour lightens gradually, actinomorphic, solitary, round to oval shaped starch grains, anomocytic and
axillary, bracteate, bracteoles are shorter than pedicel, diacytic types of stomata, prismatic calcium oxalate
pedicel is slender in shape [Figure 1 A, B & C]. crystals, xylem vessels with spiral thickenings and
Microscopical Study pitted tracheids [Figure 2 A1, A2, A3 and A4].
Transverse section (T.S.) of root showed single Root powder is brown in colour. The various
layered epidermis with wide cortical arenchymatous diagnostic characteristics of root powder are shown in
region. Endodermis was distinct and single layered Figure 2 B1, B2, B3 and these are simple starch grains,
while pericycle was not differentiable. Central region xylem vessels with spiral thickenings and pitted xylem
was occupied by stele consisted of 1-5 layers of tracheids.
peripheral phloem and centrally located xylem vessels
[Figure 1D]. Stem powder is light brown in colour and bitter in
taste. The various diagnostic characteristics of stem
Transverse section (T.S.) of stem showed single powder observed are xylem vessels with spiral
layer of epidermis, cortex with chlorenchymatous thickenings, pitted xylem tracheids, simple, round and
aerenchyma or air spaces, cortical cells with starch oval starch grains. Separate spiral rings were also
grains, single layered endodermis, 1-2 layered observed [Figure 2 C1, C2, C3 and C4].
pericycle, continuous vascular ring composed of a
narrow zone of phloem towards periphery and a wide Leaf powder is green in colour and bitter in taste. The
ring of xylem towards centre, centrally located various diagnostic characteristics of leaf powder are
parenchymatous pith with simple, round to oval starch the presence of anomocytic and diacytic types of
grains [Figure1 E]. stomata, prismatic calcium oxalate crystals in
mesophyll tissue and simple starch grains [Figure 2 D1,
Surface preparation of leaf showed the presence of D2, D3 and D4]. Quantitative determination of leaf
two types of stomata i.e. anomocytic and diacytic content included number of both types of stomata and
[Figure1J], 8- celled sessile glandular trichomes stomatal index was done with the help of standard
[Figure1 H & I]. The anomocytic types of stomata were methods and results are tabulated in Table 1.
more in number in comparison to diacytic type.
Phytochemical analysis
Transverse section (T.S.) of leaf showed distinct
upper and lower epidermis, cells of upper epidermis Preliminary phytochemical analysis
The results of preliminary phytochemical analysis are
Table 1: Quantitative leaf microscopy of Bacopa tabulated in Table 2. The phytochemical analysis
monnieri (L.) Wettst. revealed the presence of different chemical
Parameter Value compounds such as carbohydrates, alkaloids, saponins,
Stomatal index 10.21% flavonoids, tannins and anthroquinones. Among the
Stomata number 34 different plant parts of Bacopa monnieri (L.) Wettst.,
Number of anomocytic stomata 23 leaves were found to contain higher concentrations of
Number of diacytic stomata 11 phytoconstituents (alkaloids, saponins and flavonoids)

followed by stem and root.The amount of alkaloids, amount of saponins (mg /g dry weight) were found to
saponins and flavonoids in different plant parts are be 57.00 2.65 in leaves, 23.33 1.53 in stem and 7.50
described in Table 3.. The amount of alkaloids (mg /g 0.71 in root while flavonoids (mg /g dry weight)
dry weight) were found to be 53.07 2.08 in leaves, were found to be 27.67 2.08 in leaves and 25.33
47.00 0.81 in stem and 1.67 0.577 in root. The 1.53 in stem.
Figure 2: Powder characteristics of Bacopa monnieri (L) Wettst.; A1 to A4 whole plant; B1 to B3 root; C1 to C4
stem; D1 to D4 leaf
Keywords: Sg= Starch grains; Ec= Epidermal cell; Pc= Parenchymatous cells; Msl=Mesophyll; Pt = Pitted tracheid;
Sp = Spiral thickening; Caox cr = Calcium oxalate crystal; Dcy st =Diacytic stomata; Anmcy st = Anomocytic
stomata.

Table 2: Phytochemical constituents in different plant parts (root, stem & leaf) of Bacopa monnieri (L.) Wettst.
Tests Name of the Root Stem Leaf
Method /test WE EE WE EE WE EE
Carbohydrates Anthrone test - + + + + +
Proteins Folin Lowry test - + - + - +
Alkaloids Harbornes method - + - + - +
Saponins Harbornes Froth and
+ + + + + +
Emulsion tests
Flavonoids Harbornes method - + + + + +
Tannins Harbornes method - + + + + +
Terpenes and steroids Harbornes method - - - - - -
Glycosides Harbornes method - - - - - -
Anthroquinone Harbornes method - + + + + +
Keys: WE = Water extract, EE = Ethanolic extract, + = Present, - = Absent
Table 3: Quantitative analysis of Bacopa monnieri (L.) Wettst. different plant parts
Phytoconstituents Different plant parts
(mg / g dry wet.) Root Stem Leaf
Alkaloids 1.67 0.577 47.00 0.81 53.67 2.08
Saponins 7.50 0.71 23.33 1.53 57.0 2.65
Flavonoids 0.0 25.33 1.53 27.67 2.08
All values are mean of three determinants and expressed as Mean S.D.
CONCLUSION under 45 X magnification of light microscope and this

is the major finding. Anomocytic type of stomata were
Bacopa monnieri (L.) Wettst. is a popular medicinal observed more in number in comparison to diacytic
plant, which is being used for various medicinal stomata. The stomatal index and number of
properties. Microscopic identification of medicinal anomocytic and diacytic types of stomata were found
plant is essential to distinguish any adulterants or to be 10.21%, 23 and 11 respectively. Presence of two
other similar species in the crude herbal mixtures. types of stomata in a single plant may be the result of
Microscopic characteristic findings of different plant evolutionary changes or environmental changes.
parts of Bacopa monnieri may be used as anatomical
markers to identify the plant and its plant parts. The Another distinguishing feature of the Bacopa leaf was
anatomical markers for root are presence of simple the presence of sub- epidermal foliar idioblasts. These
starch grains, xylem vessels with spiral thickenings were in the form of air spaces. Idioblasts are isolated
and pitted tracheids; for stem are presence of xylem plant cells which differ from neighbouring cells or
vessels with spiral thickenings, pitted xylem tracheids, tissues and contain non-living substances such as oil,
simple, round and oval starch grains and for leaf are latex, gum, resin, tannin or pigments etc. Some can
presence of anomocytic and diacytic types of stomata, contain mineral crystals such as calcium oxalate or
upper and lower sub - epidermal idioblasts, sessile, carbonate or silica. They have various functions such
multicellular glandular trichome, prismatic calcium as storage of reserves, excretory materials, pigments
oxalate crystals, simple and round starch grains. All and minerals. Karrfalt and Tomb (1983) described
the above mentioned anatomical markers were also idioblast as air spaces which were irregular in shape
observed in whole plant powder of Bacopa monnieri and lacked an epithelium. Presence of foliar idioblasts
(L.) Wettst. in the form of air spaces were also reported in the
other members of family Scrophulariaceae, such as
In the present study, two types of stomata i.e. Scrophularia and Verbascum genera (Lersten and
anomocytic and diacytic in a same field of vision have Curtis, 1997).
been observed in Bacopa monnieri (L.) Wettst. leaf,

These anatomical markers can be used in Krishnamurthy KH. Traditional Family Medicine. Isabgol,
authentication and for quality assurance of Bacopa Gokhru and Brahmi. Health Series, 56-75.
monnieri for Herbal medicine manufacturing Lersten RN and Curtis DJ. (1997) Anatomy and distribution
of foliar idioblasts in Scrophularia and Vervsscum
industries. Results obtained from phytochemical (Scrophulariaceae). American Journal of Botany, 84(12):
analysis could make the plant useful for treating 1638-1645.
different diseases and having a potential of providing Mallikharjuna PB, Rajanna NL, Seetharam NY and
useful drugs for human use. All these findings can also Sharanabasappa, KG (2007) Phytochemical Studies of
Strychnos potatorum L.f. - A Medicinal Plant. E- Journal
serve as an important source of information to
of Chemistry, 4 (4): 510-518.
ascertain the identity and to determine the presence of
Mahesh, B and Satish, S. (2008) Antimicrobial activity of
adulterants from the raw material of Bacopa monnieri some important Medicinal Plant against plant and
(L.) Wettst. as well as plant parts used in it. human pathogens. World Journal of Agricultural
Sciences, 4 (S): 839-843.
Modi DC, Patel JK, Shah BN and Nayak BS(2010)
Pharmacognostic studies of the seed of Syzygium cumini
Acknowledgement
Linn. International Journal of Pharmaceutical Sciences,
The authors would like to thank Mrs. U.C. Bapat, Director, 1(1): 20- 26.
Blatter Herbarium, St. Xaviers College, Mumbai, to
Monograph (2004) Bacopa monnieri. Alternative Medicine
authenticate the plant material for this research work. The Review, 9(1): 79-84.
authors would also like to thank Dr. Jossy Varghese, Sr. Vice
Patil AG, Joshi KA, Patil DA and Naresh Chandra (2011)
Principal and Dr. (Mrs.) A.V. Pathak, Ex-Vice Principal, Birla Pharmacognostical standardization and HPTLC
College, Kalyan for providing the facilities for research, fingerprint of Cardiospermum halicacum L. stem.
technical advice and reviewing the manuscript. The authors Research Journal of Pharmaceutical, Biological and
would also like to thank Dr. (Mrs.) Geetha Menon, Vice Chemical Sciences, 2(2): 343- 352.
Principal, R.K. Talreja College, Ulhasanagar for valuable Poornima N, Umarajan KM and Babu K (2009) Studies on
support. anatomical and phytochemical analysis of Oxystelma
esculentum (L.f.) R.br. Ex Schltes. Botany Research
International, 2(4):239-243.
Conflicts of interest: The authors stated that no conflicts of Prasad R, Bagde US, Puspangadan P and Varma A (2008)
Bacopa monnieri (L.): Pharmacoloigcal aspects and case
interest.
study involving Piriformospora indica. International
Journal of Integrative Biology, 3(2): 100-110.
Russo A and Borrelli F. (2005) Bacopa monnieri, a reputed
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RESEARCH ARTICLE
Lichens: A myriad hue of Bioresources with medicinal properties
Ramya K and Thirunalasundari Thiyagarajan
Department of Industrial Biotechnology, Bharathidasan University, Tiruchirappalli - 620 024, Tamil Nadu
E-mail: drttns@gmail.com

Received: 12.07.2017 Lichens form an ideal model to study its bioactive metabolites. A symbiotic
Accepted: 08.08.2017 association between fungus and a chlorophyll containing partner; the green
Published : 19.09.2017 algae or cyanobacteria, and sometimes both together form lichens. Lichens
are important species for several reasons. The unique and biologically
Editor:
active substances present in the lichen thalli contribute to several
Dr. Arvind Chavhan
important medicinal values of lichens like antiviral, antibacterial, antifungal,
Cite this article as: antioxidant, cytotoxic effect and many more. All these manifold activities of
Ramya K and Thirunalasundari lichen metabolites represent the therapeutic potential of lichens that has a
Thiyagarajan (2017) Lichens: A great impact in Pharma industries. The present article is a review on the
myriad hue of Bioresources with importance of lichens in contributing towards valuable resources
medicinal properties; International J. emphasising on its medicinal properties.
of Life Sciences, 5 (3): 387-393.
Keywords: Lichens, Biomonitors, Medicinal properties, Therapeutic
potential.
Attribution-Non-Commercial - No INTRODUCTION
Lichens, the symbiosis between a fungal and algal partner, are found all
around the world. They grow in the residual areas of any extremities where
life can survive, which may be harsh for any other organisms to exist. The
adaptations are made. usual habitats of lichens are bare rock, sand, soil, dead wood, living bark
and rusty materials surviving extreme conditions of temperature. The
adaptations and survival factor in such environments has led lichens giving
rise to a large number of unique secondary metabolites. Over 20000 species
of lichens are known worldwide. While the ethnic uses of lichens and the
pharmaceutical potential of their secondary metabolites have been
recognized, the knowledge regarding a large number of lichen substances
still remains a mystery. The area of bioactivity research towards focussing
lichen derivatives as drug targets and its screening needs an emphasis. An
updated overview on the knowledge of secondary metabolites from
lichens can highlight their potentials for pharmaceuticals as well as other
applications. This review focuses on the lichen medicinal values
emphasising on its bioactive potentials. There is a need for expanding
research in this area of study exploring the unique compounds of lichens
which are promising reservoirs with high medicinal values.

Ramya K and Thirunalasundari, 2017
Lichen habitat and distribution Squamulose lichens: This form of lichen is neither a
Lichens colonize some of the most inhospitable crustose nor foliose type but an intermediate form.
habitats on earth. Lichens are known to pioneer Minute lobes are present on the dorsiventral surface
different types of hostile environments from extreme of the lichen with either presence or absence of
temperatures to desiccated colonisations. Very old rhizines.
environments as well as arid and semi-arid regions
like cryptogamic soil crusts are also conditions Lichen metabolites
favouring lichen habitat. Fog, dew or high air humidity Being one of the slowest growing symbiotic
ensures survival in dry areas (Beckett et al., 2008). associations, lichens occupy low-resource habitats and
Lichens are also reported to grow in desert regions thereby produce certain defence chemicals to survive
due to its capability to become hydrated without being in the harsh environments. These chemicals are the
in contact to any liquid or water (Printzen et al., 2012). reservoirs of lichens; the secondary metabolites that
This suggests that lichens are slow-growing as well as render chemical defence and protection against
long-living host organisms. The favourable various microbes, animal predators and plants, to
environments for lichen habitat are considered to be avoid environmental stress with respect to UV
microsites which are rocks, barren soil, man-made damage, desiccation and also for the physiological
substrates and epiphytic life-styles. Around 8% of the regulation of metabolism (Huneck & Yoshimura,
earth land surface is covered by lichens (Ahmadjian, 1996). The lichen compounds exhibit significant
1995). Lichen flora is approximately estimated to biological activities (Huneck, 1999). Lichens and their
cover 18500 species worldwide (Feuerer & metabolites have a manifold activity exhibiting
Hawksworth, 2007). antiviral, antibacterial, antifungal, antibiotic,
antitumor, allergenic, plant growth inhibitory,
Growth forms antiherbivore, antioxidant and enzyme inhibitory
Major ecological groups of lichens include: activities (Karagoz et al., 2009) (Ramya &
Epiphytic lichens which are i) corticolous found on Thirunalasundari, 2013). Hence, potential value of
the bark, ii) lignicolous found on wood and iii) these metabolites for medicinal purposes is generating
folicolous found on leaves an increasing interest in research areas.
Terricolous lichens found on ground (soil)
Muscicolous found on moss More than a thousand primary and secondary
Saxicolous found on rocks. metabolites with identified structures have been
identified in lichens (Molnr and Farkas, 2010).
Lichens have a variety of different growth forms which Primary metabolites are proteins, lipids,
are carbohydrates, and other organic compounds that are
Crustose (or Crustaceous) lichens: These types of essential to the lichens metabolism and structure
lichens encrust and spread over and into the surface which are produced by lichens fungal partner and
of their habitat which cannot be removed from the algal or cyanobacterial partners. Primary lichen
surface without crumbling away. substances are intracellularly synthesised. These
Foliose lichens: Leaf like lobes spreads over the metabolites are independently synthesised by both the
surfaces which are attached by root-like threads and symbionts having structural functions and play
can be removed with the help of a knife. essential roles in cellular metabolism. Chitin, lichenin,
Fruticose lichens: Shrubby forms with many isolichenin, hemicellulose, pectins, disaccharides,
branches and they can be removed from the surface polyalcohols, amino acids, enzymes, and pigments like
by hand. algal chromophores: chlorophyll and, -carotenes,
xanthophylls, etc., are some of the primary metabolites
Other intermediate categories include: reported in lichens (Podterob, 2008).
Leprose lichens: The surface of the lichen is
powdery or granular. There is no prominent thallus Several studies have confirmed that the secondary
and hence strongly attached with the substratum. metabolites are exclusively from fungal origin
Placodioid lichens: The thallus of the lichen is closely (Culberson & Armaleo, 1992; Hamada et al., 1996;
attached to the substratum at centrally and free at Stocker-Wrgtter & Elix, 2002). It has been reported
the margin as it lacks rhizines. by several studies that the metabolic interaction

between the fungal and its algal partner is the main can be unravelled for its unique metabolites and has
reason for the production of these secondary been the subject of many research teams. Lichen
chemicals (Molina et al, 2003). Generally, the lichen secondary metabolites are considered as potential
secondary compounds are categorised as depsides, resources as these compounds function as chemical
depsidones, dibenzofurans and pulvinic acid defence against biotic and abiotic stressed conditions
derivatives having low molecular weight (Turk et al., (Balaji & Hariharan, 2007). The name Lichen itself
2003), which are produced by the fungus alone and indicates the medicinal importance of the species
secreted onto the surface of hyphae either in which can be traced back to the Greek word, Leprous
amorphous forms or as crystals (Elix, 1996). These used for treating diseases (Llano, 1944). Lichen
substances are stress compounds or organic substances are unique to lichen symbiosis which helps
substances produced under extreme conditions. the organism to survive in extreme environmental
Phytochemical analysis of lichens has portrayed the conditions. Almost majority group of lichens have been
presence of flavanoids, saponins, alkaloids, tannins, reported to contain antibiotic property (Sharnoff,
phlobotannins, steroids and glycosides (Ramya & 1997). Among them, the best compound that has been
Thirunalasundari, 2014). demonstrated to have a wide spectrum of antibiotic
activity is Usnic acid (Rowe et al. 1991). A great
These secondary metabolites which are called the diversity of biological effects are exhibited by lichen
lichen substances have evolved through various metabolites like antiviral, antibacterial, antibiotic,
transformations leading to important biosynthetic antiinflammatory, analgesic, antipyretic,
pathways and thereby producing a complex arrays of antiproliferative and cytotoxic activities (Boustie &
compounds. Most classes of the lichen metabolites are Grube, 2005).
from the acetyl-polymalonyl pathway. Pulvinic acids
and terphenyl-quinones belong to shikimic acid Research today is focusing towards break through
derivatives while mevalonate pathway accounts for discovery of newer therapies and compounds for
the wealth of di- and triterpenoids as well as steroids better treatment of diseases without any side effects.
found in lichens (Elix et al, 2008). Though a large In this effort, hunt for compounds from natural
number of research works have reported the isolation products especially in the treatment of cancer have
and structural characterization of individual lichen been developed and commercialized. However, the
compounds, lacunae in studying the lichen metabolites challenge continues towards a search for improved
as strong therapeutics still persist. cytotoxic agents as anti-cancer drugs. Lichens are one
among the bio-resources that has many untapped
Medicinal value of Lichens metabolites which can contribute to modern day
In the thrust to discover various drug therapies, medicine.
natural products are taking a central place. Though
several thousand compounds have been identified and A brief summary on the importance of the
purified from various natural sources including plants, anticancerous property of the lichens based on recent
microorganisms, fungi etc. to treat various diseases, articles is given in Table 1.
there are many more untapped reservoirs having
enormous medicinal properties and lichens being one Lichens as traditional medicine
among them. Also due to the increase in multi drug-
resistance among pathogens, there is a growing Various species of lichen forming fungi have wide
demand for novel bioactive compounds. In the present geographical distributions which are used in the
scenario of life style, usage of synthetic drugs traditional system of medicine. Lichens are being used
continuously has developed resistance characteristics. as traditional medicines across the world. Applications
Hence, a need to find new promising molecules of of lichens emphasising on the importance of its
natural origin in the control and prevention of various medicinal values can be linked with folklore. For
human, animal and plant diseases persists. Also due to centuries, lichens have been used as ingredients in folk
long-term usage of the synthetic drugs, numerous side medicine. Also, lichens+ have been used among many
effects are being identified apart from resistance. In cultures to treat variety of ailments as part of their
search of new bioactive molecules of natural origin, traditional medicines (Dayan & Romagni, 2001).
lichens are one among the promising resources which

Table 1 : Summary on the importance of the anticancerous property of the lichens

Name of the lichen Lichen metabolite Medicinal property reported Reference
Pseudevernia Olivetoric and Has cytotoxic and oxidative effects on Emsen et al.
furfuracea (L.) physodic acid cultured human amnion fibroblasts (2017)
- Usnic acid and Strong inhibitory effects on cancer cell Galanty et al.
atranorin proliferation and migration. (2017)
Anti-invasive effects on human prostate
and melanoma cancer cells
- Physodic acid Induces ROS production and apoptosis in Cardile et al.
melanoma cells. (2017)
Evernia prunastri Evernic and usnic The metabolites have been reported as Fernndez-
Usnea ghattensis acids protective against redox impairment Moriano et al.
mediated cytotoxicity (2017)
Hypogymnia Physodic acid Cytotoxic activity observed against breast Studziska-Sroka
physodes cancer cell lines : MDA-MB-231, MCF-7, et al. (2016)
T-47D
Cetraria islandica Fumarprotocetraric Neuroprotective activity against Fernndez-
acid hydrogen peroxide-induced oxidative Moriano et al.
Vulpicida Canadensis Usnic, pinastric and stress model in astrocytes. Cytotoxic (2015)
vulpinic acids potential against human HepG2 and MCF-
7 cell lines.
Ramalina fraxinea Evernic acid, obtusatic All the samples exhibited strong Ristic et al.
and Ramalina acid, sekikaic acid anticancer activity against human (2016)
fastigiata atranorin, epithelial carcinoma (HeLa), human lung
protocetraric acid and carcinoma (A549) and human colon
usnic acid carcinoma (LS174) cells
Usnea barbata Usnic acid Anticancer and antioxidant activity Zugic et al. (2016)
Usnea strigosa Norstictic acid Inhibits breast cancer cell proliferation, Ebrahim et al.
(Ach.) migration, invasion (2016)
Pseudevernia Olivetoric acid (OLA) In-vitro antitumor activities on Emsen et al.
furfuracea and physodic acid glioblastoma cells (2016)
(PHA)
Rhizoplaca Psoromic acid In-vitro antitumor activities on Emsen et al.
melanophthalma glioblastoma cells (2016)
Xanthoparmelia Usnic, salazinic, Anticancerous activity against against Shrestha et al.
chlorochroa constictic, and Burkitt's lymphoma (Raji) cells. (2015)
norstictic acids Extracts from both lichens decreased
Tuckermannopsis Protolichesterinic acid proliferation, accumulated cells at the G0
ciliaris /G1 stage, and caused apoptosis in a
dose-dependent manner. Both lichen
extracts also caused upregulation of p53.
Parmelia sulcata Not mentioned Reports caspase-independent apoptotic Ari et al. (2015)
cell death at lower doses and genotoxic at
higher doses
Parmotrema Catechin, purpurin, An antioxidant extract induces cell cycle Ghate et al.
reticulatum tannic acid and arrest and apoptosis in MCF-7. Both (2013)
reserpine anticancerus and antioxidant acivity
reported
- Usnic acid Cell cycle arrest and cell death via Singh et al. (2013)
mitochondrial membrane. Induction of
apoptosis in human lung carcinoma cells.
Parmelia caperata Protocetraric and Strong antioxidant, antimicrobial, Manojlovi et al.
usnic acids and anticancer effects (2012)
P. saxatilis and Salazinic acid
P. Sulcata

Xanthoria parietina Parietin Cytotoxicity against Human ovarian Backorov et al.

carcinoma A2780, Human breast (2011)
Umbilicaria Hirsuta Gyrophoric acid adenocarcinoma MCF-7, Human colon
adenocarcinoma HT-29,
Human promyelocytic
Leukemia, Human T cells
lymphocyte leukemia, Jurkat Human
cervix adenocarcinoma, Human breast
adenocarcinoma,
Human colon
carcinoma
Cladonia Usnic acid Effective inhibitors of DNA synthesis Einarsdttir et al.,
Arbuscula against Breast cancer cell line - T47D and (2010)
AlectoriaOchroleuca Usnic acid Pancreatic cancer cell line
Capan-2
Knowledge of these medicinal uses is available to us (Conti et al., 2001). Accumulated pollutants have
because of the contributions of traditional knowledge shown a close correlation with the atmospheric levels
holders among various cultures. The medicinal use of of the pollutants and have proved the lichens
lichens can be traced back to the 18 th dynasty (1700- capability as an effective biomonitor (Godinho et al.,
1800 BC) where the species of Evernia and 2008). This highlights the ecological importance of
Parmeliaceae was first used as a drug (Launert, 1981). lichen species. It is observed that crustose lichens are
Lichens in traditional medicine are most commonly highly resistant to air pollution than fruticose lichens
used for treating wounds, skin disorders, respiratory which justifies its presence in majority of the polluted
and digestive issues, obstetric and gynaecologic land surfaces.
concerns (Crawford, 2007)
Lichen genomics
Medicinal uses of lichens are reported among the With the advent of the genomic era, lichenology also
cultures in Africa, Europe, Asia, North America, and steps ahead towards a focus on the molecular
South America. The lichen genera that were reported mechanics of lichenization. The importance of
to be used in traditional medicine are Usnia, Evernia, culturing lichen mycobionts and photobionts has led to
Letharia, Parmotrema, Everniastrum, Xanthoparmelia, the investigation of fungal genome sequencing.
Cladonia, Cladina, Ramalina, Lobaria, Peltigera and Genomics has paved way to explore the impossible
Umbilicaria (Crawford, 2007). The medicinal aspects of lichen genomes. Metagenomic analysis using
properties of the lichen species changes based on the taxon-specific primers is of great help in identifying
substrate of its growth which imbibes the medicinal the symbiotic partners and the biosynthetic pathways
properties. associated with the metabolites of lichens (Annette
Kampa et al., 2013). Hence, the introduction of new
Lichens as Biomonitors technologies has bypassed many of the difficulties in
Bioindicators or Biomonitors are organisms that react identifying the structural small molecules that was
to environmental pollution with their life functions previously difficult.
providing quantitative information about the
environment surrounding them. Lichens are amongst Molecular research on lichen symbiosis has been
the most sensitive organisms to environmental conducted within a phylogenetic context aiming to
changes at the ecosystem level playing a beneficial role establish an evolutionary framework, and also to gain
complementary to chemico-physical monitoring information on symbiont specificity (De Priest, 2004).
methods and thereby can be used as biomonitoring Hence, with the advent of modern techniques, strategy
systems. Some lichens absorb many toxic substances based on bioinformatic prediction, genomic techniques
like sulphur dioxide, fluorides and heavy metals from and advanced analytics can help to characterize the
the atmosphere. They readily accumulate pollutants in molecules present in lichens taking it a step forward as
their thallus like heavy metals and other components druggable molecules.
from the ambient air through their entire surface

Future scope of Lichenology Balaji P, Hariharan GN (2007) In vitro antimicrobial activity

Though industrial production of lichen metabolites is of Parmotrema praesorediosum thallus extracts. Res. J.
Bot. 2(1), 54-59.
yet to progress, approaches towards axenic cultivation
Beckett RP, Kranner I & Minibayeva FV (2008) Stress
of the lichen mycobionts have been successfully physiology and the symbiosis. In: Nash TH (ed.), Lichen
performed to analyse the secondary metabolites biology, 2nd edn. Cambridge University Press,
produced by the lichen forming fungi (Stocker- Cambridge, pp. 134-151.
Wrgtter, 2008). This has given hope in identifying Behera BC, Verma N, Sonone A, Makhija U (2005)
Antioxidant and antibacterial activities of lichen Usnea
novel molecules that can be of pharmaceutical
ghattensis in vitro. Biotechnol Lett. 2005; 27:991995.
importance. Lichens grow very slowly and hence the
Boustie JL, Grube M (2005) Lichens- a promising source of
optimization of its culturing conditions in the bioactive secondary metabolites. Plant. Genet. Res., 3,
laboratory towards obtaining abundant biomass 273-87.
production is a challenging task (Behera et al., 2009). Cardile V, Graziano AC, Avola R, Piovano M, Russo A (2017)
Once this can be solved, access to lichen-derived Potential anticancer activity of lichen secondary
metabolite physodic acid. Chem Biol Interact. 2017 Feb
substances for possible applications in pharmaceutical 1; 263: 36-45.
research can surely be a break through phenomenon Christian Printzen, Fernando Fernndez-Mendoza, Lucia
Muggia, Gabriele Berg and Martin Grube (2012)
Alphaproteobacterial communities in geographically
CONCLUSION distant populations of the lichen Cetraria aculeate. FEMS
Microbiology Ecology, 82 (2)
Conti ME, Cecchetti G (2001) Biological monitoring: lichens
Lichens are an untapped source of biological activities as bioindicators of air pollution assessmenta
of industrial importance. Their potential is yet to be review. Environ Pollut; 114:471492.
fully discovered and utilized. Lichen-derived bioactive Crawford S (2007) Ethnolichenology of Bryoria fremontii:
compounds hold great promise for biopharmaceutical wisdom of elders, population ecology, and nutritional
chemistry. M.Sc. thesis, University of Victoria, Canada.
applications towards development of new
Culberson CF, Armaleo D (1992) Induction of a complete
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secondary product pathway in a cultured lichen fungus.
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lichenoric compounds will be a continual process in of pesticides. Pesticide Outlook 12, 229-232.
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Conflicts of interest: The authors stated that no conflicts of Ebrahim HY, Elsayed HE, Mohyeldin MM, Akl MR,
Bhattacharjee J, Egbert S, El Sayed KA (2016) Norstictic
interest.
Acid Inhibits Breast Cancer Cell Proliferation, Migration,
Invasion, and In Vivo Invasive Growth Through
Targeting C-Met. Phytother Res. ,30 (4)
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RESEARCH ARTICLE
Antifungal activity of extracts of weed biomass against

Aspergillus flavus Link ex Fr., a causal agent
of yellow mold of Groundnut
Deshmukh RS
Department of Botany, B. Raghunath College, Parbhani - 431 401

Email: rsdeshmukh19@gmail.com

Received: 03.08.2017 During the present studies, weeds found in different crop fields were
Accepted: 15.09.2017 screened to observe their antifungal activity against yellow mold causing
Published : 23.09.2017 fungi of groundnut i.e. Aspergillus flavus Link ex Fr. To study the antifungal
mechanism of weed extract, poisoned food technique was followed. The
Editor:
efficacy of weed extracts was expressed as percent inhibition of mycelial
Dr. Arvind Chavhan
growth over control. Among these weeds, leaf and stem extracts of
Cite this article as: Parthenium hysterophorus L. and Commelina benghalensis L. were found to
Deshmukh RS (2017) Antifungal be most effective against yellow mold causing fungi of groundnut i.e.
activity of extracts of weed biomass Aspergillus flavus Link ex Fr.
against Aspergillus flavus Link ex Fr.,
a causal agent of yellow mold of Key words: Weed, Aspergillus flavus, Yellow mold.
Groundnut; International J. of Life
Sciences, 5 (3): 394-398.
INTRODUCTION
is an open access article under the Groundnut (Arachis hypogaea L.) is considered as most important crop in
terms of the Creative Commons the World (13th) and is most important source of edible oil (4th). Similarly
Groundnut is ranked as 3rd most important source of vegetable protein in
the world (Smith, 2002). It is recognized as a palatable poor mans nut and
is eaten and relished by all classes of people.
commercial and no modifications or In India, diseases are one of the major constraints responsible for low yield
adaptations are made. of Groundnut. Groundnut is attacked by over 55 pathogens including
viruses. The important one among them responsible for huge losses in yield
are early and late leaf spots, rusts, necrosis, bud blight, collar rot, stem rot,
charcoal rot, wilt and yellow mold. These diseases are known to render
Groundnut production highly unstable (Mishra and Ghewande, 1989).
Approximately 5-15 % loss in the initial crop stand is due to seed-rot and
seedling collapse (Pande and Narayana Rao, 2000). Out of these diseases,
yellow mold caused by Aspergillus flavus Link ex Fr. produces aflatoxin
which is carcinogenic to both human and animals. It is estimated that about
4% of the worlds grains are lost due to biodeterioration caused by seed
borne fungi (Clerke, 1966).

Antifungal activity of extracts of weed biomass against Aspergillus flavus Link ex Fr.
In recent years much attention has been given to non were then aseptically transferred to the moist
chemical systems for seed treatment to protect them chamber plate. The plate was then incubated at room
against seed-borne pathogens. Plant extracts have temperature for 7 day and also watered regularly with
played significant role in the inhibition of seed-borne sterile distilled water. After 7 days incubation, fungus
pathogens and in the improvement of seed quality and produced large green conidial heads with spores
field emergence of plant seeds (Patel et al., 2007). which was removed aseptically, transferred to PDA
plates and slants. The fungus was identified by
In India many unwanted plants so called weeds (Plant observing colony morphology and microscopic
biomass) are very common, dominant and wide spread characteristic.
in the crop fields. In India diversity of unwanted plant
in crop fields is very common, dominant and easily The identification was confirmed with the help of
available. Weeds also occupy almost all open spaces. latest manuals, Subramanian, (1971), Neergaard and
They spread like wildfire and grow abundantly in the Mathur, (1980), Jha, (1993) and Mukadam, (1997) and
crop fields, forest and roadsides. with the help of Information Bulletin of International
Crops Research Institute for the Semi-Arid tropics
During botanical excursions to different parts of this (ICRISAT), Pantancheru, Andhra Pradesh (India). Pure
phytogeographic region every year, it is observed fact culture of the identified fungi was prepared and
that weeds produce a huge biomass. This fact maintained on PDA (Potato Dextrose Agar) slants.
promoted to work on the present research topic.
IV) Preparation of weed extracts
The present investigation was undertaken to study the Fresh samples were washed in tap water and finally
efficacy of common, dominant and easily available washed thrice using sterilized distilled water. They
weeds in the crop fields against the yellow mold were crushed in a sterilized pestle and mortar by
causing fungi, Aspergillus flavus Link ex Fr. in a cost- adding a little quantity of sterile distilled water just
effective and eco-friendly manner. enough to crush the sample easily. The extracts were
collected by filtering through the two layers of muslin
cloth. Finally, filtrates thus obtained from the leaves
MATERIAL AND METHODS were used as stock solution (Kuntal Das et al., 2010).
I) Collection of seed samples V) Studies on antifungal activity of extracts of weed

The method described by Neergaard (1977) has been biomass on fungal growth (in vitro)
adopted for the collection of seed samples. To study the antifungal mechanism of plant extract,
Accordingly, three random samples of seeds (half Kg poisoned food technique was followed as suggested by
each) were collected from oil mills, market place and Nene and Thapliyal (1982).
Oil Seed Research Station (ORS), Latur. Groundnut
cultivars used in the present study are TAG-24 and SB- Fresh weed extract stock solution were used at three
XI. concentrations (1 %, 5 % and 10 %) prepared by
mixing aseptically 1 ml, 5 ml and 10 ml of stock
II) Detection of seed mycoflora solution in 100 ml semisolid sterilized potato dextrose
The seed mycoflora was detected by using standard agar medium (Tiwari et al., 2005) at a temperature of
moist blotter paper method and agar plate methods as 30C - 40C. Three replications were maintained for
recommended by International Seed Testing each treatment. Suitable control plates were
Association (ISTA, 2003), Neergaard, (1977) and maintained. Each plate was seeded with 5 mm mycelial
Agarwal, (1981). discs aseptically taken from the periphery of 7 days
old culture and incubated at 27 C till the growth of the
III) Isolation and identification of test fungi colony touched the periphery in control plate. Mean
Infected groundnut seeds were selected for isolation of colony diameter in each case was recorded (Dange,
A. flavus by using the procedure published by ISTA 2006). The efficacy of weed extracts was expressed as
(2003). Infected seeds were first disinfected with 0.1 per cent inhibition of mycelial growth over control
% HgCl2 for 2 to 3 minutes and then washed with which was calculated by using the formula as given by
sterile distilled water 2 to 3 times. Disinfected seeds (Verma and Kharwar, 2006).

Deshmukh RS, 2017
RESULTS AND DISCUSSION TAG-24 and SB-XI were plated on moist blotters and
the percent incidence of seed-borne fungi was
I) Detection of seed mycoflora recorded and presented in table-1. From the results it
In order to study varietal variations in the seed is clear that, there was variation in the degree of
mycoflora, seeds of different varieties of Groundnut i.e. incidence of mycoflora in groundnut varieties.
Table- 1: Seed Mycoflora of Groundnut varieties TAG-24 and SB-XI (Moist Blotter Plate Method).
Sr. Seed borne Fungi Varieties of Groundnut
No. Per cent incidence of seed-borne fungi
TAG-24 SB-XI
1 Alternaria alternata (Fr.) Keissler 10 -
2 Aspergillus candidus Link 10 05
3 Aspergillus flavus Link ex Fr. 40 35
4 Aspergillus fumigatus Fresenius 10 -
5 Aspergillus niger van Tieghem 40 25
6 Aspergillus terreus Thom. 10 -
7 Fusarium oxysporum Schlechtend emend Sny. & Hans. 10 05
8 Macrophomina phaseolina (Tassi) Goldanich 10 -
9 Rhizopus nigricans Ehrenb. 20 20
10 Sclerotium rolfsii Sacc. 05 -
Table- 2: Antifungal activity of extracts of weed biomass against Aspergillus flavus Link ex Fr. by poisoned food
technique.
Mean colony diameter (in mm)
Sr.
Name of Weeds Part Used Aqueous Extracts Ethanolic Extracts
No.
1% 5% 10% 1% 5% 10%
Leaves 39 38 35 32 32 30
1 Argemone maxicana L.
Stem 38 38 38 35 32 30
Leaves 11 09 07 11 07 05
2 Commelina benghalensis L.
Stem 25 24 20 26 25 25
Leaves 37 35 35 36 32 31
3 Cynodon dactylon (L.) Pers.
Stem 39 36 35 38 35 35
Leaves 35 34 32 34 32 30
4 Cyperus rotundus L.
Rhizome 39 36 35 38 35 35
Leaves 40 39 39 41 40 40
5 Euphorbia hirta L.
Stem 42 42 40 40 40 40
Leaves 13 11 09 12 10 09
6 Parthenium hysterophorus L.
Stem 08 07 05 08 06 06
Phyllanthus amerus Leaves 41 41 41 40 36 35
7
Schumach.& Thonn Stem 42 40 40 41 40 40
Leaves 28 28 27 25 22 22
8 Portulaca oleracea L.
Stem 28 25 25 24 20 20
Leaves 37 35 35 35 32 30
9 Solanum nigrum auct.
Stem 39 38 35 35 30 30
Leaves 42 41 40 40 40 39
10 Tridex procumbens L.
Stem 45 43 42 45 40 40
11 Control 100 100
S.E. + 0.97 1.31 0.91 1.08 1.07 1.38
Leaves
C.D. at 0.05% 2.86 3.86 2.68 3.20 3.16 4.07
S.E. + 1.20 1.70 1.81 1.20 2.15 1.27
Stem
C.D. at 0.05% 3.55 5.00 5.33 3.55 6.33 3.74

Antifungal activity of extracts of weed biomass against Aspergillus flavus Link ex Fr.
Table-3: Efficacy of extracts of weed biomass against mycelial growth of Aspergillus flavus Link ex Fr. by poisoned
food technique.
Sr. Name of Weeds Part Used Per cent mycelial inhibition
No. Aqueous Extracts Ethanolic Extracts
1% 5% 10% 1% 5% 10%
1 Argemone maxicana L. Leaves 61 62 65 68 68 70
Stem 62 62 62 65 68 70
2 Commelina benghalensis L. Leaves 89 91 93 89 91 95
Stem 75 76 80 74 75 75
3 Cynodon dactylon (L.) Pers. Leaves 63 65 65 66 68 69
Stem 61 64 65 62 65 65
4 Cyperus rotundus L. Leaves 65 66 68 66 68 70
Rhizome 61 64 65 62 65 65
5 Euphorbia hirta L. Leaves 60 61 61 59 60 60
Stem 58 58 60 60 60 60
6 Parthenium hysterophorus L. Leaves 87 89 91 88 90 91
Stem 92 93 95 92 94 94
7 Phyllanthus amerus Leaves 59 59 59 60 64 65
Schumach.& Thonn Stem 58 60 60 59 60 60
8 Portulaca oleracea L. Leaves 72 72 73 75 78 78
Stem 72 75 75 76 80 80
9 Solanum nigrum auct. Leaves 63 65 65 65 68 70
Stem 61 62 65 65 70 70
10 Tridex procumbens L. Leaves 58 59 60 60 60 61
Stem 55 57 58 55 60 60
11 Control 00 00
Leaves S.E. + 1.93 2.1 2.49 2.54 2.04 2.30
C.D. at 0.05% 5.68 6.20 7.33 7.47 6.00 6.77
Stem S.E. + 2.40 1.86 2.25 2.11 2.30 2.28
C.D. at 0.05% 7.08 5.48 6.64 6.22 6.77 6.71
Groundnut variety TAG-24 showed maximum seed III) Antifungal activity of extracts of weed biomass
mycoflora including ten fungi while variety SB-XI against Aspergillus flavus Link ex Fr. by poisoned
showed presence of five fungi. The seeds of both the food technique.
varieties, TAG-24 and SB-XI dominantly showed During the present investigation it was noticed that
presence of Aspergillus flavus. the yellow mold causing fungi, Aspergillus flavus was
dominant causing heavy damage to Groundnut seeds.
II) Identification of yellow mold causing fungi Hence antifungal activity of extracts of weed biomass
Microscopic characteristics like conidial head, conidia was tested against Aspergillus flavus by Poisoned food
shape, roughness and vesicle serration, etc were taken technique.
into consideration for identification. Aspergillus flavus
Link ex Fr.:-The mycelium is found to be submerged in From the results presented in table- 2 it is clear that
the seed coat and forms a white to grey, tough mass. maximum antifungal activity was observed in 10 %
Conidiophores erect, simple, unbranched, hyaline, Aqueous and 10 % Ethanolic stem extracts of
transparent and smooth. The apex of the Parthenium hysterophorus L. (05 mm and 06 mm
conidiophores was inflated into a vesicle upon which respectively) followed by 10 % Ethanolic leaf extracts
radiating phialides are formed. Conidial heads were of Commelina benghalensis L. (05 mm) as compared
biseriate, globose to radiate often columnar, very light with control (100 mm).
to deep yellow green, olive brown often brown.
Conidia were found to be hyaline, single celled and Other test weeds do not show much antifungal activity
produced in chains. They were globose to subglobose, as compared with Parthenium hysterophorus L. and
often elliptical to pyriform and conspicuously Commelina benghalensis L.
echinulate.

Deshmukh RS, 2017
IV) Efficacy of extracts of weed biomass against University of Agri. and Tech. Pantnagar. Agric. Biology,;
mycelial growth of Aspergillus flavus Link ex Fr by 2(1&2):144-146.
poisoned food technique. Clerke JH (1966) Fungi in stored products. PANS. 13:473-
481.
The efficacy of weed extracts was expressed as per
Dange Vinod. Studies on root rot of chilli casused by
cent inhibition of mycelial growth over control which Sclerotium rolfsii Sacc. M.Sc. Thesis, University of
was calculated by using the formula as given by Verma Agricultural Sciences, Dharwad 2006.
and Kharwar (2006). From the results presented in Elwakil MA and El-Metwally MA (2001) Seed-borne fungi of
table 3, it was observed that maximum per cent peanut in Egypt: Pathogenicity and transmission. Pak. J.
Biol. Sci. 4: 63-68.
mycelial growth inhibition of Aspergillus flavus was
Embaby EM, Mona M and Abdel-Galil (2006) Seed borne
done by 10% Aqueous stem extract of Parthenium
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and 10 % Ethanolic stem extracts of Parthenium (2007) Screening of Groundnut Genotypes for Leaf
hysterophorus L. (94 %). Spots and Rust Resistance. Int. J. Sustain. Crop Prod.
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CONCLUSION properties of rhizome extracts of some medicinal plants.
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the degree of varietal resistance against the incidence V.P. Agnihotori), CBS Pub. Delhi 1989.
of seed borne fungi. Similar type of variations in Mukadam DS. The illustrated kingdom of fungi (some
selected genera). Published by Akshar Ganga prakashan,
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Aurangabad, India 1997.
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studied the seed mycoflora of Groundnut and reported Mysore, India 1980.
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control. Second Edn , Oxford and IBH Publishing Co.,
New Delhi 507; 1982.
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that infect crops. Newsletter, 20: 42-44.
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important crops. Research Bulletin 108, G.B. Pant

RESEARCH ARTICLE
Incidence of mycotoxins in poultry feeds and feed ingredients

used in Warangal (TS), India
Parvathi D, Rajender Reddy A and Krishna Reddy V
Toxicology Laboratory, Department of Botany, Kakatiya University, Warangal-506 009 (TS), India
Email Id: vkreddyku@gmail.com

Received: 12.07.2017 Incidence of mycotoxins is a major problem encountered by poultry
Accepted: 13.08.2017 industry as in most cases low grade feed ingredients are used in the
Published : 23.09.2017 formulation of feeds. Investigations carried out to assess the incidence of
mycotoxins in poultry feeds revealed that aflatoxin followed by ochratoxin
Editor:
A were most common and contaminated large number of poultry feeds and
Dr. Arvind Chavhan
feed ingredients. Most of the samples were found to be contaminated with
Cite this article as: more than one mycotoxin. Farm mixed feed samples were recorded with
Parvathi D, Rajender Reddy A and highest frequencies of mycotoxin contamination than the commercial
Krishna Reddy V (2017) Incidence of brands,
mycotoxins in poultry feeds and feed
ingredients used in Warangal (TS), Keywords: Poultry feeds, mycotoxins, aflatoxin, ochratoxin A, broilers,
India; International J. of Life Sciences, layers.
5 (3): 399-404.
Acknowledgements
Authors are thankful to the Head, INTRODUCTION
Department of Botany for providing
facilities and the financial assistance Contamination of food and agricultural commodities by various types of
of UGC is gratefully acknowledged. toxigenic molds is a serious nonetheless a widely neglected problem.
Regardless of decades of extensive research, mold infestation still remains a
challenging problem (Munkvold, 2003). It has been estimated by Food and
terms of the Creative Commons Agriculture Organization (FAO) that worldwide, approximately 25% of the
Attribution-Non-Commercial - No crops get contaminated by moulds and are affected by mycotoxins (Richard
Derivs License, which permits use et al., 1989; Rice and Ross, 1994), and the estimated loss extends to billions
and distribution in any medium, of dollars (Trail et al., 1995). Poor harvesting practices, improper drying,
provided the original work is handling, packaging, storage, and transport conditions contribute to fungal
properly cited, the use is non- growth and increase the risk of mycotoxin production. Contamination of
commercial and no modifications or agricultural commodities by fungi results not only in the downgrading
nutrient content but also cause a health hazard for human, livestock and
poultry birds. The most notable importance of contamination of agricultural
commodities by toxigenic fungi is buildup of mycotoxin concentrations to
injurious levels in food and feeds (Perrone et al., 2007). Poultry feed are
formulated from a mixture of ingredients that include cereal grains, cereal
byproducts, fats, plant protein sources, animal byproducts, vitamins,
mineral supplements, crystalline amino acids and feed additives. The
composition of the feed varies according to type of poultry bird (broiler or
layer) and also age of the bird.

Parvathi et al., 2017
Since poultry feeds are rich in all types of nutrients 1975); ochrotoxin-A (Gimeno, 1979); sterigmato-
easily to prone to microbial growth at appropriate cystine (Adey and Mateles, 1964); penicillic acid
conditions. Defects in formulations, faulty storage (Gorst-Allman and Steyn, 1979); deoxynivalenol and
conditions and unseasonal rains promote the nivalenol (Ramakrishna and Bhat, 1987);
infestation by microorganisms. Of the potential moniliformin, T-2 toxin and zearalenone (Kamimura et
contaminants, mycotoxigenic fungi are the most al., 1981)
widespread and notorious particularly in hot, humid
conditions. In view of the importance of mycotoxins in
the health of poultry birds and a range of poultry feeds RESULTS AND DISCUSSIONS
used, it was thought worthwhile to investigate the
natural incidence of mycotoxins in different poultry 1. Broiler feeds
feeds used in Warangal District, which is a major The mould infested broiler feed samples collected
center for poultry industry in Telangana State. from different broiler poultry farms of Warangal
district were analyzed for the presence of mycotoxins
and the results are presented in Table 1. From table it
MATERIALS & METHODS is evident that the contamination of different broiler
feeds with mycotoxins is a common feature. Out of 35
Isolation and enumeration of mycotoxigenic fungi broiler concentrate feed samples tested for the
associated with poultry feed, feed ingredients presence of mycotoxins, 7 samples were positive for
aflatoxins, 3 samples for ochratoxin A, 1 sample for
Study region zearlenone and 7 samples for T- 2 toxin.
Samples of commercial poultry feeds, feed ingredients Sterigmatocystin was not found in broiler concentrate
were collected from poultry farms located in Warangal feed samples. Similarly, 28 samples of broiler
district and the respective feed manufacturers located concentrate feed with maize were positive for
in other places also. In case of farm mixed poultry aflatoxins and 13 for ochratoxin A, 2 for zearalenone
feeds, individual feed ingredients along with mixed and 8 for T-2 toxin, out of 35 samples were screened.
feeds were also collected. Sterigmatocystin was not detected in broiler
concentrate feed with maize.
Sampling criteria
Samples of commercially prepared poultry feeds, farm Out of 35 broiler pellet feed samples screened for
mixed poultry feeds and feed Ingredients were presence of different mycotoxins, 9 samples were
collected through systematic random sampling from positive for aflatoxins, 2 for ochratoxin A and1 for T-2
grain markets, feed ingredient suppliers, poultry farms toxin. Zearalenone and sterigmatocystin were not
and feed manufacturers. For each sample, three initial found in the broiler pellet feed samples tested.
samples of three kg each were collected from different Similarly, out of 35 broiler pre starter crumbs samples
places. After thorough mixing, a composite sample of screened for mycotoxin analysis, 3, 2, 1 and 1 samples
1 kg placed in a sterile polythene bag was brought to were positive for aflatoxins, ochratoxin A, zearalenone
the laboratory and they were refrigerated until and for sterigmatocystin respectively. However T-2
further use. Conditions of the sample at the collection toxin was not detected in the samples screened. No
time were recorded. toxin was reported in broiler starter feed samples.
Natural incidence of mycotoxins in poultry feeds. 2. Layer feeds

Natural contamination of commercial poultry feeds Different contaminated layer feed samples were
was carried out by rapid TLC method for multi- collected from different layers poultry farms of
mycotoxin analysis as suggested by Devegowda and Warangal district and screened for presence of
Raju (2005). different mycotoxins and the results are presented in
Table 2. From the table it is evident thatout of the
Different mycotoxins produced by species of different layer feed samples screened for presence of
Aspergillus, Fusarium and Penicillium were detected by mycotoxin contamination, homemade feed (mixed at
standard methods. Aflatoxins (Stack and Pohland, the farm) samples were highly contaminated with

Incidence of mycotoxins in poultry feeds and feed ingredients used in Warangal (TS), India
different mycotoxins. Screening of 35 mixed layer feed samples indicated that 25 samples were positive for
Table 1 : Natural contamination of different broiler feeds with mycotoxins
Name of the broiler feed Number of feed Number of positive % of incidence Toxin detected
sample tested samples tested feed samples
Broiler concentrate feed 35 7 20.0 Aflatoxins
3 8.6 Ochratoxin A
1 2.9 Zearalenone
7 20.0 T-2 toxin
ND -- Sterigmatocystin
Broiler concentrated 35 28 80.0 Aflatoxins
feed with maize 13 37.1 Ochratoxin A
2 5.7 Zearalenone
8 22.9 T-2 toxin
Broiler pellet feed 35 9 25.7 Aflatoxins
2 5.7 Ochratoxin A
ND -- Zearalenone
1 2.9 T-2 toxin
Broiler pre starter
crumbs 35 3 8.6 Aflatoxins
2 5.7 Ochratoxin A
1 2.9 Zearalenone
ND -- T-2 toxin
1 2.9 Sterigmatocystin
ND = Not detected
Table 2 : Natural contamination of different layer feeds with mycotoxins

Name of the layer Number of feed Number of % of Toxin detected
feed sample tested smaples tested positive feed samples incidence
Home made feed 35 25 71.4 Aflatoxins
(mixed feed) 13 37.1 Ochratoxin A
2 5.7 Zearalenone
5 14.3 T-2 toxin
Chick feed 35 5 14.3 Aflatoxins
2 5.7 Ochratoxin A
ND -- Zearalenone
1 2.9 T-2 toxin
Developer feed 35 8 22.9 Aflatoxins
3 8.6 Ochratoxin A
1 2.9 Zearalenone
2 5.7 T-2 toxin
Grower feed 35 11 31.4 Aflatoxins
3 8.6 Ochratoxin A
1 2.9 Zearalenone
-- -- T-2 toxin
Perlay feed 35 9 25.7 Aflatoxins
ND -- Ochratoxin A
ND -- Zearalenone
3 8.6 T-2 toxin

Table 3 : Natural contamination of different poultry feed ingredients with mycotoxins

Name of the Nummber of Number of % of incidence Toxin detected
feed ingredient tested samples tested positive samples
Maize seed 50 34 68.0 Aflatoxins
13 26.0 Ochratoxin A
2 4.0 Zearalenone
8 16.0 T-2 toxin
Sunflower meal 50 22 44.0 Aflatoxins
3 6.0 Ochratoxin A
1 2.0 Zearalenone
4 8.0 T-2 toxin
Groundnut cake 50 28 56.0 Aflatoxins
13 26.0 Ochratoxin A
3 6.0 Zearalenone
2 4.0 T-2 toxin
Deoiled rice bran 50 14 28.0 Aflatoxins
8 16.0 Ochratoxin A
6 12.0 Zearalenone
4 8.0 T-2 toxin
Fish meal 50 3 6.0 Aflatoxins
ND -- Ochratoxin A
ND -- Zearalenone
ND -- T-2 toxin
Soybean meal 50 9 18.0 Aflatoxins
6 12.0 Ochratoxin A
5 10.0 Zearalenone
4 8.0 T-2 toxin
aflatoxins and 13 samples for ochratoxin A, 2 for toxin. Sterigmatocystin was absent in developer feed
zearalenone, 5 for T-2 toxin and 3 for sterigmatocystin. samples.
This can be attributed for the usage of damaged feed Screening of 35 grower feed samples revealed that 11
ingredients by the layer farmers to reduce the feed samples were positive for aflatoxins, similarly 3 for
cost. ochratoxin A, 1 for zearalenone and 1 for
sterigmatocystin. T-2 toxin was not detected in grower
When 35 chick feed samples were screened for their feed samples screened. Out of of 35 prelay feed
natural contamination of different mycotoxins 5, 2 and samples tested 9, 3 samples were positive for
1 sample were positive for aflatoxins, ochratoxin A and aflatoxins, T-2 toxin respectively. Interestingly,
T-2 toxin respectively. Zearalenone and ochratoxin A, zearalenone and sterigmatocystin were
sterigmatocystin were not traced in the chick feed not detected in the samples screened.
samples screened. Similarly, 35 developer feed
samples screened for presence of mycotoxin, 3.Feed ingredients
contamination 8 samples were positive for aflatoxins, Feed ingredients are the key resources for
3 for ochratoxin A, 1 for zearalenone and 2 for T-2 contamination of feeds with mycotoxins. Hence, it was
felt necessary to screen the different feed ingredients

Incidence of mycotoxins in poultry feeds and feed ingredients used in Warangal (TS), India
used in Warangal district, and the results are shown in home-brew methods for detection of mycotoxins in the
the Table 3. From the table it is evident that most of feeds prepared at farm level.
the feed ingredients screened were positive for
different mycotoxins. Out of 50 samples of maize
screened, 34 samples are positive for aflatoxins and 13 Conflicts of interest: The authors stated that no conflicts of
for ochratoxin A, 2 for zearalenone and 8 for T-2 toxin, interest.
and 9 for sterigmatocystin.
Out of 5 samples of sunflower meal screened for REFERENCES

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Adye J and Mateles RC (1964) Incorporation of labeled
aflatoxins, and 3 for ochratoxin A, 1 for zearalenone
compounds into aflatoxins. Biochimica et Biophysica
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positive for sterigmatocystin. Similarly,out of 50 Dalcero A, Magnoli C, Hallak C, Chiacchiera S, Palacio G and
groundnut cake samples screened for presence of Rosa CDR (2002) Detection of ochratoxin A in animal
mycotoxins,28,13,3,2 samples were positive for feeds and capacity to produce this mycotoxin by
Aspergillus section Nigri in Argentina. Food Additives
aflatoxins, ochratoxin A, zearalenone, T-2 toxin and Contaminants 19, 1065-1072.
respectively. Sterigmatocystin was not recorded in the Devegowda G and Raju MVLN (2005) Manual on mycotoxin
samples screened. quantification in foods and feeds. Veterinary College,
University of Agricultural Sciences, Karnataka
Screening of 50 deoiled rice bran samples for their University, India.
natural contamination of mycotoxins indicated that 14 Gimeno A (1979) Thin layer chromatographic determination
of aflatoxins, ochratoxins, sterigmatocystin,
samples were positive for aflatoxins and 8 for zearalenone, citrinin, T-2 toxin, diacetoxyscirpenol,
ochratoxin A, 6 for zearalenone, 4 for T-2 toxin and 1 penicillic acid, patulin and penitrem A. Journal-
for sterigmatocystin. Similarly, in 50 fish meal samples Association of Official Analytical Chemists 62, 579-585.
tested only 1 sample was positive for sterigmatocystin, Gorst-Allman CP and Steyn PS (1979) Screening methods for
the detection of thirteen common mycotoxins. Journal of
3 samples were positive for aflatoxins, ochratoxin A,
Chromatography A 175, 325-331.
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Kamimura H, Nishijima M, Yasuda K, Saito K, Ibe A,
fishmeal sample. Similarly, when 50 soybean meal Nagayama T, Oshiyama H and Naoi Y(1981)
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Incidence of fumonisins, moniliformin and Fusarium
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Munkvold GP(2003) Cultural and genetic approaches to
Studies on natural incidence of mycotoxins in poultry managing mycotoxins in maize. Annual Review of
Phytopathology 41, 99-116.
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Oliveira GR, Ribeiro JM, Fraga ME, Cavaglieri LR, Direito GM,
are most common and contaminated large number of
Keller K., Dalcero AM and Rosa CA (2006) Mycobiota in
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samples were contaminated with more than one fumonisins and zearalenone in the Rio de Janeiro State,
mycotoxin. Farm mixed feed samples were recorded Brazil. Mycopathologia 162, 355-362.
with highest frequencies of mycotoxin contamination Perrone G, Susca A, Cozzi G, Ehrlich K, Varga J, Frisvad JC,
Meijer M, Noonim P, Mahakarnchanakul W and Samson
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Morocco. Food Control 17, 868-874.

RESEARCH ARTICLE
Phytochemical Screening and Antibacterial Activity of Bryophytes
Kadam Pratima S
Department of Botany, Prof. Ramkrishna More ACS College, Akurdi, Pune 411044, India
Email: drpskadam@gmail.com

Received: 22.07.2017 In this paper we study antibacterial activity and phytochemical screening of
Accepted: 29.08.2017 the bryophytes. The bryophytes possess some bioactive compounds and
Published : 23.09.2017 therefore are medicinally very useful plants. They contain pools of
secondary metabolites, which are unique .The antimicrobial activity was
Editor:
assessed using agar well diffusion method against the two bacterial strains
Dr. Arvind Chavhan
from the bryophytes viz Anthoceros erectus, Asterella angusta, Cyathodium
Cite this article as: tuberosum, Plagiochasma articulata and Targionia hyphophylla the studies
Kadam, Pratima S (2017) of phytochemical constituents revealed presence of alkaloids, flavonoids,
Phytochemical Screening and coumarins, phenols, tannins, steroids, and sugars. The antimicrobial activity
Antibacterial Activity of Bryophytes is due to the presence of secondary metabolites. It was highest in Targionia
International J. of Life Sciences, 5 (3): hyphophylla and least in Cyathodium tuberosum. The above observations
405-408. suggest that due to significant antibacterial activity of the bryophytes they
have good potential in drug development.
Acknowledgement:
The author is thankful to BCUD, Keywords: Antimicrobial activity, Phytochemical screening, Bryophytes
SPPU, Pune, for funding this
research work.
Copyright: 2017| Author (s), This INTRODUCTION

Bryophytes are the oldest land plants, which include liverworts, hornworts
and mosses. This group of vascular plants include about 25,000 to 28,000
species and they grow in shady locations. They are herbaceous and absorb
provided the original work is water and mineral nutrients mainly through leaves. They have an
properly cited, the use is non- interesting feature, which is about not getting attacked by bacteria, fungi or
commercial and no modifications or pests. They are potential source of medicine as they contain secondary
adaptations are made. metabolites. The antibiotic properties are due to the biologically active
compounds. Bryophytes are a rich reservoir of biological active compounds
such as terpenoids, flavonoids, alkaloids, glycosides, saponins,
anthroquinons, sterols and other aromatic compounds. They also possess
anticancer and antimicrobial activity due to their unique chemical
constituents. In the present investigation the bryophytes were screened for
their antibacterial activities and phytochemical constituents. In this paper
we study antibacterial activity and phytochemical screening of the
bryophytes. This study would enable investigation of the use of bryophytes
in drug development.

Kadam Pratima S, 2017
MATERIAL AND METHODS methanol and ethanol (Hi-Media), was used as ve

control were added in the wells. All the plates were
The bryophytes viz Anthoceros erectus, Asterella kept at low temperature for 1-1 hrs for sample
angusta, Cyathodium tuberosum, Plagiochasma diffusion and incubated at 35C for 48 hrs. After
articulate and Targionia hyphophylla, were collected incubation, the zone of inhibition was measured.
from Lonavla.
Statistical analysis: Statistical analysis was done
Plant extraction: The plant material was carefully using one-way analysis of variance (ANOVA). The
picked from the soil and washed thoroughly with quantity P < 0.05 was considered significant.
distilled water to remove the adhering soil or
extraneous dust particles. A quantity of 50 gm air Phytochemical screening: The aqueous extracts of all
dried plant material along with rhizoids was powered the selected bryophytes were subjected to various
in mortar with pestle extracted in methanol and phytochemical tests to detect the presence of certain
ethanol. These extracts were used for further studies. bioactive compounds (Trease, 2002).
Test microorganisms: The bacterial strains were

procured from NCIM, NCL, Pune. In vitro antibacterial RESULTS
activity was tested against Staphylococcus aureus and
Bacillus subtilis. The antibacterial activity has been depicted in Fig. 1.
The results reveal that the bryophytes Asterella
Determination of antimicrobial activity: Plant angusta had greater antibacterial activity in methanol
extracts were tested for antibacterial activity through extract and in Targionia hyphophylla and Plagiochasma
agar-plate diffusion method using 100 l of suspension articulata the antibacterial activity was greater in
of the test microorganisms. Bacterial strains were ethanol extract. In Anthoceros erectus the antibacterial
grown on nutrient agar plates. Sterile nutrient agar activity of both methanol and ethanol was less and
plates were prepared, and 48 hours cultured least in Cyathodium tuberosum The phytochemical
suspensions were made and inoculated on sterile agar screening is depicted in Table 1. It indicated the
medium in the respective culture plates. The 10 mm presence of secondary metabolites like alkaloids,
sterile cork borer was used for making wells. The phenols, tannins, flavonoids, coumarins, steroids, and
extracts, 0.1 and 0.2 ml were added in assay well sugars; whereas carboxylic acid, resins, saponins were
carefully. A quantity of 50 g ml1 tetracycline and not present.
ampicillin (Hi-Media), which served as +ve control
Table 1: Phytochemical analysis of selected bryophytes
Phytochemical Anthoceros Asterella Cyathodium Plagiochasma Targionia

constituents erectus angusta tuberosum articulata hyphophylla
Alkaloids + + - - +
Carboxylic acids - - - - -
Coumarins + + - + +
Flavonoids + + - + +
Phenols + + + + +
Resins - - - - -
Saponins - - - - -
Steroids + - - + +
Tanins - - - + +
Sugars + + + + +

Phytochemical Screening and Antibacterial Activity of Bryophytes
Fig. 1: Antibacterial activity of methanol and ethanol extracts of some bryophytes
DISCUSSION appendiculatum, Conocephalum coricum, Mnium

marginatum, and Byrum argenteum have antibacterial
The present study reports that there is varying level of activity and their findings support the use of
activity in the test species. This suggests that the bryophytes in traditional medicine for treating burn
extracts in both the solvents have a broad spectrum infections.
activity, hence there is a need to investigate further for
their use as for antibacterial agents, which could help The results of the phytochemical screening reveals
in new drug development (Sawant et al., 2010). The that the phytochemical compounds may be
antimicrobial activity carried out by Oyesiku and Caleb responsible for antimicrobial activity of the
(2015) in three mosses also reveal that ethanol bryophytes and they are alkaloids, flavonoids,
extract was found to be more active than the other two quinones, resins, steroids, tanins and sugars. Batish et
and are of the opinion that these extracts show al. (1997), Ahmed et al. (1998), and Kumaraswamy
antibiotic activity. This suggests that specific and Satish (2008) are of the opinion that the activity
antibacterial compounds isolated by ethanol are more may be due to presence of various secondary
effective against specific bacteria. Deora and Rathore metabolites. Deora (2015) has observed that selected
(2013) have revealed that Plagiochasma articulata bryophytes showed the presence of terepenoids,
crude extract adversely affected the bacterial growth. flavonoids, steroids, and glycosides were present
whereas, whereas alkaloids, saponins and
Bodade et al. (2008) also have observed that the anthroquinons were not present and have further
ethanolic extract was more active than other stated that these chemical compounds could be potent
fractioned extracts and Plagiochasma articulata was antimicrobial agents to treat plant diseases.
most active against bacteria and fungi. They are of the
opinion that antimicrobial activity may be due to
secondary metabolites. Similar observations have been CONCLUSION
made by Nikolajeva et al. (2012) in different
bryophytes. They have found that 73% ethanolic In this research work the antimicrobial activity in
extracts exhibited antibacterial activity and it was also bryophytes was assessed using agar well diffusion
higher as compared with aqueous extracts. Studies method against the two bacterial strains. The
carried out by Singh et al. (2011) also reveal that all bryophytes considered were Anthoceros erectus,
the four different bryophyte, namely, Plagiochasma Asterella angusta, Cyathodium tuberosum,

Kadam Pratima S, 2017
Plagiochasma articulata and Targionia hyphophylla. Deora GS and Rathore MS (2013) Antimicrobial Activity of
The studies of phytochemical constituents revealed Certain Bryophytes, Biosciences Biotechnology
Research Asia, 10(2), 705-710.
presence of alkaloids, flavonoids, coumarins, phenols,
tannins, steroids, and sugars in the selected Deora GS (2015) Phytochemical Screening and Antibacterial
Studies of Certain Bryophytes against Some
bryophytes. The antimicrobial activity is due to the Phytopathogenic Bacteria, Int. J. Pharm. Sci. Rev. Res.,
presence of secondary metabolites. It was observed to 35(1), 74-77.
be highest in Targionia hyphophylla and least in Kumaraswamy MV and Satish S (2008) Antioxidant and
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RESEARCH ARTICLE
Diversity, Distribution and Status of the Amphibian fauna of Sangli

district, Maharashtra, India
Sajjan MB1*, Jadhav BV2 and Patil RN1
1Department of Zoology, Sadguru Gadage Maharaj College, Karad - 415124, (M.S.), India
2Department of Zoology, Balasaheb Desai College, Patan - 415206, (M.S.), India
*Corresponding author E-mail: sajjan_mb73@yahoo.com

Received: 26.07.2017 30 species of amphibians were reported during a survey belonging to 19
Accepted: 20.08.2017 genera of 9 families and 2 orders from Sangli district, Maharashtra, India,
Published : 23.09.2017 during June 2013 to May 2017. Out of 30 species recorded, 19 species are
endemic to Western Ghats. All of the tehsils in this district except Shirala fall
Editor:
under semi arid zone having rich amphibian diversity. Shirala tehsil is
Dr. Arvind Chavhan
flanked by Western Ghats with high rainfall and humidity harbouring
Cite this article as: highest number of species, while Atpadi tehsils is a drought prone zone
Sajjan MB, Jadhav BV and Patil RN with the lowest number of species. The highest numbers of species are
(2017) Diversity, Distribution and reported at 1100m asl and the lowest number of species in the area below
Status of the Amphibian fauna of 600m asl. Along with checklist, information about the habitat, rainfall,
Sangli district, Maharashtra, India, temperature, distribution and status of amphibians in the district are given.
409-419. Keywords: Amphibian diversity, distribution, status, habitats, Sangli
district

Attribution-Non-Commercial - No INTRODUCTION
and distribution in any medium, The diversified topography, geographic location, high rainfall and humidity
have likely to be supported to the survival of numerous species of
amphibians in the Western Ghats and the Eastern Himalayas (Andrews et
al., 2005). Amphibian of India comprises of about 405 species (Dinesh et al.,
2017), however the exact number of species has not been known since new
species of a amphibians are being discovered. The amphibian fauna of
British India was first published by Boulenger (1890). In India, several
taxonomic revisions and new species have been described by Biju and
Bossuyt (2009), Biju et al. (2014), Dahanukar et al. (2016), Garg and Biju
(2017), Giri et al. (2004), Kuramoto et al. (2007) and Padhye et al. (2014,
2015 and 2017).
In the Maharashtra Yazdani and Mahabal (1976) have listed the 11species
of amphibians from Pune. Ravichandran and Pillai (1990) have reported 13
species of amphibians from Maharashtra including 1 new species. Sekar
(1999) has given a list of 34 species of amphibians with four new records

Sajjan et al., 2017
from Maharashtra. Kamble (2002) enlisted 11 species (rainfall below 1000mm/year) and Shirala tehsil
of amphibians from Ujani wetland, Solapur district. flanked by the Western Ghats on the western side with
Jadhav et al. (2012) studied an amphibian fauna from the heavy rainfall zone (rainfall above 3000 mm/year).
Koyana, Patan Tehsil of northern Western Ghats,
Maharashtra. Padhye et al. (2012) represented 53 The present study has been carried out from June
species of amphibians from Maharashtra. Prasad et al. 2013 to May 2017. Field studies were mainly done by
(2013) reported 37 species of amphibians from Satara visits to all ten tehsils of Sangli district and to all the
tehsil. protected areas, during rainy and dry season both in
day and night. At all locations, intensive search for
Pande and Pathak (2005) reported 5 species of amphibians was undertaken by visual encounters
amphibians from the Chandoli National Park. Lawate method; the standard method formulated for
and Mule (2009) published a checklist of herpatofauna measuring and monitoring the amphibian diversity by
of the Chandoli National Park, which contains16 IUCN/SSC- DAPTF (Crump, Heyer et al., 1994). Here all
species of amphibians. Jadhav et al., (2009) reported possible sites such as the river bank, near water
Gegeneophis peters from Walwa, Dist-Sangli, bodies, along streams, in agricultural lands, grasslands,
Maharashtra. Kumbar and Patil (2010) have given under leaf litter, on tree trunks, on foliages, under
checklist of 9 anuran species from Palus tehsil stones, logs, rock crevices and decaying vegetation
including Sagareshwar wildlife sanctuary in the Sangli were searched. The searching is made of frogs and
district. Abdar (2014) has given a list of 4 species of toads using torch lights during night. On every
amphibians of the Chandoli National Park. More amphibian sighting, information on species, habitats,
(2015) reported 15 species of amphibians from Sangli microhabitat and altitude were recorded. All
district. However, most of the studies on amphibians amphibians observed during the study were
in this district are limited to short surveys. Hence the photographed and after taking morphometric
present survey was undertaken to make an extensive measurements they are released back into their
and systematic study of the amphibian fauna of Sangli natural habitat, as per the standard methods for
district with special reference to their diversity, amphibians (Crump, Heyer et al., 1994). Specimen
distribution, habitat and status. This survey provides identification was made on the basis of morphometry,
baseline data and scientific information for calls, available literature Boulenger (1890), Chanda
conservation of amphibians from arid zone. (2002), Daniels (2005), Dutta (1997), Gururaja (2012),
Inger & Dutta (1986), and with the help of other
taxonomists. The nomenclatures of species were
MATERIAL AND METHODS updated with the checklist by Dinesh et al. (2017) and
Frost (2017).
Sangli district, a part of the Deccan plateau lies
between latitude 16052' and 16087' N and longitude
74034' and 74056' E in the Western part of RESULTS AND DISCUSSION
Maharashtra. Sangli district is one of the largest
drought prone districts in Maharashtra state of India A total of 30 species of amphibians were recorded
with an area 8601.5 sq. Kms. It includes ten Tehsils viz. from various parts of Sangli district during the survey
Shirala, Walwa, Palus, Kadegaon, Khanapur (Vita), belonging to 19 genera of 9 families and 2 orders
Atpadi, Tasgaon, Miraj, Kavathe-Mahankal and Jath. (Table 1). Considering number of species in each
Altitude of the district is 500 - 1100m asl. and average family Bufonids with 3 species, Dicroglossids 9 species,
temperature is 300C. Climatically it falls under the rain Microhylids 4 species, Nyctibatrachids 2 species,
shadow region of Sahyadri Mountain. The monsoon is Ranids 3 species, Ranixalids 3 species, Rhacophorids 3
worth four months of rainy season followed by eight species and 3 species of Caecilians (Table 1). Of these
months of dry period of winter and summer. The 19 species recorded during the study are endemic to
rainfall decreases from West (Sahyadri ranges) to the the Western Ghats. 30 species of amphibians of Sangli
East. The Khanapur (Vita), Atpadi, Tasgaon, Miraj, district fall under the various categories of the IUCN
Kavathe-Mahankal and Jath tehsils are drought prone red list; endangered 2, critically endangered 1,
zone (rainfall below 500 mm/year), Walwa, Palus, vulnerable 3, least concerned 17, data deficient 1, near
Kadegaon tehsils have a moderate rainfall zone threatened 2 and 4 not assessed. 8 species were

Diversity, Distribution and Status of amphibian fauna
Plate 1: Amphibian fauna of Sangli District

Sajjan et al., 2017
Plate 2: Amphibian fauna of Sangli District

Table 1. Checklist of amphibian fauna of Sangli district with distribution, status, endemism, IUCN status and WPA- cites.
www.ijlsci.in
Sr. Name of Species Common Names Location of Species Status Endemism IUCN Red WPA-
No. (Tehsils) Lest Cites
ORDER: ANURA (Fischer von Waldheim)
FAMILY: BUFONIDAE (Gray)
1 Duttaphrynus melanostictus (Schneider Common Indian Toad 1, 2, 3, 4, 5, 6, 7, 8, 9, Abundant NE LC
1799) (Common Asian Toad) 10
2 Duttaphrynus stomaticus Marbled Toad 3, 4, 5, 6, 7, 8, 9, 10 Abundant NE LC
(Lutken, 1862) (Indus Valley Toad)

3 Xanthophryne koynayensis Koyna Toad 1 Rare EWG EN
(Soman, 1963)
FAMILY: DICROGLOSSIDAE (Anderson)
4 Euphlyctis cyanophlyctis Indian Skittering Frog 1, 2, 3, 4, 5, 6, 7, 8, 9, Abundant NE LC Sch. IV
(Schneider, 1799) (Indian Skipper Frog) 10 APP.II
5 Fejervarya caperata Canara Cricket Frog 1 Rare EWG Not
(Kuramoto, Joshy, Kurabayashi and Sumida, Assessed
2007)
6 Fejervarya cepfi CEPF Burrowing Frog 1 Rare EWG Not
(Garg and Biju, 2017) Assessed
7 Fejervarya keralensis Kerala Warty Frog 1, 2, 3, 4, 5, 6, 7, 8, 9, Abundant EWG LC Sch. IV
(Dubois, 1981) (Verrucose Frog) 10
8 Fejervarya syhadrensis Bombay Wart Frog 1, 2, 3, 4, 5, 6, 7, 8 Abundant NE LC
(Annandale, 1919)
9 Hoplobatrachus tigerinus Indian Bull Frog 1, 2, 3, 4, 5, 6, 7, 8, 9, Abundant NE LC Sch. IV
(Daudin, 1802) 10 App.II
10 Sphaerotheca breviceps Indian Burrowing 1, 2, 3, 4, 5, 6, 7, 8, 9, Abundant NE LC
(Schneider, 1799) Frog 10
11 Sphaerotheca dobsonii Dobsons Burrowing 1,2 Rare EWG LC
(Boulenger, 1882) Frog
12 Sphaerotheca pashchima Western Burrowing 1, 2, 3, 4, 5, 6, 7, 8, 9, Abundant NE Not
(Padhye, Dahanukar, Sulakhe, Dandekar, Frog 10 Assessed
Limaye and Jamdade, 2017)
413

414 Sajjan et al., 2017
Table 1: continued...
FAMILY: MICROHYLIDAE (Gunther)
13 Microhyla ornata Ornate Narrow- 1, 2, 3, 4, 5, 6, 7, 8, 9, Abundant NE LC
(Dumeril and Bibron, 1841) mouthed Frog 10
14 Uperodon globulosus Grey Balloon Frog 1, 2, 3, 4, 8 Common NE LC
(Gunther, 1864)
15 Uperodon mormorata Marbled Ramanella 1, 2 Rare EWG EN
(Rao, 1937)
Int. J. of Life Sciences, Vol. 5(3) September, 2017
16 Uperodon systoma Marbled Balloon Frog 3, 4, 5, 6, 7, 8, 9, 10 Abundant NE LC Sch. IV

(Schneider, 1799)
FAMILY: NYCTIBATRACHIDAE (Blommers-Schlosser)
17 Nyctibatrachus humayuni (Bhaduri and Bombay Night Frog 1 Rare EWG VU
Sajjan et al., 2017

Kripalani, 1955)
18 Nyctibatrachus petraeus Castle Rock Night 1 Rare EWG LC
(Das and Kunte, 2005) Frog
FAMILY: RANIDAE (Rafinesque)
19 Clinotarsus curtipes Bicoloured Frog 1 Rare EWG NT Sch. IV
(Jerdon, 1853)
20 Hydrophylax bahuvistara (padhye, Jadhav, Fungoid Frog 1, 2 Rare EWG LC Sch. IV
Modak, Nameer and Dahanukar, 2015)
21 Indosylvirana caesari (Biju, Garg, Mohony, Bronzed Frog 1 Rare EWG NT Sch. IV
Wijayathilaka, Senevirathne and (Maharashtra Golden
Meegaskumbura, 2014) Backed Frog)
FAMILY: RANIXALIDAE (Dubois)
22 Indirana beddomii Beddomes Leaping 1 Rare EWG LC Sch. IV
(Gunther, 1875) Frog
23 Indirana chiravasi (Padhye, Modak and Amboli Leaping Frog 1 Rare EWG Not
Dahanukar, 2014) Assessed
24 Indirana leithii Leiths Leaping Frog 1 Rare EWG VU
(Boulenger, 1888)

www.ijlsci.in Diversity, Distribution and Status of amphibian fauna
Table 1: continued
FAMILY: RHACOPHORIDAE Hoffman
25 Polypedates maculatus Common Indian Tree 1, 2, 3, 4, 8 Common NE LC
(Gray, 1834) Frog (Chunam Frog)
26 Pseudophilautus amboli Amboli Bush Frog 1 Rare EWG CE
(Biju and Bossuyt, 2009)
27 Raorchestes bombayensis Bombay Bush Frog 1, 2, 3, 4, 8 Common EWG VU

(Annandale, 1919)
ORDER: GYMNOPHIONA (Muller)

FAMILY: ICHTHYOPHIIDAE (Taylor)
28 Ichthyophis beddomei Beddomes Caecilian 1 Rare EWG LC
(Peters, 1879)
29 Ichthyophis bombayensis Bombay Caecilian 1 Rare EWG LC
(Taylor, 1960)
FAMILY: INDOTYPHLIDAE (Lescure, Renous, and Gasc)
30 Indotyphlus maharashtraensis Maharashtra Caecilian 1 Rare EWG DD
(Giri, Gower and Wilkinson, 2004)
_________________
1-Shirala, 2-Walwa, 3-Palus, 4-Kadegaon, 5-Khanapur, 6-Atpadi, 7-Tasgaon, 8-Miraj, 9-Kavathe-Mahankal , 10-Jath
Rare- species found in 1-2 tehsils, Common- species found in 3-6 tehsils and Abundant- species found in more than 6 tehsils
EWR-Endemic to Western Ghats, NE- Non Endemic
EN- Endangered, CE- Critically Endangered, VU- Vulnerable, LC- Least Concerned, DD-Data Deficient, NT- Near Threatened.
Schedules IV- Indian Wildlife (protection) Act
App. II of CITES - The Conservation on International Trade in Endangered Species of Wild Flora and Fauna.
415

Sajjan et al., 2017
Table 2. Amphibian fauna of Sangli district with habitat, microhabitat, altitude range, annual temprature and annual rainfall
416
Sr. Name of Species Habitat Microhabitat Altitude (m) Mean Annual Annual
No. Temp. (C) Rainfall (mm)
Min Max Min Max Min Max
1 D. melanostictus Terrestrial, near On bare land, under stone, in crevices, under street 500 1100 17 40 500 3000
human habitats lamps, near water bodies, agriculture land
2 D. stomaticus Terrestrial, near On bare land, under stone, near water bodies 500 1100 25 35 500 2000
human habitats agriculture land, grass land
3 X. koynayensis Terrestrial, On On plateau, Under rock, in crevices, in grasses 900 1200 18 26 1500 3000
lateritic rock surrounded by forest
4 E. cyanophlyctis Aquatic, littoral Found in almost all kinds of fresh water bodies 500 1100 17 38 500 3000
Int. J. of Life Sciences, Vol. 5(3) September, 2017
5 F. caperata Semi aquatic Wet soil, in grasses near water bodies, paddy and 600 800 20 24 1000 1500
sugar cane field, on forest floor
6 F. cepfi Semi aquatic, Wet soil, on forest floor, forest edge, in grasses near 650 1100 17 27 1500 3000
burrowing frog water bodies, paddy and sugar cane field
Sajjan et al., 2017

7 F. keralensis Semi aquatic Wet soil, in grasses near water bodies, paddy and 500 1000 17 38 500 3000
sugar cane field, in swamp area, in villages
8 F. syhadrensis Semi aquatic, near Wet soil, in grasses near water bodies, paddy and 500 1000 20 30 500 2000
human habitation sugar cane field, in swamp area, in villages
9 H. tigerinus Semi aquatic, near Agricultural fields, in ponds, lakes, wells and on forest 500 1100 17 40 500 3000
human habitation floor
10 S.breviceps Terrestrial, near Moist soil, near water bodies, in agricultural land, on 500 1000 17 40 500 2000
human habitation bare ground. Burrowing species
11 S. dobsonii Terrestrial, burrowing In leaf litter, on forest flour, moist soil close to water 500 1050 22 27 1000 3000
frog bodies, in paddy field, forest edge habitat
12 S. pashchima Terrestrial, near Moist soil, near water bodies, in agricultural land, on 500 1000 17 40 500 2000
human habitation bare ground. Burrowing species
13 M. ornata Semi aquatic, near In grasses, in and near water bodies, under stones, in 500 1050 18 38 500 2500
human habitation crevices, on bare ground
14 U. globulosus Terrestrial, Termite Mountain, On bare ground, near water 650 1000 22 30 1000 2500
burrowing frog bodies, agriculture land
15 U. mormorata Terrestrial, burrowing On wet soil, leaf litter, in paddy and sugar cane field, 660 1000 22 24 1000 3000
frog in forest

Table 2. Continued
www.ijlsci.in
Sr. Name of Species Habitat Microhabitat Altitude (m) Mean Annual Annual Rainfall
No. Temp. (C) (mm)
Min Max Min Max Min Max
16 U. systoma Terrestrial, , near Termite Mountain, On bare ground, near water 500 634 17 38 500 1000
human habitation, bodies, agriculture land
burrowing frog
17 N. humayuni Aquatic, forest Near shallow water streams, in rock crevices, in 560 1100 21 25 2000 3000
dwelling perennial stream, on leaves over hanging on running
water
18 N. petraeus Aquatic, forest In shallow water streams, in rock crevices, in 562 600 25 25 2000 3000

dwelling perennial stream, on leaves over hanging on runing
water, under stones

19 C. curtipes Semi aquatic, forest Forest floor, leaf litter, found near water bodies, slow 630 1100 17 27 1000 3000
dwelling running streams
20 H. bahuvistara Semi aquatic found near water bodies, forest floor, leaf litter, in 600 1000 20 30 1000 2000
paddy and sugar cane field
21 I. caesari Semi aquatic, Found near water bodies, forest floor, leaf litter, slow 630 1100 17 25 1000 3000
flowing streams
22 I. beddomii Terrestrial On moist rock, on forest floor, leaf litter, in grasses 550 1200 17 28 1000 3000
near water bodies
23 I. chiravasi Terrestrial On tree trunk, on moist rock, on forest floor, leaf 500 1000 24 25 1000 3000
litter, on moist soil, on wet clifts
24 I. leithii Terrestrial On tree trunk, on wet rock, on forest floor, leaf litter, 590 1000 22 26 1500 3000
in grasses, on moist soil
25 P. maculatus Arboreal On herbs, shrubs, trees, on sugarcane plantation 500 1100 20 30 500 1500
26 P. amboli Arboreal On herbs, shrubs, trees in forest 500 1000 24 25 1000 3000
27 R. bombayensis Arboreal On herbs, shrubs, trees, on sugar cane plantation and 500 1300 18 25 1000 3000
on ground
28 I. beddomei Fossorial In moist loose soil, in forest, in marshes, cultivated 1000 1200 14 27 1000 3000
land, in leaflitter
29 I. bombayensis Fossorial In humus rich soil, in leaf litter in forest, cultivated 800 1550 19 27 1500 3000
land, under rocks, in marshes
30 I. maharashtraensis Fossorial Under moist soil: in forest, under rocks, on plateau 630 1000 25 25 2000 3000
417

Sajjan et al., 2017
included in the schedules IV of Indian Wildlife Dr. Ramdas Bodare, Head, Department of the Zoology,
(protection) act and two species come under Appendix Sadguru Gadage Maharaj College, Karad, Dist- Satara
II of CITES. The highest number of amphibian species and the Principal Dr. S. D. Pawar Balasaheb Desai
was recorded from Shirala tehsil (28 species), while College, Patan, Dist- Satara for providing laboratory
the lowest number of species was observed in Atpadi facilities for this work. SMB is thankful to I/C Principal
tehsil (8 species). Status of amphibians shows that 10 Dr. Vitthal Dhekale and Mr. Karennvar M.H., Head,
species are abundant, 3 are common and 17 species Department of Zoology, Raje Ramrao Mahavidyalaya
are rare in the study area. Jath, Dist- Sangli for giving moral support.
Duttaphrynus stomaticus, Fejervarya caperata, Conflicts of interest: The authors stated that no conflicts of
Fejervarya cepfi (Garg and Biju 2017), Sphaerotheca interest.
pashchima (Padhye et al. 2017), Uperodon mormorata,
Uperodon systoma, Pseudophilautus amboli and
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new cryptic species. Current Herpetology Journal, 26(1):
81-105.
Lawate DV and Mule MB (2009) Herpatofauna of
Chandoli National Park of Western Ghats of
Maharashtra. Journal of Environmental Science, vol.2:
33-36.
More S (2015) Amphibian diversity from Sangli District
(MS, India). World Journal of Pharmaceutical Sciences,
3(4): 729-731.
Padhye AD and Ghate HV (2012) Fauna of Maharashtra, State
Faunal Series 20, Zoological survey of India, pp:239-246.
Padhye AD, Modak N, and Dahanukar N (2014) Indirana
chiravasi, a new species of leaping frog (Anura:
Ranixalidae) from Western Ghats of India. Journal of
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Padhye AD, Jadhav A, Modak N, Nameer D, and Dahanukar N
(2015) Hydrophylax bahuvistara, a new Species of
Fungoid frog (Amphibia: Ranidae) from Peninsular
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Padhye AD, Dahanukar N, Sulakhe S, Dandekar N, Limaye S
and Jamdade K (2017) Sphaerothica pashchima, a new
species of burrowing frog (Anura: Dicroglossidae) from

RESEARCH ARTICLE
Comparative Study of Antimicrobial Potentials of Phospholipid

Compound Produced by Halophilic and Alkaliphiles Bacillus
subtilis isolated from Alkaline Meteorite Crater
Lonar lake, India
Shinde VA1*, Patil RB2 and Pawar PV3
1Department of Microbiology, 2Department of Botany Shankarrao Chavan Mahavidyalaya, Ardhapur, Nanded,

Maharashtra, India.
3Department of Botany, Madhavrao Patil College, Palam, Hingoli, Maharashtra, India.
*Address for Correspondence : Email: shinde_vinod21@yahoo.com | Phone No. : +91-9921214222,

Received: 30.07.2017 Bacillus subtilis has ability to grow in extreme environments like
Accepted: 06.09.2017 Alkaliphilic, Halophilic. These environmental conditions induce
Published : 23.09.2017 microorganisms to produce varied kinds of antimicrobials which has
applications in chemotherapy. Therefore In present study antibiotic
Editor:
producing ability of the halophilic and alkaliphilic Bacillus subtilis were
Dr. Arvind Chavhan
compared by screening the activity of the antimicrobial compound was
Cite this article as: determined against different microorganisms like, Staphylococcus aureus, E.
Shinde VA, Patil RB and Pawar PV coli, P. aeruginosa, Candida tropicalis, Candida parapsilosis. The halophilic
(2017) Comparative Study of and alkaliphilic Bacillus subtilis isolates were isolated from Alkaline
Antimicrobial Potentials of Meteorite Crater Lake Lonar situated in district Buldhana, India and were
Phospholipid Compound Produced subjected to primary screening for antimicrobial compound production.
by Halophilic and Alkaliphiles The organisms were enriched in Alkaline and Halophilic NG medium which
Bacillus subtilis Isolated from
supports the production of phospholipid antibiotic by respective organism
Alkaline Meteorite Crater Lonar lake,
and partial purification was done using thin layer chromatography. The
India; International J. of Life Sciences,
5 (3): 420-424. partially purified extract of antimicrobial phospholipid compound from
both the organisms were tested against different microorganisms and seen
that halophilic Bacillus subtilis showed less antimicrobial activity than
Copyright: 2017| Author (s), This alkalophilic Bacillus subtilis isolated from Lonar Lake which are having
is an open access article under the moderate antibacterial activity against different microorganisms.
Attribution-Non-Commercial - No Keywords: Halophiles, Alkalophiles, Phospholipid antibiotic, Thin layer
Derivs License, which permits use chromatography, Antimicrobial Compound.
INTRODUCTION
Hypersaline environments are widely distributed on the earths continent
where they exist either as natural water bodies such as permanent saline
lakes, ephemeral saltpans and salt marshes, or as artificial solar salterns
(Litchfield and Gillevet, 2002). The microbial diversity of saline lakes has

Comparative Study of Antimicrobial Potentials of Phospholipid Compound
been studied primarily by focusing on the isolation Lonar lake was recorded. Same sample was used for
and characterization of individual organisms with further study.
potential industrial application (Horikoshi, 1999;
Jones et al, 1998). The spread of resistance to Media: Nutrient agar and broth was used for isolation
antibiotics undermines the therapeutic utility of Anti- of B. subtilis and production of antimicrobial
infective drugs in current clinical use (Bax et compound. Nutrient agar having pH 10 with 15% NaCl
al,2000). For example, Staphylococcus aureus, a major was used for cultivation of halophilic bacteria and
cause of community and hospital acquired infections, Nutrient agar having pH 10 was used for cultivation of
has developed resistance to most classes of alkaliphilic bacteria from Soil sample and was
antibiotics, and isolates exhibiting such resistance is incubated at 300C for 48 hrs.
drawing great concern. Methicillin - resistant Staph.
aureus (MRSA) strains appeared in the hospital Isolation and identification of bacteria: Nutrient
environment after introduction of the semi-synthetic agar having pH 10 with 15% NaCl & Nutrient agar
Penicillin, Methicillin, leaving Vancomycin as the having pH 10 were used for isolation of halophilic and
last line of defense for MRSA treatment (Enright, alkaliphilic Bacillus species respectively. Colonies
2003) Search for new antibiotics effective against showing characteristic feature were selected and
multi-drug resistant pathogenic bacteria is presently confirmed by colony character and biochemical test.
an important area of antibiotic research (Hamaki et al, These strains were selected for further study. Bergeys
2005). One strategy for enhancing the likelihood of manual of systematic bacteriology 9th edition was
obtaining novel antibiotic compounds and other followed for confirmation (Bergey et al, 1994).
secondary metabolites is to analyze uncommon
ecosystems which exist under extreme conditions Inoculum: Bacterial suspension was prepared by
(Okami and Hotta, 1988). adding 10 ml sterile water to a 4-day-old slant culture
and 5 ml of this was used as inoculum in all
A series of 167 peptide antibiotics have been recently experiments unless and otherwise stated. In each case
isolated from well-known Bacillus subtilis strains the bacterial suspension was standardized to have 0.5
(Berdy, 1974). The gram positive bacterium Bacillus O. D. at A600 (McFarland Standards). All experiments
subtilis produces a large number of antibiotics, which were conducted in the triplicates.
are classified as ribosomal or non-ribosomal. The non-
ribosomal antibiotics may play a role in competition Production of phospholipid antimicrobial
with other microorganisms during spore germination compound: Bacillus subtilis was grown in NG medium
(Tamehiro, 2002). adjusted to pH 10 (containing 10 gm (Gram) Nutrient
broth; 10 gm Glucose; 15% Sodium chloride; 5 mg
The phospholipid antibiotic produced by Bacillus (miligram) of CuSO4.5H2O; 7.5 mg of FeSO4.7H2O; 3.6
subtilis has the broad spectrum activity against gram gm of MnSO4.5H2O; 15 mg of CaCl2.2H2O; and 9 mg of
positive and gram negative bacteria. This study is ZnSO4.7H2O; (per liter)) supplemented with 50 g for
taken with the objective of isolation of halophilic and tryptophan per ml at 30oC for 24 hours (Tamehiro et
alkaliphilic Bacillus subtilis from the soil and al, 2002). 10% of this inoculum was reinoculated in
comparative study of antimicrobial potentials of fresh NG medium adjusted to pH 10 & fresh NG
phospholipid compound produced by halophilic and medium adjusted to pH 10 with 15% NaCl and
alkaliphilic Bacillus subtilis and activity was tested incubated under shaking at 300C for 72 Hours for
against test organisms (E. coli, Pseudomonas alkaliphilic and halophilic Bacillus subtilis respectively.
aeruginosa, Candida tropicalis, Staphylococcus Then these production medium were centrifugation at
aureus). 10000 rpm for 10 minutes, (Using Cooling centrifuge,
REMI) cells were collected. This cellular contents
were extracted three times using 10ml of 50% n-
MATERIAL AND METHODS butanol each time, then each aqueous layer were
collected and evaporated to concentrate at room
Collection of water sample: temperature (Bligh and Dyer, 1959). Resulting crude
Soil sample was collected from Lonar lake (Narayan et extract were resuspended in 4 ml of Methanol; this
al, 2008; Abou-Shanab, 2007). Temperature and pH of crude sample were again extracted with ethyl acetate.

Shinde et al., 2017
The resulting crude extract were used for further bioactive microbial metabolites by Bacillus subtilis.
purification. Purification was carried out using method Twenty five isolates from were isolated from
for lipid extraction (Katz and Demain, 1977). halophilic and alkalophilic culture medium and
identified according to Bergeys Manual of systematic
Bioassay of phospholipid antimicrobial compound: bacteriology (Bergey et al, 1994). These isolates were
The crude extract of antimicrobial compound with screened for antimicrobial activity of crude
ethyl acetate from halophilic and alkalophilic Bacillus phospholipid and three isolates from the halophilic
subtilis were used for bioassay. The 24 hours old and four isolates from the alkalophilic isolates showed
cultures of test organisms, Staphylococcus aureus, E. activity against a variety of organisms like,
coli, Pseudomonas aeruginosa, Candida tropicalis, and Staphylococcus aureus; E. coli, Pseudomonas
Candida Parapsilosis were streaked on sterile Muller aeruginosa, Candida tropicalis, and Candida
Hinton (MH) agar with sterile swabs. Then wells were parapsilosis but Halophilic isolates exhibited better
made on MH agar. The wells were filled with 100 l of activity against gram positive Staphylococcus aureus;
crude extracts of phospholipid compound. The plates Gram negative E. coli, Pseudomonas aeruginosa, and
were kept in refrigerator for diffusion of compound. Candida parapsilosis and weaker activity against
The plates were then incubated at 350.5 0 C for 24 Candida tropicalis and alkaliphilic isolates showed
hours and then diameter of zone of inhibition was moderate activity against gram positive
noted. Staphylococcus aureus; Gram negative E. coli, and
weaker activity against Pseudomonas aeruginosa,
Purification of phospholipid antimicrobial Candida parapsilosis and Candida tropicalis. Therefore,
compound: the isolates selected were used for phospholipid
The crude extracts were taken 1 ml and to it 3.75 ml antimicrobial compound production and the purified
1:2 (v/v) CHCl3: Methanol was added and vortex well. phospholipid was tested against Staphylococcus
Finally 1.25 ml distilled water was added and mix well aureus, E. coli, Pseudomonas aeruginosa, and Candida
and then centrifuged at 1000 rpm for 5 minutes at parapsilosis (Table.1).
room temperature, to give two phase system. The
bottom phase was removed and then TLC (Thin Layer In the search for antibiotics produced by Bacillus
Chromatography) was performed using silica gel. The species, especially Bacillus cereus, Bacillus subtilis and
plates were spotted with the bottom phase and plates Bacillus licheniformis, several antifungal compounds,
were developed with CHCl3 : Methanol: water mainly peptides, have also been described (Kugler et
(65:25:04 v/v). The phospholipid spots were located al, 1990; Lebbadi et al, 1994; Silo-Suh et al, 1994;
on chromatogram by placing the plates in iodine Bottone and Peluso, 2003). Some researchers have
chamber to treat with iodine vapour. After locating reported a compound produced by Bacillus pumilus
spots of Phospholipid antimicrobial compound from (MSH) that inhibits Mucoraceae and Aspergillus
halophilic and alkalophilic Bacillus subtilis, the spots species. Also different scientists have reported
were removed and extracted with CHCl3 : Methanol inhibition of various organisms (Maraheil et al, 1997).
(Katz and Demain, 1977). Some researchers isolated a strain of Bacillus subtilis
C126 from sugar cane fermentation, which produced a
Antimicrobial activity of purified phospholipid polypeptide antibiotic, bacitracin, which inhibited the
antimicrobial compound: growth of Micrococcus flavus (Tamehiro et al, 2002). A
The extracted purified phospholipid antimicrobial Bacillus licheniformis strain, 189, isolated from a hot
compound was filled in well prepared in MH agar spring environment in the Azores, Portugal, was found
plates inoculated with Staphylococcus aureus and to strongly inhibit growth of Gram-positive bacteria by
incubated at 350.5 C for 24 hours. After incubation producing peptide antibiotic (Mendo et al,2004). In
diameter of zone of inhibition was recorded. present study, the purified compound from halophilic
and alkaliphilic isolates were further purified by using
thin layer chromatography. The spot was detected and
RESULTS AND DISCUSSION the compound was fractionated. The antimicrobial
activity of the collected fraction was determined
The present research work was carried out to against most sensitive organism Staphylococcus aureus
optimize the conditions for the production of at 108 cells/ ml, and it was seen that the halophilic

Comparative Study of Antimicrobial Potentials of Phospholipid Compound
Table.1 Comparative Screening of Bacterial Isolates for phospholipid antimicrobial compound

Bacterial Isolates Antimicrobial activity of phospholipid antimicrobial compound
S. Aureus E. coli P. aeruginosa Candida
parapsilosis
Halophilic Isolates
BS 11 ++ ++ ++ ++
BS 18 ++ ++ ++ ++
BS 21 ++ ++ ++ ++
Alkaliphilic Isolates
BS1 ++ ++ + +
BS 2 ++ ++ + +
BS 7 ++ ++ + +
BS 8 ++ ++ + +
Table.2 Comparative study of Antimicrobial activity of phospholipid antimicrobial compound produced by

Halophilic and Alkaliphilic isolates.
Bacterial Isolates Test Organism Antimicrobial activity of phospholipid

antimicrobial compound produced by (in mm)
Halophilic Isolates
BS 11 Staph. aureus 25
Alkaliphilic Isolates
BS1 Staph. aureus 21
isolates exhibited better activity against Horikoshi K (1999) Alkaliphiles: Some applications of their
Staphylococcus aureus than that of alkalophilic isolates products for biotechnology. Microbiol. Mol. Biol. Rev., 63:
735-750.
(Table. 2). Multiplicity of antibiotic production
Jones BE, Grant WD, Duckworth AW and Owenson GG (1998)
obviously complicates attempts to identify antibiotics Microbial diversity of soda lakes. Extremophiles 2: 191-
in unfractionated material. However, even in whole 200.
cultures similarities or differences between known Bax R, Multan N, Verhoef (2000) The millennium bugs-the
and unidentified antibiotics may be noted by means of need for and development of new antibacterials. Int. J.
Antimicrob. Agents. 16: 51- 59.
chromatography (Snell et al, 1955).
Enright M (2003) The evolution of resistant pathogens-the
case of MRSA. Curr. Opin. Pharmacol. 3: 474-479.
Hamaki T, Suzuki M, Fudou R, Jojima Y, Kajiura T, Tabuchi A,
Sen K and Shibai H. (2005) Isolation of novel bacteria
interest. and actinomycetes using soil extract agar medium. J.
Biosci. Bioengineer. 99: 485-92.
Okami Y and Hotta K (1988) Search and discovery of new
REFERENCES antibiotics. In: Goodfellow M, Williams ST, Mordarski
M. (eds.) Actinomycetes in Biotechnology. Academic
Press, London, pp. 41.
Litchfield CD and Gillevet M P (2002) Microbial Diversity
And Complexity In Hypersaline Environments: A Berdy J (1974) Recent developments of antibiotic research
Preliminary Assessment. Ind. Microbiol.Biotechnol 28: and classification of antibiotics according to chemical
48-55. structure. Adv. Appl. Microbiol., 18: 309-406.
Norimasa Tamehiro (2002) Bacilysocin, a Novel
Phospholipid Antibiotic Produced by Bacillus subtilis

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315320
Narayan VV, Hatha MA, Morgan HW and Rao D (2008)
Isolation and characterization of aerobic thermophilic
bacteria from the Savusavu Hot spring in Fiji, Microbes
Environ., 23: 350-352.
Abou-Shanab R.A.I. (2007) Characterization and 16S rDNA
identification of thermo-tolerant bacteria isolated from
hot spring. J. appl. Sci. Res. 3:994-1000.
Bergey DH, Holt JG, Krieg NR and Sneath P.H.A. (1994)
Bergeys Mannual of Determinative Bacteriology, 9 th
ed., (Breed RS, Murray EGD and Smith NR, eds.)
Williams and Wilkins, Baltimore.
Tamehiro N, Okamoto-Hosoya Y, Okamoto S, Ubukata M,
Hamada M, Naganawa H. and Ochi K. (2002) Bacilysocin,
a novel phospholipid antibiotic produced by Bacillus
subtilis 168. Antimicrob. Agents Chemother., 46: 315-320
Bligh EG and Dyer WJ (1959) A rapid method for total lipid
extraction and purification, Can. J. Biochem. Physiol., 37:
911-917.
Katz E and Demain AL (1977) The peptide antibiotics of
Bacillus: chemistry, biogenesis, and possible functions.
Bacteriol. Rev., 41: 449-474.
Kugler M, Loeffler W, Rapp C, Kern A and Jung AG (1990)
Rhizocticin A, an antifungal phosphono-oligopeptide of
Bacillus subtilis ATCC 6633: biological properties. Arch.
Microbiol. 153: 276-281.
Lebbadi M, Galvez A, Maqueda M, Martinez-Bueno M and
Valihuia E (1994) Fungicin M4: a narrow spectrum
peptide antibiotic from Bacillus licheniformis M-4. J.
Appl. Bacteriol. 77: 49-53.
Silo-Suh LA, Lethbridge BJ, Raffel SJ, Clardy J and
Handelsman J (1994) Biological activities of two
fungistatic antibiotics produced by Bacillus cereus UW
85. Appl. Environ. Microbiol. 60: 2023-2030.
Bottone EJ and Peluso RW (2003) Production by Bacillus
pumilus(MSH) of an antifungal compound that is active
against Mucoraceae and Aspergillusspecies:
preliminary report. J. Med. Microbiol. 52: 69-74.
Maraheil MA, Stachelhaus T and Mootz HD (1997) Modular
peptide synthetases involved in non-ribosomal
pepetide syhtesis. Chem. Rev. 97: 2651-2673.
Tamehiro N, Okamoto-Hosoya Y, Okamoto S, Ubukata M,
Hamada M, Naganawa H and Ochi K (2002) Bacilysocin,
a novel phospholipid antibiotic produced by Bacillus
subtilis 168. Antimicrob. Agents Chemother. 46: 315-320.
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(2004) Purification and characterization of a new
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Snell N, Ijichi K and Lewis JC (1955) Paper Chromatographic
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Antibiotics. Western Utilization Research Branch, U.S
Department of Agriculture, Albany, California.

RESEARCH ARTICLE
Comparative study to evaluate in vivo and in vitro antioxidant

potential of processed and unprocessed Curcuma logna L.
using Allxon induced diabetic rats
Parashare VM1, Nikam S1* Nade VS 2, Pande Shishir3
1Department of Biochemistry, KTHM College, Gangapur Road, Nashik (MH), India

2Department of Pharmacognosy, MVPSs College of Pharmacy,Nashik (MH), India.
3Department of Rasshastra & B.K., ASS Ayurved Mahavidyalaya, Nashik (MH), India
*Corresponding author e-mail: parashare@gmail.com,

Received: 14.07.2017 The present study was aimed to investigate in vivo, in vitro antioxidant
Revised :16.08.2017 activity of the processed curcuma Longa L. turmeric (PT) and unprocessed
Accepted: 16.09.2017 curcuma Longa L. (UPT). Both the extract are subjected to qualitative
Published : 27.09.2017
phytochemical screening. In in-vitro antioxidant properties total Phenolic
content in each extract was determined using Gallic acid as a standard,
Editor:
Dr. Arvind Chavhan calibration curve was prepared. The result is expressed as mg of Gallic acid
equivalent per gram of extract. For unprocessed turmeric (UPT) and
Cite this article as: processed turmeric (PT) the values were 2.4 mg of GAE/g and 2.1mg of
Parashare VM, Nikam S Nade VS and GAE/g. respectively. In in-vitro antioxidant properties total flavonoid
Pande Shishir (2017) Comparative content was determined in each extract using Quercetin as reference
study to evaluate in vivo and in vitro standards. Total flavonoid content in methanolic extract of UPT and PT in
antioxidant potential of processed terms of quercetin equivalent (QE) was 152.25mg/100 g and 37.98mg/
and unprocessed Curcuma logna L.
100 g of dry extract respectively. Total antioxidant activity of UPT and PT
using Allxon induced diabetic rats;
extract in terms of ascorbic acid equivalent (AAE) was 6.394 and 3.922 mg/
425-437. ml of extract respectively.
In- vivo study involved five groups of each comprising six Wistar rats (250 -
Copyright: 2017| Author (s), This 300 kg) and labelled as Hyperglycemic, Normal, Standard drug Glipizide,
is an open access article under the UPT treated and PT treated. Besides the Normal group Alloxan was
terms of the Creative Commons administered in all animals as a single dose (50 mg/kg, i.v.) to induce
Attribution-Non-Commercial - No diabetes. A dose of 300 mg of processed and unprocessed turmeric/kg of
body weight was evaluated using acute toxicity test. It is orally
administered daily for four weeks after induction of diabetic. The fasting
properly cited, the use is non- blood glucose level was measured. The animal whose glucose level did not
commercial and no modifications or rise to more than 250mg/dl was rejected. Only animals that presented with
adaptations are made. glycemic levels equal to or above 250 mg/dl were submitted to treatments,
which consisted of a daily administration of the UPT and PT turmeric for
21days. It was considered as a first day and blood samples (Approximately
0.5ml) were collected from rats by puncturing the retro orbital sinus, under
mild ether anesthesia on 1st,7th, 14th 21st day for glucose estimations using
GOD-POD method. At the same time urine sugar was estimated using
Diastic. At the end of the study all groups rats pancreases were removed,

Parashare et al., 2017
labelled, were minced and 10% (W/V) tissue homogenate was prepared
using 0.1 M Phosphate buffer (PH 7.4) in a Potter- Elvehjem type
homogenizer and used to enzyme assay. The comparison using statistical
analysis ANOVA was performed. Decrease level of lipid peroxidase as
compare to Diabetic, increase in superoxide dismutase (SOD) and catalase
(CAT) activity are statistically analysed.( p<0.05).These findings suggest
that the UPT has potent antioxidant and antidiabetic activity, which may be
responsible for some of its reported pharmacological activities and can be
used as antioxidant supplement. Substances, chemicals rendering anti-
oxidative properties can attenuate alloxan toxicity.
Key words: Antioxidant, Unprocessed Turmeric (UPT), Processed

Turmeric ( PT), Diabetic Mellitus.(DM) Reactive oxygen species( ROS).
INTRODUCTION intracellular redox state destroys cell membranes and

kills cells (Kim, 2003). Thus, ROS are involved in the
Biological combustion involved in various processes pathogenesis of number of disease and aging process
produces harmful products or intermediates called (Mittler, 2007). Overproduction of ROS and/or
reactive oxygen species (ROS) or free radicals and thus inadequate antioxidants has been implicated in the
produced continuously in various metabolic processes pathogenesis and complications of some disease
and exist in biological systems. Superoxide (O2) is conditions like diabetes, Alzheimers disease, cancer,
the proximal mitochondrial ROS (Murphy, 2008). atherosclerosis, arthritis, neurodegenerative disease,
Some are important for maintaining normal and aging process. (Khalaf, 2008; Patel, 2010). Excess
physiological function. of free radicals in living beings has been known to
cause various problems like asthma, liver diseases,
ROS are produced naturally in cell following stress, muscular degeneration, and other inflammatory
respiration and exogenous sources of free radicals processes (Sen, 2010), resulting in the so-called
include tobacco smoke, ionizing radiation, certain oxidative stress. Oxidative stress is defined as
pollutants, organic solvents, pesticides, bacterial and imbalance between oxidants and antioxidants and
viral toxin, various smoking, alcohol, and psychological causes damage in to the cell and cell environment
or emotional stress etc. (Adeolu, 2009) Oxidative (Droge, 2002). Free radicals generated in the body can
stress is involved in the development and progression be removed by bodys own natural antioxidant defence
of diabetes- associated complications. In e.g. glutathione, catalase, SOD etc. but endogenous
hyperglycaemic condition, continuous generation of antioxidant defences are not completely efficient.
ROS occurs and the evidence showed diabetes induced Therefore, dietary supplement of antioxidant enzymes
changes in the activities of antioxidant enzymes in and free radical scavengers may provide a defensive
various tissues. mechanism against the deleterious acting of ROS, to
lower the effect of stress and complications of DM.
ROS are various forms of activated oxygen, free
radicals such as superoxide anion radicals (O-2) and Antioxidants have been reported to prevent oxidative
hydroxyl radicals (OH-1), as well as non-free radical damage caused by ROS by reacting with free radicals,
species (H2O2) and the singlet oxygen (1O2) chelating, and catalytic metals and also by acting as
(Halliwell, 1995; Squadriato and Peyor, 1998; Yildrim, oxygen scavengers (Shahidi, 1992). Antioxidants act as
2001; Gulcin, 2002). free radical scavengers, reducing agents, and
activators for antioxidative enzyme. These activities
The free radical may cause lipid peroxidation, are used to suppress the damage induced by free
aggregation of protein, Protein oxidation and the radicals in biological system. It has been found by
degradation of DNA, protein and polysaccharide (Esra, many researchers that there is an inverse association
2012) inactivation of enzymes and alteration of between the mortality from age related diseases and

In vivo and in vitro antioxidant potential of processed and unprocessed Curcuma logna L
the consumption of plant products. Because the plant intracellular reactive ROS, produced by the proton
products contain various antioxidant compounds, electromechanical gradient generated by the
phenolic, which are the most reactive compounds, mitochondrial electron transport chain and resulting
antioxidants present in plant products help in the in increased production of superoxide (Al-Shamaony,
stimulation of cellular defence system and biological 1994 ).
system against oxidative damage.
Turmeric (Curcuma longa L.) is a rhizomatous
The antioxidants in biological system can be either herbaceous perennial plant of the ginger family,
enzymatic or non-enzymatic. The enzymatic Zingiberaceae. It is native in southeast India, and
antioxidants include catalase, superoxide dismutase needs temperatures between 20C and 30C. When not
and glutathione etc. which catalyse neutralization of used fresh, the rhizomes are boiled for about 3045
many types of free radicals (Jacob, 1995). while the minutes and then dried in hot ovens (Chattopadhyay,
non-enzymatic antioxidants include Vitamin C, 2004). Some farmers use alkaline medium to boil the
selenium, vitamin E, carotenoids, and polyphenols etc. rhizomes, after which they are dried, polished used to
These enzymatic and non-enzymatic antioxidants can store or used to ground into a deep orange-yellow
terminate or prevent the formation of free radicals by powder. Turmeric has been used in Asia for thousands
donating hydrogen or electrons to reactive radicals or of years and is a major part of Siddha medicine
species. (Tayyem et al., 2004).
There is growing evidence that antioxidants play a

pivotal role in the prevention of heart disease, cancer, MATERIALS AND METHODS
DNA degeneration, pulmonary disease, and neurolo-
gical disorder. Plants, herbs, and spice, rich in phenolic Material:
compounds like flavonoids, have been demonstrated In the present study UPT and PT has been purchased
to have anti-inflammatory, antiallergenic, antiviral, from the same farmer. The UPT was washed and cut
antiaging, and anti-carcinogenic activities which can into a similar size pieces, sun dried and used further as
be attributed to their antioxidant properties (Ahmad, requirement. Alloxan and Nitroblue tetrazolium was
2006). purchased from Sigma Chemicals. (St. Louis Mo, USA)
and all remaining chemicals are of analytical grade and
Worldwide the major health problem is Diabetes were purchased from local distributors.
mellitus (DM) which includes insulin dependent
Diabetes mellitus (Type-1) and insulin independent Method, Experimental Design:
Diabetes mellitus (Type-2). (Gandhi 2017) Numerous
animal models have been developed for the past few Collection and authentication of plant material:
decades for studying diabetes mellitus and testing The Fresh Turmeric rhizomes (Curcuma Longa L.)
antidiabetic agents. were purchased from the Farmer of Rameshwar,
District Sangali, Maharashtra, during the month of
The standard method of inducing diabetes in animal April. The Sangali district is famous for the cultivation
models is with alloxan, a toxic glucose analogue, which and production of Turmeric. The rhizomes were
selectively destroys insulin-producing (beta) cells in washed using water. An herbarium of plant Curcuma
the pancreas when administered to rodents and many Longa L. was authenticated at the Botany research
other animal species. This causes an insulin-dependent office (Botanist) from the office of the Botanical survey
diabetes mellitus called alloxan-diabetes in these of India, Pune (MH) where a voucher specimen was
animals, with characteristics similar type 1 diabetes in submitted.
humans. The alloxan model of diabetes was first
described in rabbits (Dunn, 1943). Hyperglycaemia Preparation of drug and extract:
increases the production of reactive oxygen species The fresh rhizomes were cut into small and uniform
inside cultured bovine aortic endothelial cells. pieces and air-dried for 3 days in sunlight which are
(Nishikawa, 2000) Hyperglycemia can increase further used for extraction and also subjected to
oxidative stress through several pathways. A major pulverization to get coarse powder. The fine powder
mechanism appears to be the hyperglycemia-induced was kept in air tight glass containers. The complete

processed turmeric was also cleaned, cut into uniform C = (c x V)/m Where; C = total content of phenolic
pieces used for extraction and pulverised and store in compounds, mg/gm plant extract, in GAE, c = the
air tight glass container. Both containers are labelled concentration of gallic acid established from the
properly. calibration curve (mg/ml), V = the volume of extract in
ml, m = the weight of crude plant extract in gram.
Air dried UPT was extracted in methanol for 72 hours
using Soxhlet apparatus. The extract was concentrated Quantification of total flavonoids:
under reduced pressure using a rotator evaporator at Total flavonoid content was measured by the
40C until the solvent completely dried. Turmeric is Aluminium chloride colorimetric assay (Zhishen,
insoluble in water. The yield of the water extract for 1999). Quercetin was used as a standard and total
unprocessed turmeric was 7% and of processed flavonoid content of extracts are expressed as mg
turmeric was 4.5%. The extract was and stored in air quercetin equivalents (QE)/100 g of sample. An
tight container at 40C which is used for further aliquot of 1 ml of extracts and standard solution of
studies. Processed and unprocessed turmeric powder Quercetin (0.1mg /ml) was added to 10 ml volumetric
mercerized in the carboxy- Methyl Cellulose (CMC) flask containing 4 ml of distilled water. To this 0.3 ml 5
Sodium Salt was used as a drug and administered % NaNO2 were added. After 5 min, 0.3 ml 10 % AlCl3
orally by intra-gastric intubation. was added. Then at the 5th min, 2ml of 1 M NaOH was
added and the total volume was made up to 10 ml with
Preliminary phytochemical analysis: distilled water. The solution was mixed well and the
Test solution was prepared by dissolving 1.5 gram of absorbance was measured against prepared reagent
both extract separately in 15 ml distilled water. Each blank at 510nm. Calibration curve of quercetin was
extract was tested for the presence of different phyto- prepared using aliquots of 0.2, 0.4, 0.6, 0.8, 1, 1.2, 1.4,
constituents, viz. alkaloids, Carbohydrates, steroids, 1.6, 1.8, and 2 ml standard solution. Accordingly,
glycosides, phenols, flavonoids, saponins, proportionate additions were done as describe above.
triterpenoids, Phlobatanins, cardiac glucosides, The solution was mixed well and the absorbance was
Anthraquinone, proteins by usual prescribe methods measured against prepared reagent blank at
(Kokte, 2004; Harborne, 2006). 510nm.The calibration curve was plotted. A blank,
which was prepared by the same procedure except
Quantification of total phenolic: replacing the plant extract with an equal volume of
The concentration of total phenolic content in the both methanol. The total content of flavonoid compounds in
extract was determined by the Folin-Ciocalteu plant methanol extracts in quercetin equivalents was
colorimetric method (Singleton, 1965; Sadashivan, calculated by the following equation:
1997). Gallic acid was used as a standard. Calibration
curve is prepared using dilution of standard solution C = (c x V)/m Where; C = total content of flavonoid
of 0.1mg/ml of Gallic acid. Samples of extracts were compounds, mg/gm plant extract, in quercetin
evaluated at a final concentration of 0.1 mg/ml and equivalent, c = the concentration of quercetin
were mixed thoroughly with 2 ml of 2% Na2CO3. After established from the calibration curve in mg/ml, V =
2 minutes Folin-Ciocalteu reagent was added to the the volume of extract in ml and m = the weight of
mixture. The resulting mixture was allowed to stand at crude plant extract in gm.
room temperature for 30 min and the absorbance was
measured at 743 nm against a blank. Total phenolic Reducing Power Capacity Assessment:
content was expressed as gram of Gallic equivalents The reducing power of UPT and PT was determined by
per 100 grams of dry weight (g 100g-1D.W.) of the the slight modification of the method of Oyaizu
plant extract. A blank was prepared using the same (Oyaizu, 1986). Substances, which have reduction
procedure but replacing the plant extract with an potential, react with potassium ferricyanide (Fe3+) to
equal volume of methanol. The total content of form potassium ferrocyanide (Fe2+), which then
phenolic compounds in plant extracts as gallic acid reacts with ferric chloride to form ferric ferrous
equivalents (GAE) was calculated using the following complex, having maximum absorption spectra at
equation (Demiray, 2009): 700nm. Various concentrations of the plant extracts
dissolved in water were mixed with 2.5 ml of
phosphate buffer and 2.5 ml of potassium ferricyanide.

This mixture was kept at 50C in water bath for 20 OECD guidelines, draft guidelines 423 adopted on 17
minutes. After cooling, 2.5 ml of 10% trichloroacetic December 2001 received from CPCSEA, Ministry of
acid was added and centrifuged at 3000 rpm for 10 Social Justice and Empowerment, Government of India.
min. The upper layer of solution (2.5 ml) was mixed The animals of either sex were segregated into six
with distilled water (2.5 ml) and a freshly prepared groups consisting of six rats each and were fasted for
ferric-chloride solution (0.5 ml). Control was prepared 18 hours and then use. There was an orally
in similar manner excluding samples. Stock standard administration of the stepwise doses in which the
solution of 1% ascorbic acid was prepared. Serial starting dose of unprocessed turmeric was 100mg/kg
dilutions of standard ascorbic acid 1% were used and of animal weight. The increasing doses were
calibration curve was prepared. Increases absorbance administered up-to 3000mg/kg of animal weight in a
of the reaction mixture containing extract at various range of 100mg/kg. Additionally, 3 mice were kept as
concentrations indicates increase in reducing power. control. Same procedure was repeated for processed
Reducing power was measured by varying the turmeric. Since no mortality was observed in both
concentration of the extract. The experiment was studies, the behavioral pattern was unaffected. Hence,
performed thrice and results were averaged. the dose selected for all the extract were as per the
OECD guidelines No 423, fixed dose methods for
In-vivo studies: evaluation of anti-diabetic activity was 300mg/kg,
Venues body weight (1/10 of 3000 mg/kg body weight.)
The whole experimental work was conducted at the
NDMVPs College of Pharmacy, the University of Pune. Grouping of animals with induction of diabetic
mellitus using alloxan:
Animals In this method, experimental rats were fasted in
Forty albino Wistar rats (190-220 g) of either sex were individual cage for 24 hours. Care was taken to avoid
employed in this study. They were housed in well carophagy. Forty Wister rats of either sex weighing
ventilated cages and maintained under standard 190 -220 gm were divided into five groups of six
laboratory conditions at 25 2C, relative humidity 50 animals each and grouped as follows [OECD/OCDE,
15% and normal photo period [12 h dark/12 h light]. OECD Guidelines 2000-2001].
They were used for the experiment. The animals were
maintained in accordance with the guidelines of the Group I:
National Institute of Nutrition, Hyderabad, India. The Served as normal control and did not receive any
rats were acclimatized for seven days in the laboratory treatment.
and were fed with Commercial pellet rat chow diet Group II:
(Hindustan lever, Kolkotta, India) and water was Served as diabetic and received single dose of alloxan
provided ad libitum. All the animal experiments were monohydrate (50mg/kg,i.p.)
conducted according to the ethical norms Group III
(OECD/OCDE, OECD Guidelines for the testing of Alloxan monohydrate + unprocessed turmeric (300
chemicals, revised draft guidelines 423:2001) mg/kg, p.o.)
approved by CPCSEA, Ministry of Social Justice and Group IV:
Empowerment, Government of India, and ethical Alloxan monohydrate + processed turmeric (300
clearance (IAEC/2014/14) was granted by mg/kg, p.o.)
Institutional Ethical Committee presentations held on Group V:
24\02\2014 at NDMVP Samajs College of Pharmacy, Alloxan monohydrate + Glipizide (5mg/kg, p.o.) and
Nashik. served as Standard.
Preliminary toxicological evaluation: Acute Diabetes was induced in the rat by injecting alloxan
toxicity test monohydrate intra-peritoneally in a single dose of
Acute toxicity is the adverse effect occurring within a 50mg/kg in 0.95% of weight/volume NaCI, to the
short time of oral or intra peritoneal administration of overnight fasted rat. Diabetes was confirmed in the
single dose of any substance given within 24 hours. alloxan treated rats by measuring the fasting blood
The acute oral toxicity studies of processed and glucose concentration 24 hrs-48 hrs (1-2 days) post
unprocessed turmeric were carried out as per the injection. (Parashare, 2016)

The fasting blood glucose level was measured. The estimation by GOD-POD method (Trinder, 1969). At
animal whose glucose level did not rise to more than the same time one drop of urine is used for glucose
250mg/dl was rejected. Only animals that presented assessment using Di-strips. Both these studies are
with glycaemic levels equal to or above 250 mg/dl carried on 1st, 7th, 14th, and 21st day of the study.
were submitted to treatments.
Determination of in vivo antioxidant activity:
Methodology of oral drug administration:
Intraperitoneal route of administration of drug is Estimation of antioxidant enzymes:
selected because it is more rapid and predictable Catalase activity was determined according to the
absorption than oral administration. This route is also method of luck (Luck, 1971). 3ml H2O2- phosphate
advantageous because the drug is not inactivated or buffer of pH 7 (12.5mM H2O2 in 67 mM Phosphate
destroyed as may happen in the gastrointestinal tract. buffer) was pipetted directly in the cuvette. 0.05 ml of
Rat oral gavage (16 gauze) needle was used, it was tissue homogenate (10%) mitochondrial supernatant
blunted at the tip to avoid any injury to the inner was added to the buffer solution. The content was
surface of mouth. The feeding needle was attached to mixed and the decomposition rate of hydrogen
the syringe. Required amount of drug solution was peroxide, decrease in the absorbance was followed
withdrawn in the syringe without any air bubble. By and noted at 240nm for 5 minutes. The results were
adjusting the needle to desire position it was inserted expressed as micromole of H2O2 decomposed/
through the intra-dental to space and pushed gently min/mg protein using Molar extinction coefficient of
(Book 1) . Study consisted of a daily administration of H2O2 (0.07 mM/cm) at 240nm. The enzyme solution
the PT and UPT for 21days.First drug administration containing H2O2-free phosphate buffer served as
day was considered as a first day. control. One enzyme unit was calculated as the amount
of enzyme required to decrease the absorbance at
Collection of Blood and Preparation of tissue 240nm by 0.05 units.
homogenate:
The blood samples (Approximately 1ml) were Superoxide dismutase activity was assayed by the
collected on 1st, 7th, 14th, and 21st day by puncturing method of Kono (Kono, 1978). An aqueous solution of
the retro orbital sinus, under mild ether anaesthesia in hydroxylamine was prepared daily and its pH was
a properly labelled EDTA bulb for glucose estimations adjusted to 6.0 with 2M sodium hydroxide. Control
using GOD-POD method. At the end of the study, 21st consist of a mix of 2ml of NBT (Nitroblue tetrazolium
day all animals were anesthetized in an ether chamber. 1.5mM)), 0.5ml hydroxylamine hydrochloride. The
The pancreas was removed and placed at 4C after reduction of NBT was inhibited by SOD and was
perfusion with ice cold saline with proper labelling. measured spectrophotometrically at 560nm. In a test,
Pancreas tissues from each animal were minced and enzymatic reaction, 2ml of NBT was mixed with 0.5ml
10% (W/V) tissue homogenate was prepared using 0.1 of hydroxylamine hydrochloride and appropriate
M Phosphate buffer (PH 7.4) in a Potter- Elvehjem type amount of mitochondrial supernatant was added the
homogenizer (Arulselvan, 2007). All the test tubes development of blue colour was measured at 560nm
were labelled carefully and then subjected to for 2 minutes. Enzyme activity was expressed as Units
centrifugation at 1000 g, 40C for 10 minutes. A pellet /mg protein where one unit of enzyme is defined as
from all the test tube was discarded. The supernatants the amount of SOD requires inhibiting rate of reaction
were subjected for centrifugation at 12.000 g for 20 by 50%.
minutes at 4oC to obtained mitochondrial supernatant
(PMS). The supernatants were sieved separately Estimation of lipid peroxidation (Niedernhofer et al.,
through two layers of muslin cloth and immediately 2003, Tietz, 1968) in terms of malonaldehyde (MA)
used to estimate the activity of catalase and thiobarbituric acid reaction was performed. The
superoxide dismutase (SOD), and lipid peroxidase. reaction mixture contained 0.1ml of tissue
homogenate, 0.2ml of 8.1% sodium dodecyl sulphate
Estimation of blood and urine glucose: (SDS), 1.5ml of 20% acetic acid and 1.5ml of 08%
The EDTA collected blood samples were centrifuge at aqueous solution of thiobarbituric acid (TBA). The pH
5000 RPM for 20 minutes to separates the plasma. of 20% acetic acid was adjusted with 1 NaOH to 3.5.
Plasma is immediately subjected for the glucose The mixture was finally made up to 40 ml with

distilled water and heated at 95C for 60 min. After These phytochemical compounds are known to
cooling under tap water, 1ml of distilled water and 5ml provide support for bioactive properties of plant, and
of a mixture of n-butanol and pyridine (15:1 by vol.) thus they are responsible for the antioxidant
was added and the mixture was shaken vigorously on properties of unprocessed curcuma Longa L.
a vortex mixer. After centrifugation at 2200 g for 5
minutes the absorbance of the organic layer (upper The total reducing power capacity using potassium
layer) was measured immediately at 532 nm. Lipid ferric cyanide reduction method is determined. In this
peroxide formation in tissues clearly requires assay, the yellow colour formed in the reaction is
substrates, the unsaturated fatty acids of the tissue significant indicator of antioxidant activity. The UPT
lipids, and a catalytic system. Either of these factors extract shows high absorbance of 1.48, then comes the
may limit or control the extent of peroxide formation. absorbance of PT 0.91. Fig. 3. Among the extract the
Malonaldehyde, an index of LPO was calculated with UPT extract exhibited the most reducing power. This
extinction coefficient of 1.5 x 105 M-1cm-1. result indicates that the extracts may consist of
polyphenolic compounds that usually show great
Statistical analysis reducing power. The reducing ability of a compound
The experimental results were calculated as mean generally depends on the presence of reductants
SD of six replicates. The ANOVA test followed by which have been exhibited antioxidative potential by
Dunnett's multiple comparison tests was employed for breaking the free radical chain, donating a hydrogen
statistical comparison between control and various atom (Gordon, 1990) The reducing capacity of UPT is a
groups. Significance was considered at P<0.05. significant indicator of its potential antioxidant
activity.
Results of the effect of unprocessed and processed

RESULTS AND DISCUSSIONS turmeric on blood glucose level of Alloxan -induced
diabetic rats are presented in Table 2. Significant
The phytochemical spot test showed that the extract antihyperglycemic effect was observed from the 7th
contained saponins, phenols, terpenes/ sterols, glycol- day onwards. The reduction in blood glucose was
sides, alkaloids, and flavonoids as given in Table 1. maximum on the fourth week, receiving 300mg/kg
body weight of unprocessed turmeric and the results
The total phenolic contents of extract of UPT and PT were comparable with that of glipizide a standard.
were found to be 16.5mg GAE/100g and 8.86 mg GAE Alloxan caused a significant increase in blood glucose
/100g respectively with reference to standard curve level as reported in the Table 2. The results of the
(y= 0.01284 X - 0.01533; R2 =0.9889) Fig. 1. The present study clearly indicated that the UPT exhibited
Literature reveals that antioxidant activity of plant significant antidiabetic activity in Alloxan induced-
extract is mainly due to presence of phenolic diabetic rats, comparable to the effect exhibited by
compounds, which may exert antioxidant effects as standard drug glipizide. The effect of administration of
free radical scavengers, as hydrogen donating sources UPT was found to reduce glucose concentration.
or as singlet oxygen quenchers and metal ion (***P 0.001). At the end of study after 21st days of
chelators. treatment period, it was observed that the animals
treated with UPT, PT and glipizide showed significant
Total flavonoid contents were calculated using the decrease in diabetes induced urine glucose level Table
standard curve of quercetin (y=0.5804X - 0.1058; R2= 3.
8654) "Fig. 2", and the result was expressed as
quercetin equivalents (QE) per 100 grams of the plant In present study the MDA level was found to be
extract. The total flavonoid contents of both significantly decrease in UPT and PT treated group of
unprocessed and processed turmeric extract were animals, as compare to diabetic group(P<0.001). The
found to be 75.625 mg QE/100g and 18.99 mg antioxidants SOD and Catalase were found to be lowest
QE/100g, respectively. According to our study, the in diabetic rats. The P value calculated for the rise in
high contents of flavonoids in unprocessed turmeric SOD, Catalase and lowering of MDA against the groups
extract can explain its high radical scavenging activity. as listed in Table 4 were all significant.

Table 1: Qualitative Phytochemical Screening of methanolic extract of UPT and PT.

Sr. Test Unprocessed turmeric Processed turmeric (PT) Inference
No (UPT) extract extract
1 Alkaloids
a Mayers test Yellow colour with Red colour with ppt. was Alkaloids
Brownish red ppt. was observed observed present
b Wagners test Yellow colour with Red colour with ppt. was Alkaloids
Brownish red ppt was observed observed present
c Hagers test Dark yellow colour was Red colour with ppt. was Alkaloids
observes observed present
2 Glycosides
a Benedicts test Orange red precipitate was Orange red precipitate was Glycosides
obtained obtained /reducing sugar
present
b Fehlings test First yellow and then brick red First yellow and then brick Glycosides
precipitate was obtained red precipitate was /reducing sugar
obtained present
3 Cardiac glycosides
a For deoxy sugars No brown ring at the junction of No brown ring at the Deoxysugars absent
Keller-killiani test two liquid, blackish colours is junction of two liquid,
formed. blackish colour formed.
b Legals test Negative, no pink, red, reddish Negative, no pink, red, Cardiac glycosides
brown colour, Dark yellow reddish brown colour, Dark absent
colour is formed. yellow colour is formed
4 Anthraquinone glucosides
a Borntragers test In the lower ammonical layer In the lower ammonical layer Absent
colour is absent colour is absent
5 Saponin glycoside
a Foam test slight foaming Slight foaming Saponins present
6 Flavonoids
a Alkaline Dark Yellow colour appear Yellow colour appear Flavonoids present
reagent test
b Shinoda test Orange colour appears Orange colour appears Flavonoids present
c Ammonia test Dark yellowish red color appear Dark yellow colour appear Flavonoids present
7 Phenolic compound
a Lead acetate test Clear dark yellow colour was Clear yellow colour was Phenols Present
observed with colourless observed with colourless
precipitate at bottom precipitate at bottom
8 Sterols
a Salkowski Dark red colour appears in the Dark yellow colour appear in Sterols present
reaction chloroform layer the chloroform layer
9 Proteins
a Biuret test No purple dark purple colour , No purple dark purple colour Protein absent
instead red colour appears , instead red colour appears
b Lowry reagent No purple dark purple colour , No purple dark purple colour Protein absent
instead red colour appears , instead red colour appears
b Ninhydrin test No deep violet colour appears No deep violet colour Protein absent
appears

Fig. 1 : Calibration curve of phenolic content using Fig. 2 : Calibration curve of Total flavonoid using
gallic acid as a standard quercetin as a standard
Fig. 3: Comparative reducing power capacity of Fig. 4: Effect of unprocessed and processed turmeric on
standards ascorbic acid, unprocessed and processed blood glucose on different groups: a) Group II compare
curcuma longa L with group b) Group III compare with group II c) Group
IV compare with group II d) Group V compare with
group II e) Group V compare with group III ns= Non
significant. ***P 0.001, **P 0.001, *P 0.05
Table 2: Effect of unprocessed and processed turmeric on the blood glucose level in Alloxan induced
diabetic rats.
Group Fasting blood glucose concentration Mg/dl
Day 1 Day 7 Day 14 Day 21
Normal Group I 75.00 0.706 77.49 1.600 75.11 1.716 74.40 0.988
Diabetic induced Group II 378.29 0.3898 355.9 0.2367 392.2 0.825 396.1 0.5345
Diabetic + UPT Group III 358.00 0.3122 242.41 1.736 185.5 0.9521 139.16 0.4771
Diabetic +PT Group IV 359.2 0.1983 317.4 0.7783 275.00 0.670 196.4 0.4666
Diabetic control Group V 359.5 0.1611 218.85 0.485 172.21 0.761 134.6 0.667
Values are expressed as Mean SD ( n =6 rats). The data of 21 days in all group of rats was subjected to ANNOVA.

Table 3: Effect of processed and unprocessed turmeric on the urine glucose level in Alloxan induced
diabetic rats.
Treatment Intensity of glucose in the urine before treamnent and from the 1 st day of treatment
0 day 1 day 7 day 14 day 21 day
Normal group I Nil Nil Nil Nil Nil
Diabetic induced group II Nil +++ ++++ ++++ ++++
Diabetic +UPT Nil +++ ++++ ++ ++
Diabetic +PT Nil +++ ++++ +++ ++
Diabetic + gilpizide Nil +++ ++++ ++ +
Intensity of glucose in urine, +: mild, ++: moderate, +++: higher, ++++: severe
Table 4: The antioxidant enzymes Superoxide dismutase (SOD), Lipid peroxidase(LP) , Catalase studied in
the pancrease tissue of all group animals at end of 21 st day.
Group Catalase SOD Lipid peroxidase
Normal Group I 12.02 0.2635 5.7650 0.2645 9.400 0.2978
Diabetic induced Group II 4.0855 0.1287# 1.810 0.03011# 26.840.4229 #
Diabetic + UPT Group III 9.970 0.04655a *** f *** 3.978 0.04527b ***e *** 17.7420.412b*** d ***
Diabetic + PT Group IV 7.82 0.09878a *** 2.667 1.535b *** 22.42 0.3555 b***
Diabetic+ Glipizide Group V 4.048 0.03414 b a *** 1.978 0.003 b*** 19.660.477 b***
Values are expressed as mean SD. (n=6) & are statistically significant at *P < 0.05, **P < 0.001 and ***P < 0.01
Fig. 5 Effect of UPT and PT on the

SOD, Catalase, LPO level in all
groups of studies. Values are the
mean SD for 6 animals in each
group. Values are statistically
significant at* P<0.05, **P<0.01,
P<0.001. Statistically significant
was compared within the groups
as:
aDiabetic rats compared with
Normal rats,
bGlipizide, UP and PT treated data
compared with Diabetic data.

Statistical significance was compared within the group some of which include the inhibition of reactive
as follows, oxygen species (ROS) generation, direct or indirect
a= catalase: Group II compare with the groups of UPT scavenging of free radicals, and alteration of
(III), PT (IV) and Glipizide (V) treatment. intracellular redox potential (Ekrem, 2008).
b= SOD: Group II compare with the groups of UPT (III), Antioxidants have functioned to inhibit apoptosis
PT (IV) and Glipizide (V) treatment. because apoptosis was at first thought to be mediated
c = LPO: Group II compare with the groups of UPT (III), by oxidative stress (Hockenbery, 1993). Crude extracts
PT (IV) and Glipizide (V) treatment of fruits, herbs, vegetables, cereals, and other plant
d= LPO: Group III (UPT) compare with group IV (PT) materials rich in phenolic are increasingly of interest
e = SOD: Group III (UPT) compare with group IV (PT) in the food industry because they retard oxidative
f=Catalase: Group III (UPT) compare with group IV degradation of lipids and thereby improve the quality
(PT). and nutritional value of food. The importance of the
antioxidant constituent of plant materials in the
Fig. 5 shows the level of catalase, SOD and LPO. It was maintenance of health and protection from coronary
observed that due to diabetes there was an increase in heart disease and cancer is also raising interest among
the MDA (LPO) and decrease in SOD and Catalase scientists, food manufacturers, and consumers as the
enzyme level. In diabetic glipizide treated rats the level trend of the future is moving toward functional food
of LPO is significantly decreases as compare to diabetic with specific health effects (Lo-liger, 1991). By taking
group. Treatment with UPT and PT reversed the MDA into consideration, the important of antioxidant UPT is
(LPO) level which is significantly decrease ( < 0.001) a good source.
and increase in SOD and catalase HDL level were
observed after the treatment with UPT and PT. ( < In the present study the total phenolic content of UPT
0.01). There is also a significant difference in the was 16.5mg GAE/100g and of PT was 8.86 mg
comparison of UPT and PT values. ( < 0.001). GAE/100g. Many studies focused on the biological
activities of phenolic compounds, which have potential
antioxidants and free radical scavengers. The
DISCUSSION antioxidant activity of phenolic is mainly due to their
redox properties, which allow them to act as reducing
Ethnopharmacological surveys have shed light on the agents, hydrogen donators, and singlet oxygen
fact that the therapeutic use of even 80% of 122 plant quenchers. In addition, they have a metal chelation
derived drugs may have a link with their potential. The phenolic compounds has been shown to
recommendations in traditional medicine (Fabricant, scavenge free radicals, including hydroxyl and
2001). Potential sources of antioxidant compounds superoxide anions (Noda et al., 1999 ). This reduces
have been searched in several types of plant materials the further complications of diabetic mellitus (Da Silva,
such as vegetables, fruits, leaves, oilseeds, cereal crops, 2010)
barks and roots, spices and herbs, and crude plant
drugs. (Ramratnam et al., 1995) Antioxidants (free Flavonoids are important secondary metabolites of
radical scavengers) are chemicals that interact with plant modulating lipid peroxidation involved in
and neutralize free radicals, thus preventing them atherogenesis, thrombosis and carcinogenesis. It has
from causing cellular damage in the biological system been confirmed that pharmacological effects of
(Diplock, et al., 1998). Besides endogenous flavonoids are correlating with their antioxidant
antioxidants such as SOD, catalase, Glutathione etc. the activities as radical scavenging activities and are very
body also relies on exogenous the dietary antioxidants, important due to the deleterious role of free radicals in
to obtain the rest of the antioxidants it needs such as biological systems (Mbaebie, 2012). In this study, it is
Vit. C, Vit. E etc. (Valko et al., 2007). The data obtained evident that the extract of the UPT possess more
on the basis of calibration curve highlighted the effective antioxidant activity than the PT. The
antioxidant potential of UPT. UPT is a best, and UPT is increased activities of catalase and SOD as observed in
cheap source of exogenous dietary antioxidant. In the this study suggest that the UPT extract has an in vivo
Indian kitchen Daily use of UPT instead of PT may antioxidant activity. The radical scavenging activity of
found to be beneficial. These exogenous natural SOD is effective only when it is followed by increase in
antioxidants have a diversity of biochemical activities, activity of catalase and other peroxidases. In our study,

the level of MDA in the extract treated groups Squadriato GL, Peyor WA (1998) Oxidative chemistry of
decreased in a dose dependent manner when nitric oxide: the role of superoxide,peroxynitrite, and
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compared to control. This decrease in the MDA levels
Yildrim A, Oktay M and Bilaloglu V (2001) The antioxidant
may indicate increase in the activities of glutathione activity of the leaves of Cydonia vulgaris. Turkish J.
peroxidase and hence inactivation of LPO reactions Med. Sc. 31:, 23-27.
(Ugochukwu, 2003). UPT dose level 300mg/kg of body Gulcin I, Oktay M, Kufrevioglu IO and Aslan A (2002)
weight shows significant low levels of blood glucose Determination of antioxidant activity of Lichen Cetraria
islandica (L), Ach. J. Ethnopharmacol. 79: 325-329.
which simultaneously proved by urine glucose level.
Esra Birben, Umit Murat Sahiner, Cansin Sackesen et al.,
Superoxide dismutase, catalase and non-enzymatic
(2012) Oxidative stress and Antioxidanr defence.
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Jacob RA (1995) The integrated antioxidant system,
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higher antioxidant potential and antihyperglycemic
Dunn JS, Sheehan HL, Mcletchie NGB (1943) Necrosis of
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RESEARCH ARTICLE
Studies on Physico-chemical analysis of Mohabala Lake near

Bhadrawati, District - Chandrapur (MS), India
Mahajan VS1 and Pokale SS2
1Department of Zoology, Centre for Higher Learning and Research in Zoology, Nilkanthrao Shinde Science and Arts
College, Bhadrawati - 442902, Dist- Chandrapur (M.S.), India.
2Dr. B. N. Purandare Arts, Smt. S. G. Gupta Commerce & Science College Lonavla.
Email: badkhalvm@gmail.com
Received: 26.05.2017 The Mohabala lake is principal fresh water body located within Bhadrawati
Accepted: 29.06.2017 tahsil in Chandrapur District of Maharashtra state. Now a day lakes are
Published : 27.09.2017 degraded by both natural and anthropogenic activities, which deteriorate
their quality of lake water. Normally, lakes perform the functions directly
Editor: related to their physical, chemical and biological integrity to decide quality
Dr. Arvind Chavhan status of water. The analysis of various physico-chemical characteristics of
water quality of Mohobala lake at different sites under study viz. Site A,
Site B and Site C were carried out for two years i.e. from June 2013 to May
Mahajan VS and Pokale (2017)
2015 for the detection of level of pollution, physic-chemical parameters
Studies on physico-chemical analysis
of Mohabala Lake near Bhadrawati, analysis like, Atmospheric temperature (C), Water temperature (C),
District - Chandrapur (MS), India; Transparency (cm.), Conductivity (mhos/cm.), pH, D.O. (mg/lit.), Free CO 2
International J. of Life Sciences, 5 (3): (mg/lit.), Total alkalinity (mg/lit.), Phenolphthalein alkalinity (mg/lit.),
438-446. Total hardness (mg/lit.), Calcium hardness (mg/lit.), Magnesium hardness
(mg/lit.), Chloride (mg/lit.), B.O.D. (mg/lit.), C.O.D. (mg/lit.), Phosphate
(mg/lit.), Sulphate (mg/lit.), Nitrate (mg/lit), Total solids (mg/lit.). Total
dissolved solids (mg/lit.), Total suspended solids (mg/lit.).
Copyright: 2017 | Author (s), This
is an open access article under the Key words: Physico-chemical, Analysis, Mohabala lake.
and distribution in any medium, INTRODUCTION
properly cited, the use is non- Water is the prime requirement for the existence of life and thus it has been
commercial and no modifications or mans endeavor for the time immemorial to utilize the available resources.
adaptations are made. The unbridled exploitation of water for irrigation, drinking and industrial
purposes has caused a drastic decline of the quality and availability of
water. Lakes, rivers and reservoirs are the very important water resources
and used for various purposes. Physico-chemical parameters are the
important component of the aquatic system as they indicate the water
quality of aquatic ecosystem. This present study was conducted at three
different sites in the Mohobala lake. Monthly and Seasonal variations in the
physico-chemical parameters in Mohobala lake were studied during the
study period of June 2013 to May 2015. This study indicates that the lakes
of central India exhibit substantial variation in their biotic and abiotic
characteristics

Studies on physico-chemical analysis of Mohabala Lake near Bhadrawati
MATERIAL AND METHODS of Open Reslus Method of APHA (1985) using Ferroin
indicator. Phosphate is measured by turbidometric
The water samples were collected from the three sites method with the help of Spectrophotometer at 690 nm
of Mohabala lake and name them site A site B and against a standard curve drawn from different known
site C. Marginal areas at 1 to 1.5 m depth with the concentrations. Sulphate was measured
help of Ruffners sampler. The water samples were Spectrophotometrically at 420 nm against a standard
collected in dried plastic cans of 5 lt. capacity. The curve drawn from different known concentrations.
water sample collected during the morning hours Nitrate was measured by Spectrophotometer by
between 8.30 to 10.30 am. The samples were collected cadmium reduction colorimetric method at 410 nm
each month for the period of 2 years i.e. from June against a standard curve drawn from different known
2013 to May 2015. The recorded data was separated in concentrations. APHA (1985). Total solids are
3 seasons, Monsoon (June to September), Winter determined as the residue left after evaporation of the
(October to January) and Summer (February to May). unfiltered sample. Total dissolved are determined as
During the determination, physicochemical parameter the residue left after evaporation of the filtered
like atmospheric temperature, water temperature, pH sample. Total Suspended Solids are determining as the
and transparency were determined on the sampling difference between the total solids and the total
spot. dissolved solids.
The mercury thermometer was employed to record

the temperature of the Atmosphere Temperature and RESULT AND DISCUSSION
for Water temperature. Transparency was determined
with the help of Secchi disc (Secchi 1865). The The atmospheric temperature was always found
conductivity of water was measured directly with the higher than the temperature of water. In the present
help of Conductivity Meter. The instrument (model no. investigation the atmospheric temperature was
EQ- 660) was checked constantly by using standard recorded is higher than the water temperature at all
potassium chloride solutions. Cell constant of the sampling three sites. It is fluctuated between 21.11C
conductance 822 mho cm-1.Total solids are determined to 45.28C in site A, 21.56C to 45.23C in site B and
as the residue left after evaporation of the unfiltered 21.15C to 45.21C in site C. Similar findings were also
sample. Total dissolved are determined as the residue recorded by Rawat and Jakher (2007) in Gulab Sagar
left after evaporation of the filtered sample. Total lake, Rajasthan. The water temperature fluctuated
Suspended Solids are determining as the difference between 19.03C to 36.55C in site A, 18.07C to
between the total solids and the total dissolved solids. 34.57C in site B and 18.68C to 34.89C in site C.
The pH was measured by pH meter (model no.EQ- During the present investigation, water temperature
614). Dissolved oxygen (DO) was measured by fallows more or less a similar trend as that of
modified Winklers method (1988). Free carbon atmospheric temperature, there was no great
dioxide in water was measured by titrometric method difference in water temperature. Khabade et.al.,
of APHA (1985). Alkalinity was measured by (2002) recorded the water temperature between 25C
titrometric method using phenolphthalein and methyl to 33C in Lodhe water reservoir from Tasgeon tahsil,
orange indicator. Total Hardness was measured by Sangali (M.S.). In the present investigation there is a
titrometric method using Eriochrome Black-T gradual lessening in the temperature of water from
indicator. August to December and it increases till May. Thus it
shows the temperature of water affected by seasonal
Calcium hardness and Magnesium hardness was changes in atmospheric temperature. Transparency is
measured titrometrically by using EDTA and a physical variable significant to primary production.
Eriochrome Black-T indicator. Magnesium Hardness In the present investigation, Sacchi disc transparency
was estimated by subtracting the calcium hardness was recorded at 11.00 cm to 63.99 cm in site A, 11.78
from total hardness. Chloride was measured by cm to 67.23 cm in site B and 9.23 cm to 72.33 cm in
titrometric method using potassium chromate site C. Upadhyay (2014) recorded the values of
indicator. Biochemical Oxygen Demand was estimated transparency between 6.0 cm to 92.0 cm in ponds of
by Iodometric Method of APHA (1985). Chemical Ratanpur of Bilaspur District, Chhattisgarh. Seasonally
Oxygen Demand was measured by titrometric method maximum transparency was recorded during summer

Mahajan and Pokale , 2017
Table 1: Seasonal variation of physico-chemical parameter in Mohabala lake at site-A during 2013-14
S.N. Parameters Monsoon Winter Summer Total
1 Atm. Temp. 31.23 1.59 23.66 1.64 39.26 3.85 31.38 1.05
2 Wat. Temp. 28.75 3.15 20.48 1.36 30.04 1.24 26.42 0.87
3 Trans. 13.75 2.81 30.08 5.41 50.14 9.83 31.32 2.90
4 Cond. 274.25 173.03 154.00 57.19 590.00 22.86 339.42 64.25
5 pH 7.00 0.16 7.30 0.07 7.55 0.11 7.28 0.04
6 D.O. 2.540 0.387 4.100 0.474 1.750 0.568 2.797 0.074
7 CO2 1.575 0.363 2.900 0.548 4.250 0.269 2.908 0.116
8 Alk. Total 38.50 16.77 71.25 5.12 86.00 7.07 65.25 5.10
9 Har. TO 39.063 5.049 26.192 2.454 109.018 25.395 58.091 10.258
10 Har. MG 37.344 4.414 17.682 4.308 62.219 7.354 39.082 1.412
11 Har. CA 35.750 3.700 23.750 3.700 71.000 4.899 43.500 0.565
12 CHL 3.852 0.561 4.936 0.893 12.759 3.076 7.182 1.116
13 B.O.D. 2.850 0.610 3.950 0.502 5.375 0.853 4.058 0.146
14 C.O.D. 13.800 0.447 4.450 1.001 23.000 1.281 13.750 0.346
15 PHO 1.838 0.225 0.615 0.068 3.748 0.511 2.067 0.183
16 SUL 4.835 0.411 3.295 0.219 5.418 0.721 4.516 0.206
17 NIT 0.507 0.035 0.237 0.018 0.345 0.025 0.363 0.007
18 T.S. 800.00 0.00 400.00 70.71 600.00 70.71 600.00 33.33
19 T.D.S. 500.00 70.71 200.00 70.71 300.00 70.71 333.33 0.00
20 T.S.S. 300.00 70.71 250.00 50.00 325.00 82.92 291.67 13.59
Table 2: Seasonal variation of physico-chemical parameter in Mohabala lake at site-A during 2014-15
1 Atm. Temp. 33.14 4.69 24.24 2.95 35.65 7.88 31.01 2.04
2 Wat. Temp. 28.13 4.37 21.72 1.22 28.95 5.52 26.27 1.82
3 Trans. 21.52 6.90 38.71 2.19 55.25 5.83 38.49 2.01
4 Cond. 290.50 112.29 145.00 6.20 505.50 20.67 313.67 46.97
5 pH 7.30 0.19 7.53 0.08 8.05 0.21 7.63 0.05
6 D.O. 2.980 0.966 6.300 0.778 2.300 0.644 3.860 0.132
7 CO2 2.450 0.269 3.575 0.356 6.125 1.703 4.050 0.657
8 Alk. Total 56.50 9.53 78.25 5.63 111.25 2.59 82.00 2.84
9 Har. TO 72.855 5.193 35.804 2.460 137.735 46.215 82.131 20.013
10 Har. MG 39.844 5.971 35.182 3.224 94.719 5.190 56.582 1.156
11 Har. CA 50.750 6.016 41.250 6.016 103.500 7.921 65.167 0.898
12 CHL 8.718 0.393 13.697 1.980 24.672 1.271 15.695 0.649
13 B.O.D. 6.225 0.683 9.175 1.748 12.100 1.377 9.167 0.441
14 C.O.D. 15.700 2.304 10.950 1.135 25.250 2.012 17.300 0.497
15 PHO 2.025 0.218 2.958 0.616 5.778 1.315 3.587 0.453
16 SUL 5.148 0.925 3.390 0.651 5.383 0.146 4.640 0.323
17 NIT 0.463 0.038 0.201 0.052 0.305 0.033 0.323 0.008
18 T.S. 2407.50 819.86 750.00 111.80 1825.00 739.51 1660.83 316.55
19 T.D.S. 1380.00 379.74 325.00 192.03 1075.00 601.56 926.67 167.38
20 T.S.S. 1027.50 512.12 375.00 129.90 750.00 206.16 717.50 165.17

and minimum during the monsoon. Similar minimum during the summer. Gayathri, et.al., (2013)
observations are also seen by Oomachand (1981) recorded maximum dissolve oxygen during the winter
reported minimum transparency of water in rainy and minimum during the summer in Shoolkere Lake,
seasons and maximum in winter from lower lake of Bangalore. The higher values of dissolve oxygen during
Bhopal (M.P.). Maximum transparency was recorded winter was due low atmospheric temperature and
during summer due to, settling of particles, sand slit, intensive photosynthetic activity by plants, while
clay, high light penetration and less phytoplankton minimum values during summer was due to high
productivity and minimum during the monsoon due to atmospheric temperature and high metabolic rate of
the turbid conditions of water influenced of suspended organisms and decreased oxygen holding capacity of
slit with colloidal particles. Conductivity measures the water at high temperature. The free carbon dioxide
capacity of water to conduct an electric current. In the recorded in the present study is 1.30 mg/lit. to 8.60
present investigation the values of conductivity mg/lit. in site A, 1.10 mg/lit. to 7.20 mg/lit. in site B
recorded are 103 mhos/cm to 617 mhos/cm in site and 1.5 mg/lit. to 9.30 mg/lit. in site C. Pejawar and
A, 102 mhos/cm to 657 mhos/cm in site B and 116 Gurao (2008) reported free carbon dioxide upto 5.70
mhos/cm to 636 mhos/cm in site C. Chouhan and mg/lit in Kalwa lake and upto 9.9 mg/lit in Jail lake of
Sharma (2007) recorded the values of conductivity Thane (M.S.). Seasonally the free carbon dioxide of
between 0.80 mmhhos/cm to 1.65 mmhhos/cm in the water is maximum in summer and minimum in
lake Budha, Pushkar near Ajmer, Rajasthan. Seasonally monsoon. Maximum carbon dioxide of water in
maximum conductivity of water was recorded during summer recorded by Jain and Seethapati (1996) at
summer and minimum during the winter. Similar Halalli reservoir of Vidisha district, Madhya Pradesh.
observations also seen by Chandrashekhar and The maximum free carbon dioxide in summer is due to
Kodarkar (2004) from Saroornagar lake, Hyderabad. the high rate of decomposition of organic matter in the
Maximum conductivity was recorded during summer warmer months and high rate of respiration of aquatic
due to increase in the concentration of minerals, salts, organisms, however minimum free carbon dioxide
evaporation of water and also by discharge of during monsoon may be due to a decrease in the
domestic sewage and organic matter from the nearby photosynthetic activity of aquatic organisms.
residential areas into the lake and minimum during
the winter due to nutrients uptake by plants, increase Alkalinity is a measure of buffering capacity of the
in bacterial degradation and stabilization of water. pH water. In the present investigation the total alkalinity
measures the intensity of acidity and alkalinity of ranged between 22.00 mg/lit. to 115.00 mg/lit. in site
water. In the present investigation, the pH is range A, 32.00 mg/lit. to 128.00 mg/lit. in site B and 27.00
between 6.80 to 8.30 in site A, 6.70 to 8.50 in site B mg/lit. to 109.00 mg/lit. in site C. Garg et.al., (2010)
and 6.60 to 8.40 in site C. Seasonally the maximum pH recorded the of total alkalinity values between 64.25
was recorded during summer and minimum during mg /lit. to146.25 mg/lit. in Ramsagar reservoir of
monsoon similar findings were also founded by Jakher Datia District, Madhya Pradesh. Seasonally maximum
and Rawat (2003) observed the maximum pH of water values of alkalinity founded during summer and
during summer. Maximum pH was recorded during minimum during rainy season Goldman and Wetzel,
the summer season due to high photosynthetic activity (1963) and Sreenivasan (1966) found maximum
in water. Dissolved oxygen is important for aerobic alkalinity during the summer which declined
metabolism of aquatic ecosystem. In the present subsequently in the monsoon. Total alkalinity was
investigation the values of dissolve oxygen recorded maximum during summer due to decrease in water
ranges between 1.2 mg/lit. to 7.2 mg/lit. in site A, 1.3 level and also by entry of more domestic waste,
mg/lit. to 7.5 mg/lit. in site B and 1.1 mg/lit. to 6.5 sewage attributed to increase in the rate of
mg/lit. in site C. Similar findings were also observed by decomposition from which CO2 is liberated, which
Pawar and Pulle (2005) reported the value ranges of reacts with water to form bicarbonates, due to which
dissolved oxygen from 2.8 mg/lit to 9.6 mg/lit in is total alkalinity increases in summer and minimum in
Pethwadaj dam, Nanded (M.S.) and Rawat and Jakher monsoon due to dilution caused by the rain water
(2007) reported the dissolve oxygen between 1.94 during monsoon This was further supported by many
mg/lit. to 9.25 mg/lit. in a fresh waterbody of the researchers, Moss (1973) and Wetzel (1983). Similar
desert of Rajasthan, India. Seasonally the maximum observations were also made by Bhongade and Patil
dissolved oxygen was recorded during the winter and (2010), Patil (2012) and Gayathri (2013).

Table 3: Seasonal variation of physico-chemical parameter in Mohabala lake at site-B during 2013-14
1 Atm. Temp. 31.04 1.54 23.76 1.40 39.33 3.87 31.38 1.13
2 Wat. Temp. 29.14 3.65 19.76 1.57 28.75 2.34 25.88 0.86
3 Trans. 18.76 4.50 34.07 5.04 42.93 8.97 31.92 1.99
4 Cond. 381.50 38.64 209.00 55.98 581.00 47.71 390.50 7.08
5 pH 7.03 0.19 7.40 0.12 7.98 0.13 7.47 0.03
6 D.O. 2.825 0.750 3.450 0.826 1.600 0.308 2.625 0.228
7 CO2 1.475 0.249 3.025 0.668 5.750 0.568 3.417 0.179
8 Alk. Total 42.25 10.11 54.25 7.76 94.00 11.38 63.50 1.50
9 Har. TO 43.988 8.593 29.351 2.127 82.359 10.643 51.899 3.629
10 Har. MG 31.073 7.783 15.884 3.071 53.274 6.407 33.410 1.978
11 Har. CA 33.250 9.523 21.250 1.479 53.500 6.538 36.000 3.320
12 CHL 4.168 0.697 6.568 0.693 17.005 2.385 9.247 0.797
13 B.O.D. 2.900 0.346 3.775 0.998 6.125 0.743 4.267 0.268
14 C.O.D. 21.450 1.322 8.350 3.093 27.000 2.615 18.933 0.748
15 PHO 2.368 0.386 0.700 0.084 5.003 0.762 2.690 0.278
16 SUL 6.445 0.383 4.825 0.669 8.430 0.800 6.567 0.174
17 NIT 0.645 0.036 0.310 0.054 0.367 0.074 0.441 0.015
18 T.S. 1675.00 334.48 625.00 147.90 725.00 43.30 1008.33 120.43
19 T.D.S. 900.00 122.47 225.00 43.30 375.00 43.30 500.00 37.32
20 T.S.S. 775.00 294.75 400.00 141.42 350.00 50.00 508.33 100.98
Table 4: Seasonal variation of physico-chemical parameter in Mohabala lake at site-B during 2014-15
1 Atm. Temp. 33.13 4.65 24.21 2.92 35.62 7.86 30.99 2.04
2 Wat. Temp. 28.10 4.41 22.19 2.71 30.43 4.92 26.91 0.94
3 Trans. 14.29 1.74 31.20 11.60 56.56 8.96 34.01 4.17
4 Cond. 295.25 168.71 157.50 48.71 447.25 21.19 300.00 64.05
5 pH 3.65 0.42 7.30 0.12 8.38 0.13 6.44 0.14
6 D.O. 2.550 0.206 6.825 0.698 2.400 0.255 3.925 0.221
7 CO2 2.750 0.112 3.700 0.524 6.600 0.430 4.350 0.177
8 Alk. Total 57.25 4.21 64.50 2.69 107.00 14.51 76.25 5.25
9 Har. TO 61.483 6.915 46.808 2.633 149.564 55.633 85.952 24.039
10 Har. MG 40.573 1.608 27.384 1.243 82.524 19.958 50.160 8.738
11 Har. CA 50.750 6.759 31.750 0.829 100.750 20.204 61.083 8.105
12 CHL 12.192 0.678 25.574 2.007 25.519 1.354 21.095 0.542
13 B.O.D. 6.225 0.164 10.250 1.645 14.675 1.460 10.383 0.659
14 C.O.D. 24.150 2.095 14.500 2.748 34.300 0.640 24.317 0.881
15 PHO 3.410 0.339 2.613 0.490 6.653 0.938 4.225 0.254
16 SUL 5.310 0.971 3.608 0.712 6.318 0.725 5.078 0.119
17 NIT 0.430 0.029 0.140 0.022 0.283 0.033 0.284 0.005
18 T.S. 2775.00 672.22 675.00 227.76 1150.00 455.52 1533.33 181.47
19 T.D.S. 1525.00 511.74 250.00 86.60 550.00 335.41 775.00 174.40

The hardness of water is mainly due to concentration the summer season in all the three sites of lakes.
of alkaline earth metals cation such as Ca ++, Mg++. In Pejawar et.al., (2004) reported low chloride range
the present investigation the total hardness was during rainy seasons and high in summer and
recorded in the range of 22.044 mg/lit. to 217.474 Minimum values of chloride recorded during rainy
mg/lit. in site A, 26.470 mg/lit. to 245.450 mg/lit. in seasons suggesting dilution by rain water and renewal
site B and 27.448 mg/lit. to 219.231 mg/lit. in site C. of water mass alter. It is high in summer may be due to
Sunkad and Patil (2004) observed the values between from evaporation and discharge of domestic sewage
59.8 mg/lit. to 217.4 mg/lit. in Fort lake of Belgaum, from nearby residential peoples. Similar observations
Karnataka, Andhra Pradesh. Seasonally the total have been reported by Chouhan and Sharma (2007)
hardness value was maximum during summer and and Narayana, et.al., (2008). Biochemical Oxygen
minimum in winter. Kataria et.al., (1996) also Demand has been used as a major of organic materials
observed the maximum value of total hardness in in an aquatic solutions which supports growth of
summer, moderate in monsoon and minimum in microorganisms. In the present investigation, the
winter at Kolar reservoir in Bhopal, Madhya Pradesh. biochemical oxygen demand values recorded are 2.1
High value in summer is mostly due to rising in mg/lit. to 13.6 mg/lit. in site A, 2.3 mg/lit. to 17.1
temperature due to which increase the solubility of Ca mg/lit. in site B and 3.2 mg/lit. to 22.5 mg/lit. in site C
and Mg salt and decrease in water volume similar of pond. Kamat et.al., (2006) reported the biochemical
observations also made by Manish Upadhyay (2014). oxygen demand values 1.23 mg/lit to 4.5 mg/lit in
In the present investigations the calcium hardness was Husalli tank and 4.8 mg/lit to 32.00 mg/lit in Purle
recorded in the range of 18.00 mg/lit. to 117.00 mg/lit. tank of Shimoga district, Karnataka.
in site A, 9.00 mg/lit. to 125.00 mg/lit. in site B and
17.00 mg/lit. to 108.00 mg/lit. in site C. In the present Seasonally the maximum biochemical oxygen demand
investigations the magnesium hardness was recorded value is recorded during summer and minimum during
in the range of 12.313 mg/lit. to 103.179 mg/lit. in site the monsoon season. Bhatt et.al., (1999) documented
A, 11.00 mg/lit. to 112.905 mg/lit. in site B and 16.146 that the maximum range of biochemical oxygen
mg/lit. to 67.103 mg/lit. in site C. Airsang, R.V. and H.C. demand in summer and minimum in winter and stated
Lakshman (2013) recorded the values of calcium that high values of biochemical oxygen demand may be
hardness between 36 mg/l to 240 mg/l, and for due to higher rate of organic decompositions and
magnesium hardness 11.6 mg/l to 129.3 mg/l. in gradual decline of biochemical oxygen demand from
Shetter lake of Navalgund, Dharwad District in monsoon followed by winter, which could be due to
Karnataka. decrease in temperature which in turn retards the
microbial activities. In the present investigation, the
Seasonally calcium and magnesium hardness was chemical oxygen demand values recorded are 3.4
minimum during winter and maximum during the mg/lit. to 28.4 mg/lit. in site A, 5.2 mg/lit. to 35.20
summer. Bhongade and Patil (2010) recorded the mg/lit. in site B and 8.8 mg/lit. to 36.60 mg/lit. in site
maximum value of calcium hardness was recorded in C of pond. Kamat et.al., (2006) observed the value of
May 2005 and minimum in August 2004 of Mohgavhan chemical oxygen demand fluctuated between 226.5
lake of Karanja (Lad) of District Washim (M.S.). mg/lit. in Husalli tank and 12 mg/lit. to 18 mg/lit. in
Maximum values of calcium and magnesium hardness Purle tank of Shimoga district, Karnataka. Seasonally
in summer is due to low water levels which increases maximum chemical oxygen demand was recorded
concentration and decomposition by bacteria of the during the summer and minimum during the winter.
organic matter in the lake. Chloride as anion occurs in Salve and Hiware (2006) observed the maximum
all natural waters. In the present investigation, the values of chemical oxygen demand in summer and
values of chloride recorded are as 3.110 mg/lit. to monsoon and minimum in winter in Wanparakalpa
26.583 mg/lit. in site A, 3.398 mg/lit. to 28.549 mg/lit. Reservoir, Nagapur of Dist. Beed and stated that
in site B and 5.638 mg/lit. to 35.585 mg/lit. in site C. minimum value of chemical oxygen demand in winter
Ahmed and Krishanamurthy (1990) recorded the due to fewer amounts of water, death and decay of
chloride range of 8.58 to 29.52 ppm and 13.37 to 46.64 aquatic flora and fauna and minimum chemical oxygen
ppm during 1976 to 1977 and 1977 to 1978, demand in winter due to settlement and dilution
respectively. Seasonally minimum chloride was effect. Maximum chemical oxygen demand value is
recorded during rainy season and maximum during recorded during the summer season may be due to

Table 5: Seasonal variation of physico-chemical parameter in Mohabala lake at site-C during 2013-14
1 Atm. Temp. 31.23 1.53 23.59 1.55 39.27 3.92 31.36 1.12
2 Wat. Temp. 28.69 2.41 20.56 1.67 30.80 1.79 26.69 0.32
3 Trans. 17.18 4.73 33.72 7.88 44.29 4.80 31.73 1.47
4 Cond. 415.75 86.96 340.75 97.96 580.00 44.72 445.50 22.95
5 pH 6.83 0.18 7.43 0.11 8.00 0.07 7.42 0.04
6 D.O. 2.825 1.415 3.700 0.552 1.950 1.021 2.825 0.353
7 CO2 1.600 0.071 3.375 0.753 4.775 0.589 3.250 0.291
8 Alk. Total 46.25 11.34 59.50 4.72 93.75 4.97 66.50 3.07
9 Har. TO 49.379 5.744 30.223 2.998 108.592 13.678 62.731 4.528
10 Har. MG 29.202 2.712 14.052 1.608 60.013 4.788 34.422 1.319
11 Har. CA 34.000 2.121 19.500 2.179 60.500 6.874 38.000 2.227
12 CHL 8.476 2.049 12.829 1.482 18.766 1.489 13.357 0.265
13 B.O.D. 3.475 0.363 5.250 0.844 7.200 0.707 5.308 0.202
14 C.O.D. 15.400 2.358 11.150 1.824 28.500 4.736 18.350 1.266
15 PHO 2.520 0.334 1.380 0.364 4.724 0.291 2.875 0.030
16 SUL 5.903 0.556 5.458 0.414 7.510 0.491 6.290 0.058
17 NIT 0.518 0.068 0.173 0.044 0.323 0.023 0.338 0.019
18 T.S. 3800.00 452.77 1050.00 229.13 2050.00 335.41 2300.00 91.34
19 T.D.S. 2125.00 178.54 500.00 254.95 925.00 178.54 1183.33 36.02
20 T.S.S. 1925.00 711.95 575.00 108.97 1125.00 303.11 1208.33 251.31
Table No. : 6: Seasonal variation of physico-chemical parameter in Mohabala lake at site-C during 2014-15

1 Atm. Temp. 33.15 4.65 24.26 2.96 35.61 7.87 31.00 2.04
2 Wat. Temp. 27.99 4.90 20.47 0.80 29.18 4.45 25.88 1.84
3 Trans. 17.27 2.71 35.53 6.91 59.05 12.33 37.28 3.93
4 Cond. 259.50 64.49 158.50 51.24 467.50 64.06 295.17 6.15
5 pH 7.23 0.08 7.35 0.11 8.05 0.32 7.54 0.11
6 D.O. 2.200 0.982 4.700 1.206 3.225 1.487 3.375 0.207
7 CO2 3.550 0.269 4.075 0.642 7.775 1.132 5.133 0.353
8 Alk. Total 54.50 6.65 76.50 7.83 105.50 2.69 78.83 2.20
9 Har. TO 68.765 4.731 55.493 3.412 135.168 48.763 86.476 21.075
10 Har. MG 35.702 4.386 24.052 1.608 84.513 6.349 48.089 1.945
11 Har. CA 44.000 6.285 30.500 2.062 97.500 6.344 57.333 2.005
12 CHL 15.197 1.939 20.716 3.198 29.278 4.049 21.731 0.867
13 B.O.D. 7.925 0.521 14.150 2.134 19.950 2.156 14.008 0.765
14 C.O.D. 13.000 3.842 7.850 1.090 30.150 5.833 17.000 1.945
15 PHO 4.075 0.895 2.868 0.480 6.840 0.690 4.594 0.170
16 SUL 4.148 0.143 3.398 0.255 6.120 0.689 4.555 0.235
17 NIT 0.471 0.019 0.225 0.027 0.300 0.073 0.332 0.024
18 T.S. 4250.00 502.49 1275.00 216.51 1250.00 269.26 2258.33 124.26
19 T.D.S. 2650.00 807.77 650.00 259.81 625.00 178.54 1308.33 279.45
20 T.S.S. 1475.00 601.56 525.00 326.92 625.00 238.48 875.00 154.59

higher decomposition activities and low level of water. monsoon and minimum during the winter in a
Minimum in winter is due to low temperature, low trophical lake of Jodhpur, Rajasthan. Maximum value
rate of decomposition activities. Phosphorous is one of nitrate was recorded during the monsoon due to
of the most important nutrients of the living entry of domestic sewage, surface run off. Water as a
organisms. The major sources of phosphorus are universal solvent, dissolved different type of materials
detergents, domestic sewage, agricultural effluents, as compare to other solvents (Welch, 1952). Total
and industrial waste waters, weathering of phosphate solids include all of the solid constituent of water,
bearing rocks and leaching of soil. In the present organic or inorganic or the total of the suspended and
investigation, phosphate is recorded between 0.550 dissolved solids. In the present investigation, the
mg/lit. to 7.430mg/lit. in site A, 0.560 mg/lit. to 5.990 values of total solids range between 300 mg/lit. to
mg/lit. in site B and 1.030 mg/lit. to 7.680 mg/lit. in 3600 mg/lit. in site A, 400 mg/lit. to 1900 mg/lit. in
site C. Manish Upadhyay (2014) recorded the values site B and 800 mg/lit. to 1600 mg/lit. in site C. The
of phosphate between 0.12 mg/lto 12.38 mg/l in values of total dissolved solids range between 100
ponds of Ratanpur, Bilaspur District, Chhattisgarh. mg/lit. to 1400 mg/lit. in site A, 200 mg/lit. to 1100
Seasonally maximum phosphate was recorded during mg/lit. in site B and 200 mg/lit. to 800 mg/lit. in site C.
summer and minimum during winter similar and the values of total suspended solids range
observations also made by Patil, et.al., (2008) also between 200 mg/lit. to 2200 mg/lit. in site A, 200
founded maximum phosphate during summer and mg/lit. to 900 mg/lit. in site B and 400 mg/lit. to 900
minimum during the monsoon in two water bodies in mg/lit. in site C. Narasimha et.al., (2011) recorded the
Washim district (M.S.). Maximum phosphate was values of total solid between 556.50 mg/l to 785.75
recorded during summer due to entry of domestic mg/l in a Perennial Tank in Warangal District, A.P.
sewage in pond water and due to the washing of large Ugale (2011) reported the values of total dissolved
amount of clothes by residential peoples. The sulphate solid between 210 to274 mg/l. in Jakekur project
ion occurs naturally in most water supplies and in (Reservoir). Kulkarni et.al., (2013) recorded the values
wastewater also. During the present investigation, the of total suspended solid between 50mg/lit. to 90
values of sulphate recorded are 2.310 mg/lit. to 6.300 mg/lit in Kala Talao, Kalyan. Seasonally total solid,
mg/lit. in site A, 2.460 mg/lit. to 9.120 mg/lit. in site B total dissolved solid and total suspended solid follow
and 3.110 mg/lit. to 8.210 mg/lit. in site C. Sushmitha the same trend maximum in monsoon and minimum in
(2015) recorded the average amount of sulphate in winter season. Maximum values in in monsoon due to
the tank was found to be 8.01 mg/l in Kavoor tank in siltation, precipitation, increased surface runoff from
Dakshina Kannada, Karnataka. Seasonally maximum nearby catchment area, sand, clay and other
sulphate level was recorded during summer and suspended particles entering into lake with the rain
minimum during winter. Ahmad and Krishnamurthy water.
(1990) observed maximum sulphate during summer
and minimum during the winter from Wohar
reservoir, Aurangabad (M.S.). Higher concentration of CONCLUSION:
sulphate in summer is mainly due to utilization of
sulphate by aquatic plants and discharge of domestic Lakes are important part of urban as well as rural
sewage from residential people. Minimum values of ecosystem. Lakes are comparatively small in size but it
sulphate in winter probably due to biodegradation and perform significant role in environmental, drinking
low water level (Gayathri, 2013). Nitrate content is water, recharging groundwater social and economic
most important parameter in studies of pollution. In functions. From the above observations, it may be
the present investigation, the values of nitrate range concluded that the physico-chemical characteristics of
between 0.134 mg/lit. to 0.547 mg/lit. in site A, 0.110 three sites of Mohobala lake water varied
mg/lit. to 0.660 mg/lit. in site B and 0.110 mg/lit. to consideratbly and showed characteristic change in
0.560 mg/lit. in site C. Mohekdar et al., (2003) relation to the seasonal changes.
observed the values of nitrate between 0.1 mg/lit. to
0.48 mg/lit. from Manjara reservoir (M.S.). Seasonaly
maximum nitrate was recorded during the monsoon Conflicts of interest: The authors stated that no conflicts of
interest.
and minimum during the winter. Jakher and Rawat
(2003) observed the maximum level during the

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Gowri P (2011) Seasonal changes in the ostracod
Airsang RV and HC Lakshman (2013) Diversity of
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Perennial tank in Warangal District, A.P. International
Shetter lake of Navalgund, Dharwad District in Karnataka,
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India. Science Research Reporter. 3 (2): 139-134.
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APHA (1985) Standard methods for the examination of
Narayana J, Puttaiah ET and Basavaraja D (2008) Water
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Shikaripur, Shimoga, Karnataka. J. Aqua. Biol. 23 (1): 59-
Bhatt LR, Lacoul P, Lekhak HD and Jha PK (1999) Physico- 63.
chemical characteristics of Taudaha lake, Kathmandu. Poll.
Oomachand DL (1981) Ecological studies on lower lake of
Res. 18 (4): 353-358.
Bhopal M.P. with special reference to benthic fauna. Ph.D.
Bhongade SS and Patil GP (2010) Ecological status of Thesis, Bhopal University, Bhopal.
Mohgavhan lake Karanja (lad) Dist. Washim (M.S.) with
Patil GP, Kedar GT and Yeole SM (2008) Zooplankton
reference to pisciculture. Ph.D. Thesis.S.G.B. Amravati
biodiversity study of two water bodies in Washim District
University, Amravati.
(M.S.). J. Aqua. Biol. 23 (1): 13-17.
Bhongade SS and Patil GP (2010) Limnological study of
Patil JG (2012) assessment of physico-chemical parameters
Mohgavhan Lake, Karanja (Lad) DistrictWashim, (M.S.)
of Adan Dam Reservoir, Karanja (Lad), Dist. Washim
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Chandrashekher SV, Nalini C and Kodarkar MS (2004)
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Pejavar Madhuri, Vaishali Somani and Goldin Quadros
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90-92. 2017| Published by IJLSCI

RESEARCH ARTICLE
Effect of weed green manure, compost manure and vermicompost

on productivity of Spinach
Parbhankar RL and Mogle UP
Department of Botany JES, R. G. Bagdia Arts S. B. Lakhotia Comm. and R. Bezonji Sci. College, Jalna (MS) India
E mail- rekhajain0312@gmail.com
Received: 23.06.2017 Organic manures were prepared from common weeds like Tephrosia and
Accepted: 12.08.2017 Achyranthus. Spinach seeds was sown at the seed rate 30 kg/ha, frequent
Published : 27.09.2017 irrigation was given as per requirement. In a course of time 3 regrowths
were studied after 41, 76 and 111 days of sowing. Productivity of spinach
Editor: shows maximum amount of yield produced by weed vermicompost, weed
Dr. Arvind Chavhan compost and green manure as compared to chemical fertilizers and control.
Cite this article as: Key words: Weed organic manure, Green Manure, Compost, Spinach
Parbhankar RL and Mogle UP productivity.
(2017) Effect of weed green manure,
compost manure and vermicompost
on Productivity of Spinach;
International J. of Life Sciences, 5 (3): INTRODUCTION
447-450.
Weed biomass is one of the easily available source of organic matter and
plant nutrients. Economic utilization of this weed biomass for the
production of various compost is will open a new horizon. Generally weeds
Copyright: 2017| Author (s), This like Cassia, Crotalaria, and Achyranthus are used as green manures (Chamle,
2007). The function of green manure is to add organic matter to the soil
(Kipps, 1970). Many weeds can be used for composting and
Derivs License, which permits use vermicomposting as nutrient source for many crops. (Naikwade et al.,
and distribution in any medium, 2011a, Naikwade et al., 2011b, Ghadge et al., 2013).
MATERIAL AND METHODS
The fresh green leafy vegetation of Tephrosia and Achyranthus collected
from nearby wasteland during the early hours of the day at 10-20
%flowering stage, chopped into small bits (2-3 cm) by the traditional iron
cutter. The weed plant material was incorporated into the pots at the rate of
13333 kg/ha about 5-10 cm in the soil as green manure (GM). The same
amount of weed vegetation was used for the preparation of compost (CM),
and vermicomposting (VM).
Chemical analysis
The leaf chlorophyll content (a,b and total) were estimated following Arnon
(1961), using 80% acetone as a solvent for extraction of pigments. Nitrogen
(N) content of the samples were estimated by Micro Kjeldahl method
(AOAC, 1965).

Parbhankar and Mogle, 2017
Statistical Analysis RESULTS AND DISCUSSION

All the results were statistically analyzed using
analysis of variance (ANOVA) test and treatments The fresh weight and dry weight of leaves was found
means were compared using the least significant higher in VMA in all the three harvest, but was
difference (C. D. p = 0.05) which allowed followed by different orders of treatments. During first
determination of significance applications (Mungikar, harvest it was followed by VMT, GMT, and GMA. It was
1997). found equal in treatment of COT and COA. It was
minimum in FER and control. But during second
Spinach is a short duration vegetable crop, it requires harvest order was again different, it was followed by
proper and sufficient N and K for regular growth treatments COA, GMA, VMT, COT and GMT. During
(Premshekhar and Rajshree, 2009). Pot experiment third harvest the highest treatment was followed by
was conducted to evaluate the role of weed manures VMT, GMA, COA, COT, and GMT. It was minimum in
for improving biochemical content in spinach. FER and control. (Table 1, 2, 3).
Table-1. Effect of weed manures on spinach. (Age of the plant: 41 DAS).
FW % Kg / ha
Treatments
(gm.) DW N CP FW DW N CP
GMT 71.30 7.10 1.32 8.25 11572 1152 214 1338
GMA 67.00 6.60 1.42 8.87 10874 1071 230 1439
COT 72.60 7.20 2.12 13.25 11783 1169 344 2150
COA 72.60 7.20 2.40 15.00 11783 1169 390 2434
VMT 78.60 7.80 2.56 16.00 12757 1266 415 2596
VMA 82.30 8.00 2.65 16.56 13406 1298 430 2688
FER 56.60 6.40 1.00 6.25 9168 1039 162 1014
CON 55.00 5.50 0.92 5.75 8927 893 133 933
S. E. 559 46 42 256
C.D. (0.05) 1325 109 100 607
ABBREVIATIONS:
CON = Control; CFU = Chemical fertilizer urea; GMT = Green manure Tephrosiaperpurea; GMA = Green manure
Achyranthesaspera; COT = Compost Tephrosiaperpurea; COA = Compost Achyranthesaspera; VMT = Vermicompoost
Tephrosiaperpurea; VMA = Vermicompost Achyranthesaspera. S. E. = Standard error. C. D. = Critical difference

FW % Kg / ha
Treatments
GMT 58.30 5.90 2.18 13.62 9462 958 354 2210
GMA 76.60 6.60 2.65 16.56 12432 1071 430 2687
COT 66.60 6.30 3.15 19.68 10809 1022 511 3194
COA 95.00 9.00 3.80 23.75 15419 1461 617 3855
VMT 73.30 7.70 3.90 24.37 11897 1250 633 3955
VMA 110.00 11.00 2.68 16.75 17853 1785 435 2718
FER 42.30 4.40 1.10 6.88 6865 714 179 1117
CON 39.00 4.10 0.84 5.50 6330 666 136 893
S. E. 1399 133 67 402
C.D. (0.05) 3316 315 159 953

Effect of weed green manure, compost manure and vermicompost on Productivity of Spinach
FW % Kg / ha
Treatments
GMT 68.00 6.80 3.10 19.31 11036 1103 503 3134
GMA 79.00 8.00 2.66 16.62 12822 1298 432 2697
COT 68.30 7.00 3.16 19.75 11074 1136 512 3205
COA 76.00 7.60 4.66 29.12 12355 1233 756 4726
VMT 90.00 8.70 4.20 26.25 14607 1412 682 4260
VMA 98.30 9.80 3.28 20.50 15954 1591 532 3327
FER 60.00 4.80 1.20 7.50 9738 779 195 1217
CON 47.30 4.70 0.88 5.80 7677 762 143 941
S. E. 940 102 75 467
C.D. (0.05) 2228 242 178 1107
Organic manures are source of

macro as well as micronutrient and
plant growth promoting molecules,
which together lead to good crop
yields (Mader et al., 2002).
The content of nitrogen and crude

protein was found maximum in the
treatment of VMA followed in order
by VMT, COA, COT, GMA, and GMT.
It was minimum in FER and control
during period of first harvest.
During the period of second
Fig-1.Estimation of chl-a, chl-b and total chl (age of the plant 41 days)
harvest nitrogen and crude protein
was highest in the treatment of
VMT followed in order by COA,
GMA, VMT, COT, and GMT. It was
found minimum in FER and control.
During third harvest it was highest
in treatment of COA, followed by
VMT, VMA, COT, GMA, and GMT. I t
was minimum in FER and control.
(Table 1, 2, 3).
On the basis of statistical analysis it

has been observed that all the
values of fresh weight, dry weight,
nitrogen and crude protein was
statistically significant in all the
Fig-2. Estimation of chl-a, chl-b and total chl (age of the plant 76 days) treatments. The values of GMT,
GMA, COT, COA, VMT, and VMA
were statistically significant over
the CON, while the values of FER
was non-significant over the CON.

Parbhankar and Mogle, 2017
Fig-3. Estimation of chl-a, chl-b and total chl (age of the plant 111 days)
Chlorophyll content REFERENCES

Chlorophyll a, chlorophyll b and total chlorophyll
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Mungikar AM (1997) An Introduction to Biometry,
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Achyranthus was best as compared to compost and Agronomy, Tata M.C. Graw Hill, and Publishing
Company Ltd., Bombay
green manure. Masder P, Filebach A, Dubois D, Gunst L, Fried P and Niggli U
(2002) Soil Fertility and Biodiversity in Orgnic Farming
Conflicts of interest: The authors stated that no conflicts of Science, 296:1694-1697.
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chemical fertilizer on yield, morpho-physiological and
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Eng., 2(1) I: 19-22

RESEARCH ARTICLE
Physico-Chemical parameters of Nizam sagar dam with respect to

fish production Phytoplankton, Zooplankton
Totawar DV and Tamlurkar HL
P.G. Dept. of Zoology, Yeshwant Mahavidhylay, Nanded, MS, India

Received: 12.07.2017 The main purpose of Nizam sagar dam is fish culture, irrigation, drinking,
Accepted: 13.08.2017 agriculture and also for domestic purpose. The present investigation deals
Published : 27.09.2017 with the physica-chemical parameters of Nizam sagar dam for fisheries
management and culture of fish. The study was carried out for a period of
Editor:
twelve months during the academic year 2014. The parameters such as
Dr. Arvind Chavhan
water temperature. pH, total alkalinity, dissolved oxygen and carbon
Cite this article as: dioxide were estimated. The pH shows alkaline trend in Nizam sagar dam.
Totawar DV and Tamlurkar HL The water of Nizam sagar dam is found to be more suitable for fish culture.
(2017) Physico-Chemical parameters
of Nizam sagar dam with respect to Key words: Reservior, Physico-chemical parameters, and fish culture.
fish production Phytoplankton,
Zooplankton; International J. of Life
Sciences, 5 (3): 451-454.
INTRODUCTION
Acknowledgements
Authors are thankful to the Head,
Department of Botany for providing The fresh water bodies of India includes a large number of rivers, ponds,
facilities and the financial assistance dams, impoundments and lakes. The riverine system, have ideally the
of UGC is gratefully acknowledged. quality of water should be assessed on the basis of physic-chemical and
biological parameters on order to provide the complete spectrum of
Copyright: 2017| Author (s), This information of the purpose fisheries management. Nizam sagar dam is
is an open access article under the located near Nizam sagar village at the border of Maharashtra of Nizamabad
terms of the Creative Commons district. Dam is constructed on Manjira. a major source for irrigation and
fish production and also beneficial for drinking, agriculture purpose. It is a
deep reservoir with lot of submerged weeds and various species of fishes.
provided the original work is The catchment area of the reservoir is about 312-84 sq. kms.
adaptations are made. MATERIALS AND METHODS
For the purpose of study, the water samples were collected from the
reservoir for a period of twelve months during the year 2014 from January
to December. Four sampling spots namely A. B. C and D are selected for
study. The temperatures were recoreded at the time of sampling, on the
spot using centigrade thermometer. pH was measured with standard pH-
meter. Other parameters were estimated by the procedures given in
Trivedy and Goel (1984), also from methodology by Kodarkar.

Totawar and Tamlurkar, 2017
RESULTS AND DISCUSSIONS Temperature

It plays an important role in thermal stratification
The physic-chemical parameters for 12 months which have some effect on chemical and biological
recorded are given in the Table-1 The river water, activities of aquatic media like dissolved oxygen,
which is lotic, becomes lentic in the reservoir. Hence carbon-di-oxide, water and air temperature go more
the reverine fishery have different ecology compared or less hand in hand. The water temperature
to reservoir fishery. Thus to study the fish culture in a recoreded at morning hour ranges between 17 to 39.3
reservoir is important from the point of the ecology of 0c at all the four station. Maximum temperature is
the reservoir as said by Shrivastava (2000). The recorded in the months of May and June 2014.
climatic factors such as rainfall, temperature,
atmospheric pressure and humidity help in Hyderogen-concentration
understanding the complex process of interaction pH is the scale of intensity of acidity and alkalinity of
between the climate and the biological process in water and measures the concentration of H+ ions. Most
water bodies. Impinging. of the biological processes and biological reactions are
pH dependent. Swingle (1967) stated that waters
Solar radiation and atmospheric temperature bring having a pH range of 6.5 to 9.0 as recorded before day
about interesting changes in aquatic ecosystem, which break are most suitable for pond culture and those
play significant role in the fish culture. Many workers having pH values of more than 9.5 (alkanline) as
as, Pandey and Tripathi (1988), Pulle (2000); Pawar unsuitable, because in the later, CO3 is not available.
(2002), Wagh (1998) studied the role of climatic Fish dies at pH 11. Acidic waters reduce the appetite
factors in understanding the ecology of aquatic of the fish. Their growth and tolerance to toxic
ecosystem. substances. Acidic water, carries toxicity of H2S4
copper and other heavy metals.
Table 1. Monthly variations of physic-chemical parameters of Nizam sagar dam in 2014

Parameters Spots Months
Jan. Feb. Mar. April May June July Aug. Sept Oct. Nov. Dec.
Water A 21.60 25.50 26.00 30.10 39.20 38.00 31.00 28.00 22.00 21.00 19.00 18.00
Temerature
(0C) B 22.00 26.10 27.20 31.00 38.50 39.10 31.00 28.00 22.50 20.40 19.40 18.20
C 22.50 27.00 27.50 31.50 39.00 38.50 31.00 28.50 22.00 20.20 19.60 18.60
D 22.50 27.50 28.00 31.60 39.30 38.40 31.50 29.00 22.50 20.00 19.80 18.80
pH of Water A 7.63 7.55 7.30 7.35 7.32 7.34 7.40 7.50 7.56 7.58 7.71 7.85
B 7.64 7.56 7.32 7.35 7.32 7.35 7.40 7.52 7.55 7.58 7.70 7.86
C 7.64 7.57 7.32 7.36 7.30 7.34 7.42 7.52 7.58 7.57 7.72 7.88
D 7.63 7.56 7.30 7.38 7.30 7.34 7.42 7.42 7.50 7.57 7.70 7.87
Total A 134.0 130.8 129.9 131.0 132.2 132.8 136.2 139.9 138.1 137.9 128.0 131.3
alkalinity
B 134.0 130.8 129.9 131.3 132.1 132.9 136.2 139.8 138.0 137.9 128.1 131.2
(mg/1)
C 134.2 130.7 129.8 131.5 132.1 132.7 136.4 140.0 138.1 137.8 128.1 131.5
D 134.4 130.6 129.8 131.6 132.1 132.6 136.4 140.0 138.0 137.9 128.0 131.4
Dissolved A 9.50 9.10 6.00 6.40 6.10 7.20 8.50 9.80 8.90 8.86 8.90 9.33
Oxygen B 9.41 9.00 5.80 6.50 6.30 7.20 8.57 9.86 8.85 8.87 8.91 9.30
(mg/1) C 9.50 9.10 6.10 6.80 6.60 7.00 8.54 9.79 8.89 8.88 8.89 9.25
D 9.61 9.10 6.10 6.60 6.20 7.30 8.50 9.87 9.01 8.87 8.90 9.22
Carbon A 4.41 3.10 4.25 3.11 3.96 4.42 6.10 6.39 5.91 4.98 3.99 3.82
Dioxide B 4.60 3.12 4.32 3.10 3.97 4.45 6.99 6.30 5.99 4.11 3.10 3.10
(mg/1)
C 4.55 3.11 4.42 4.10 3.99 4.40 6.99 6.32 5.91 4.11 3.10 3.10
D 4.48 3.11 4.35 4.41 3.10 4.32 6.10 6.41 5.99 4.99 3.10 3.10

Physico-Chemical parameters of Nizam sagar dam with respect to fish production
The fish gets prone to attacks of parasites and diseases organisms. As CO2 is highly soluble in water, it is found
in acidic water. hence pH is considered as an indicator to be in larger amount in polluted water compared to
of overall productivity that causes habitat diversity fresh water bodies. CO2 has a great effect on on
(Minns, 1989). The pH recorded, ranged from 7.30 to photosynthesis which effect again onfish growth.
7.88 at all the four stations Dwivedi and Pandey (2002) found the free carbon
dioxide high in pre-monsoon and monsoon period and
Total alkalinity low in winter. The values of CO2 ranges from 3.81 to
The water that can neutralize the acid is called alkaline 6.81 mg/1 at all four stations. High concentrations of
water. Acidic water is danger for fish growth, the free carbon dioxide more than 20 ppm is toxic to fish.
reservoir or ponds should have alkalinity so that the
acids can be neutralized and fish production is
possible. The alkalinity might be due to the high pH or CONCLUSION
it may be caused by cations of Ca, Mg, na, k, NH and Fe
combined either as CO3 or bicarbonate as hydroxides. The water of Nizam sagar dam is found to be more
At pH values less than 8.3 but more than 4.5 practically suitable for fish culture. The water is productive
no CO2 is present, but free CO2 and CO2 and having the maximum Alkalinity 139 mg/1 also less
bicarbonate may be present, (Jhingran, 1991). He also than 8 pH, DO and CO2 of the water is measured within
concluded that many fish ponds and rivers have the the renge of 5 to 10 mg/1 which is th suitable
total alkalinity values equivalent to 10 to 50 ppm condition for fish growth. Hence Nizam sagar dam
CaCO3. According to Alikunhi (1967) in the highly having the large area for catchment can be utilized for
productive water alkalinity reach to over 100 ppm and the production of fish on a large scale and a variety of
according to Schaperlaus (1933) most productive species can be cultured.
water is that which titrates 200 to 500 ppm equivalent
CaCO3. The total alkalinity values at Nizam sagar dam
were found to be in the range of 128 mg/1 to 138 Conflicts of interest: The authors stated that no conflicts of
mg/1. interest.
Dissolved oxygen
Do is one of the important parameter in water quality REFERENCES
assessment. Its presence is essential to maintain the
Alikunchi (1957) Fish culture in India. F.M. Bull. Indian
higher form of biological life in the water. The
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wastewater is determined largely by the oxygen
APHA (1985) Standard Methods for the examination of
balance in the system. water and waste water. 16th edition. American Public
Health Association, New York. PP : 1193.
Oxygen is formed by absorption from the atmosphere Diwedi BK and GC Pandy (2002) Physicochemical factors
at the surface of pond and by photosynthesis of the and algae diversity of two ponds (Girija Kund and
chlorophyll bearing organisms inhibiting in water Maqbara pond). Faizabad, India. Poll. Res. 21 (3) : 361-
body. Thus oxygen act as an indictor of planktonic 370.
development which has a significant role in growth of Jayaraju PB, Prasadrao GDV and Sharma SV (1994) Seasonal
variation in physico-chemical parameters and diversity
fish (Jayaraju et al. 1994), the high temperature and
in Flora and Fauna of the river Manneru, a tributary to
low dissolved oxygen during summer create
river Krishna, Andhra Pradesh, India J. Aqua Biol. 9 (1 &
favourable condition for the development of blue 2) 19-22
green algae. The salinity and oxygen are inversely Kodrakar MS (Ed) (2006) Methodology for water Analysis
proportion and therefore low oxygen results in high Publ. 2 IAAB, Hyderabad. 3rd edition PP: 106.
salinity, which affect the fish production. The dissolved Minns CK (1989) Factors affecting fish species richness in
oxygen recorded at four station of Nizam sagar dam, Ontario lakes. Transaction of the American Fisheries
ranges between 5.60 to 9.80 mg/1. Society, 118 : 533-545.
Pandey SN and Tripathi AK (1988) Schizemeris Lubleoni
(Kuetz) An indicator of organic pollutin, Geobios.15
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Pawar SK (2000) Hydrobiology of Sirur Dam near Mukhed in
is produced as a result of respiration of aquatic Nanded District Ph.D. Thesis, SRTM, Universtiy, Nanded.

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Pulle JS (2000) Bio-monitoring of Isapur Dam water Ph.D.

Thesis, SRTM University, Nanded.
Srivastava SC (2000) A text book of Fishery Science and
Indian Fisheries Kitab Mahal Publishers.
Singhal RN, Swarnajeet and Devis RW (1986) The Physico-
chemcial Environment and the Plankton of Managed
Ponds in Haryana. India Proc. Indian Acad. Sci. (Sec B).
95 (3) : 253-263
Scharperclause W (1933) Text book of Pond culture :
Rearing and keeping of Carp. Trout and allied fishes (Tr.
Fhund). Fish Leaflet. Washington, (311) : 260 P.
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Publishing Corporation, (India) Delhi PP : 280.
.

RESEARCH ARTICLE
A new species of cestode Lytocestus paithanensis (Lytocestus

Cohn, 1908) from Clarias batrachus at Paithan, MS, India
Kale SS
Department of Zoology, Shri Kumarswami Mahavidyalaya, Ausa,, Dist. Latur M.S. 413520, India
Email: Sanjaykale.sks@gmail.com

Received: 21.07.2017 The present communication deals with description of new species
Accepted: 13.08.2017 Lytocestus paithanesis n.sp. is reported from Clarias batrachus at paithan
Published : 27.09.2017 Dist. Aurangabad. It difference from all known species of the genus, with the
characters like, head is spatulate, narrower than body, broad base and
Editor:
narrow apex; neck medium, wide narrow anteriorly and posterioly wide;
Dr. Arvind Chavhan
gonads located near the posterior end of the worm, testes 220-250 in
Cite this article as: number, oval, preovarian, scattered in middle of worm; ovary medium H-
Kale SS (2017) A new species of shaped; large ootype ; utrus tubular and wide.
cestode Lytocestus paithanensis
(Lytocestus Cohn, 1908) from Key words: Tape worm, Lytocestus paithanensisn.sp., Clarias batrachus.
Clarias Batrachus at Paithan, MS,
India; International J. of Life
Sciences, 5 (3): 455-458.
INTRODUCTION
is an open access article under the The genus Lytocestus was erected by Cohn,1908 with its type species L.
terms of the Creative Commons adhaerens found in Clarias fuscus in Hong-Kong. This genus was first
Attribution-Non-Commercial - No confirmed by woodland (1926) who included four more species in addition
to the type species. They are L. filiformis woodland (1923) in
Murmurscasehive Egyptiansudan L. Clamersius woodland (1924) L.
properly cited, the use is non- Cunningtoni fuhrmann and bear (1925) and L. indicus Moghe (1925) (Syn.
commercial and no modifications or Caryophyllaeus indicus) from Clarias batrachus in India. The same were
adaptations are made. recorded by Mehra (1930) from Clarias magur and Ramadevi (1973) from
Clarias batrachusin India. Hunter (1927) placed the genus in sub-family of
his own namely Lytocestinae and retained only three species i.e. L.
Adhaerens, L. filiformis and L. indicus. He put the species L. chalmersius in
the genus Monobothriodides.
Subsequent workers Gupta (1961), Murhar (1963) have adhered to these

changes. Wardle and Mcleod (1952) followed Hunters classification, but
raised the status of Lytocestinae from sub-family to a family. Wardle,
Mcleod and Radinovsky (1974) suggested a new system of classification of
cestodes, who used the term cotyloda as a class and the order
caryophillidea is kept in this class. Mackiewicz (1972) included the species
L. iavanicus Bovine, 1926 Furtado, 1963 Lynsdale, 1956 and L. parvulus
Furtado, 1963 in this genus. Johri 1959 considered L.alestesi Lynsdale, 1956
concluded that it should be considered a synonym of L. filiformis woodland

Kale SS, 2017
(1923), Ramadevi (1973) described L. longicollisfrom short, thin, slightly curved, extends anteriorly and
Clarias batrachus in India. Later on Shinde and Phad measures 0.396 in length and 0.35 is breadth.
(1988) added L.marathwadaensis from Clarias
batrachus in India. All the nine species come from east The ovary in medium in size, H-shaped in appearance,
Asian countries. bilobed, lobes with irregular margin, due to the
presence of numerous short, blunt, round acini,
antero-posteriorly elongated, extends laterally upto
MATERIALS AND METHODS the corticular region of worm and measure 0.410 in
leangth and 1.028 in breadth. The lobes are connected
The present species were collected from the intestine by a wide, short, curved isthmus. The isthmus is short,
of fish clarias batrachus from Nathsagar dam, paithan, a wide tube, which measure 0.482 in length and 0.125
Dist. Aurangabad in the month of September 2007. The in breadth. The vagina is a thin tube, starts from the
cestod were fixed in 4 % formalin and stained with genital pore, runs medially and posteriorly, slightly
Harris haematoxylin. The drawing are made with aid curved, extends posterior to the isthmus, reaches and
of camera lucida. All measurement in mm. opens into the ootype and measure 1.142 in length
and0.053 in width. The ootype is large in size, oval in
DESCRIPTION: shape, situated behind the isthmus of the ovary,
slightly obliquely placed and measure 0.417 in length
Nine specimens, of the cestode parasites, were and 0.267 to 0.357 in breadth.
collected from the intestine of a fresh water fish,
clarias batrachus at paithan Dist. Aurangabad, M.S. The genital pores are large in size, oval in shape,
India in the month of Septeber,2007. The parasites are anterior to the uterine coils, in the middle region of the
stained with Harris haematolixin and prepared whole worm, opens in the middle of the same and measure
mount slides, for anatomical studies. 0.071 in length and 0.035 to 0.053 in breadth.
The head is spatulate, marked by narrower than the The vitelline follicles are small in size, oval in shape, in
body, broad at the base, narrow at the apex, with blunt one to two rows, on each lateral side, lateral to the
tip and measure 1.250 in length and 0.714 to 0.357 in testes, in corticular region and measure0.035 to 0.089
breadth. in length and 0.035 to 0.053 in width.
The neck is medium in length, wide, narrow anteriorly, The uterus is tubular, a wide, transverse, convoluted
wide posteriorly, with irregular lateral margin, curved, tube, coiled, loope-shaped, pre-ovarian, starts from the
longer than broad and measure 3.124 in long and ootype narrow proximally, wider distally, crosses and
1.963 to 0.982 in breadth. extends anterior to the isthmus, coils arranged in the
pre-ovarian region, opens separately by an uterine
The gonads are situated in the posterior region and pore and measure 5.801 in length and 0.053 to 0.089
near the posterior end of the worm. The testes are in breadth. The uterine pore is large in size, oval in
numerous, small and large in size, oval in shape, 220- shape, double walled, preovarian and measure 0.353
250 in number, preovarian, scattered in middle region in length and 0.535 to 0.514 in breadth.
of the worm, unevenly distributed, not distributed in
the posterior region of the worm,in 4-5 rows ,from the
base of neck to anterior region of the cirrus pouch and RESULTS AND DISCUSSIONS
measure 0.0714 to 0.178 in length and 0.0892 to 0.160
in breadth. The cirrus pouch is medium size, oval in The Genus Lytocestus was established by Cohn in
shape, antero posteriorly placed, preovarian, opens 1908, as L. adhaerens from Clarias fuscus at Hong-
posteriorly, in the middle of the worm, curved and Kong. Later on the following species are added to this
measure 0.982 in length and 0.320 to 0.178in breadth. genus.
The cirrus is a thin tube, slightly curved, colied, 1. L. filiformis wooland, 1923 in Morlmruscaschive,
contained within the cirrus pouch and measures 0.839 Egytian Sudan.
in length and 0.053 is breadth. The vas-deferens is 2. L. indicus Moghe, 1925 in clarias betrachus Nagpur,
India.

A new species of cestode Lytocestus paithanensis (Lytocestus Cohn, 1908) from Clarias Batrachus
3. L. Altestesi Lynsdale, 1956 in Alestesinurse Sudan. 7. L. logicollis remade, 1973 in Clarias batrachus,
4. L. birmaicus Lynsdale, 1956 in clarias batrachus, India.
Burma. 8. L. marathwadaensis shinde and phad, 1988 in
5. L. Javanicus (Bovian, 1926) Mackiewics, 1972. clarias batrachus, India.
6. L. parvulus (Furtado1963) Mackiewicz, 1972.
A. Anterior region B. Middle region C. Posterior region
The worm under discussion is having head spatulate, convoluted, loop-shaped and vitellaria follicular, small
narrow at apex, broad at base; nake medium, narrow on each lateral side, in 1-2 rows.
anteriorly, wide posteriorly, testes 220-250, large, pre- After goning through the literature, the worm under
ovarian, in the central medulla; cirrus pouch medium, discussion in having testes 220-250 in number
oval, placed antero-posteriorly, in middle of the worm; (numerous) come closer to L. filiformis, L. indicus and L.
cirrus thin, slightly curved, vas- deference short, thin; longicollis.
ovary medium, H shaped in appearance, with
numerous short, round acini, in cortical region of the The present cestode, differs from L. filliformis which is
worm, with short isthmus; vagina a thin tube, runs having testes numerous round, in the central medulla;
medially; ootype large, oval; seminal receptaculum cirrus pouch small, bent between the oval concavity of
absent, genital pore large,oval; seminal receptadulum ovarian lobes,ovarysmall,each lobe with 6-11 spearate,
absent, genital pore large, oval, anterior to uterine coil, large follicles, with big spindle shaped isthmus; vagina
in middle reason of the worm; uterus tubular, wide, short, wide, a thick tube; uterus with small and large
convolutions, open by uterine pore; vitellaria

Kale SS, 2017
numerous, large, rounded, all around the testicular from Clarius batrachus. Uttar Pradesh J.
zone, in medullary parenchyma. Zool.28(3):3654-369
The present tape-worm, difference from L. indicus wich Kadam MN, Hiware CJ and Jadhav BV (1998) on a new
Caryophyllid cestode of genus Lytocestus Cohn, 1908
is having body length 27.0-40.0 and width 0.3-0.5, from Clarias batrachus. Dr. B. A. M. Uni. Aurangabad J. of
testes 230-270, round, 0.17-0.19 in dimeter extending Sci. 29(6) 143-148.
upto cirrus sac and some extand lateral to cirrus sac; Kalse AT and Shinde GB (1999) Lytocestus chalisgaonesisn. sp.
vas- deferens followed by a ductusejaculatorius, ovary (Cestoda: Caryphyllidea) from the catfish Clarias
batrachus at Chalisgaon, M.S. India Riv. Parsit. 16
with numerous follicles,connected with pipe shaped
(60):1:39-42
isthmus; ootype round, posterior to ovarian follicles;
Kaul SS, Kalse AT and Suryawanshi RB (2010) Lytocestus
wall of the uteres thick and coiled and vitellaria pre- murharisp. Nov. (Cestoda: caryophyllidea) from the
ovarian, cortular, rounded and 0.13-0.19 in dimeater. catfish Clarias batrachus (L) at Chaalisgaon.decc.curr.
Sci.3(1),73-814
The present cestode, difference from L. longicollis Khadap RM, Jadhav BV and Suryavanshi NV (2004) A new
Species of the genus Lytocestus (cohn, 1908), from
which is having 10.8-20.0 length, 0.50-0.84 width,neck claries batrachus at Aurangabad. Nat. J. of Life Sciences,
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Caryophyllidean review from catfishes of Maharashtra
(India). Flora and Fauna, 14(1):03-22.
Jawalikar JD, Pawer SB and Shinde GB (2008) A new species
Lycestus subhapradhi n. sp. (Eucestoda: Lytocestidae)

RESEARCH ARTICLE
Impact of temperature variations on RNA content of Freshwater

fish Oreochromis mossambicus
Chavan PN1 and Mali RP2
1Department of Zoology, S.G.B. College, Purna, Dist. Parbhani (MS) India

2P.G. Department of Zoology, Yeshwant Mahavidyalaya, Nanded (MS), India.
Received: 04.08.2017 The present work deals with the effect of temperature on RNA content of
Revised : 28.08.2017 Fresh water fish Oreochromis mossambicus. The fishes were exposed to
Accepted: 18.09.2017 different temperature such as 20C, 24C, 28C, 32C, 36C & 40C for 24,
Published : 30.09.2017 48, 72 and 96 hours exposures respectively. The RNA Content were
decreased at low temperature and at high temperature as compared to
Editor: control set. The values of RNA content were expressed in mg/gm wet wt. of
Dr. Arvind Chavhan tissue and plotted graphically.

Key words Temperature, RNA, Oreochromis mossambicus.
Chavan PN and Mali RP (2017)
Impact of temperature variations on
RNA content of Freshwater fish
Oreochromis mossambicus;
International J. of Life Sciences, 5 (3): INTRODUCTION
459-461.
Aquatic biota is affected by the temperature of the surrounding water
which influences the biochemical and physiological activities including food
intake, growth rate, body temperature and other body functions.
Temperature is an important factor which affects the chemical
Attribution-Non-Commercial - No compositions of various tissues of aquatic organisms (Prosser & Brown
Derivs License, which permits use 1961). Several variations in biochemical constituents of tissues have been
and distribution in any medium, associated with differences in environmental temperatures. In the natural
provided the original work is process of temperature acclimation, most of the poikilotherms adjust their
properly cited, the use is non- metabolic rates to maintain physiological activity at constant level, thus
commercial and no modifications or these animals attain a measure of independence of temperature (Prosser
adaptations are made. and Brown 1961).
Nucleic acids are the nitrogen containing compounds of higher molecular

complexes associated with protein in cell. The alterations in the DNA
content reflect the changes in RNA and protein synthesis, lead to variation
in protein profile (Raj & Selvarajan 1992). The biochemical parameters i.e.
nucleic acid plays a significant role in physiological mechanism. Hence the
aim of present study was to examine the effect of temperature on RNA
content of fresh water fish Oreochromis mossambicus.

Chavan and Mali, 2017
MATERIAL AND METHOD and minimum at 20C & 40C i.e. 2.11 and 2.12 mg/gm
wet wt of tissue respectively. Nucleic acid plays the
The fresh water fish Oreochromis mossambicus were key role in protein synthesis, it is considered as an
collected from Godavari River, Nanded (MS) with the index capacity of an organism for protein synthesis
help of local fisherman for the present investigation. (Mali and Kadam, 2010).
They were acclimated for 07 days prior to
experimentation. The varying sizes of fishes ranging In the present investigation the RNA Content at low
from 50-55 gm were selected for experiment. The temperature i.e. at 20C and high temperature i.e. at
tissue like liver and muscle were selected for present 40C were found to be decreased as compared to
study. The estimation of RNA content was determined control set. Similar observations also reported in
by the method of Orcinol (Bial, 1962). Oncohynchus mykiss Currie et. al., (2000). The decrease
in RNA content in the tissue of fish observed at high
temperature as compared to control set. The
RESULT AND DISCUSSION concentration in a cell is related to metabolic function
of tissue (Bulow et. al., 1978). The fall in RNA content
In the present investigation, the fresh water fish can be attributed to the reduced rate of protein
Oreochromis mossambicus were exposed to variable synthesis due to temperature stress. The present study
temperature up to 96 hours. The significant changes in showed that maximum amount of RNA content
RNA content were recorded in table 01 and 02. The observed at 32C & 36C as compared to control set.
results were graphically represented by bar charts. Jayprakash and Sambhu (1996) suggested that
increased RNA content in the hepatopancreas and
The present investigation showed that the maximum muscle of Penaeus indicus correlated with enhanced
amount of RNA content observed at 32C and 36C i.e. protein synthesis and growth. The increased rate of
2.50 and 2.52 mg/gm wet wt. of tissue respectively protein synthesis directly correlates with the available
Table 1: Effect of temperature on RNA content in Liver of Freshwater fish, Oreochromis mossambicus at 20C,
24C, 32C, 36C and 40C.
Sr. Exposure Exposure of Animals
No. Hours Temperature variations in C
20C 24C 281C 32C 36C 40C
(control)
1 24 2.390.031 2.270.025 2.340.016 2.530.024 2.550.033 2.370.029
2 48 2.360.016 2.250.019 2.330.017 2.530.024 2.540.033 2.350.033
3 72 2.340.016 2.280.022 2.320.020 2.520.017 2.520.017 2.320.020
4 96 2.320.020 2.220.036 2.320.020 2.500.024 2.520.017 2.280.020
(Each Value is mean of six observations SD)
Figure 1: Effect of temperature on RNA content in Liver of Freshwater fish, Oreochromis mossambicus at
20C, 24C, 32C, 36C and 40C.

Impact of temperature variations on RNA content of Freshwater fish Oreochromis mossambicus.
Table 2: Effect of temperature on RNA content in Muscle of Freshwater fish, Oreochromis mossambicus at 20C,
24C, 32C, 36C and 40C.
Sr. Exposure Exposure of Animals
No. Hours Temperature variations in C
20C 24C 281C 32C 36C 40C
(control)
1 24 2.160.035 2.190.021 2.200.024 2.240.035 2.480.036 2.200.027
2 48 2.130.025 2.170.016 2.180.028 2.250.032 2.470.032 2.170.036
3 72 2.140.024 2.160.031 2.170.018 2.230.025 2.450.036 2.150.028
4 96 2.110.020 2.140.030 2.160.017 2.210.028 2.420.018 2.120.026
(Each Value is mean of six observations SD)
Figure-2: Effect of temperature on RNA content in Muscle of Freshwater fish, Oreochromis mossambicus at
20C, 24C, 32C, 36C and 40C.
synthesis machinery (Pace et. al., 2010). The fall and Jayaprakas V and Sambhu C (1996) Growth response of
rise of nucleic acid content may be due to recovery of white prawn, Penaeus indicus to dietary L. Carnitine, J.
Asian Fisheries Science 9: 209-219
metabolic and physiological processes to attain the
Mali RP and Kadam MS (2010) Comparative fluctuation in
normal stage. Similar results were observed in fresh RNA contents of Barytelphusa guirini after exposure to
water crab Barytelphusa guirini (Mali and Chavan zinc and cadmium sulphate toxicity, Journal of Eco-
2015). Physiology Occupational Health 10: 173-176
Mali RP and Chavan PN (2015) Temperature dependent
Conflicts of interest: The authors stated that no conflicts of alterations of Nucleic acid content in various tissues of
fresh water crab Barytelphusa guirini from Nanded
interest.
Region (MS).World Journal of Pharmaceutical Research,
4:, 1396-1400.
Pace DA, Maxon R and Manahan DT (2010) Ribosomal
REFERENCES analysis of rapid rates of protein synthesis in the
Antarctica sea urchin Sterechinus neumayeri, Journal of
Biol, Bull., 218: 48-60
Bial M (1962) Orcinol reaction for ribose. Deutsche Med.
Wochschr., 28: 253 Prosser CL and Brown SA (1961) comparative animal
Physiology 2nd Edition W. B. Saunder Co. Philadelphia.
Bulow FJ, Lourn CB and Cobb CS (1978) Comparison of two
blue gill populations by means of the RNA/DNA ratio Raj DS and Selvarajan (1992) Influence of Quinalphos a
and liver somatic index. Trans. Am.Fish. Soc., 107: 799- organophoporous pesticide, on the biochemical
803 constituents of the tissues of fish Oreochromis
mossambicus, Journal of Environmental Biology, 13: 181-
Currie S, Moyes CD and Tufts BL (2000) The effects of heat
185.
shock and acclimation temperature on HSP 70 and HSP
30 m RNA expression in rainbow trout in vivo and in
vitro comparision. J. Fish. Biol. 56 ; 398-408 2017| Published by IJLSCI

RESEARCH ARTICLE
Systematic study of Cestode parasite Moniezia (Blanchariezia)

Caprae n. sp. of Capra hircus from Kasgi Tq. Omerga
Dist. Osmanabad, (MS) India
Kale Sanjay S
Department of Zoology, Shri Kumarswami Mahavidyalaya, Ausa, Dist. Latur M.S. India.
Email: sanjaykale.sks@gmail.com

Received: 12.07.2017 The new species of the genus moniezia, Blanchard 1891 collected from
Accepted: 13.09.2017 intestine of Capra hircus at kasgi Tq. Omerga Dist. Osmanabad, M.S india. It
Published : 30.09.2017 differs from all the know species of genus with charactes, scolex small,
triangular, auteriorly broad, posteriorly narrow, four suckers towards
Editor:
anterior, oval, long neck, matureproglotttids broader than long, double set
Dr. Arvind Chavhan
of reproductive organs testes 40 to 50 oval, cirrus thin, coiled, protruded
Cite this article as: within pouch, cirrus pouch near anterior margin of segment, medium
Kale Sanjay S (2017) Systematic cylindrical, curved, thin long vas deferens, ovary bilobed, medium, middle in
study of Cestode parasite the segment, vagina thin tube, posterior to cirrus pouch, ootype small, oval,
Moniezia (Blanchariezia) Caprae genital pore bilateral, small and oval.
n. sp. of Capra hircus from Kasgi
Tq. Omerga Dist. Osmanabad, Key words: Tape worm, Moniezia caprae n. sp, and Capra hircus.
(MS) India; International J. of Life
Sciences, 5 (3): 462-464.
INTRODUCTION
The genus Moniezia was eatablished by Blanchard 1891. Skrjabin and
Attribution-Non-Commercial - No Schulz (1937) divided this genus into three sub-genera as follows:
Derivs License, which permits use 1. Tnterproglottid glands groped in rosettes Moniezia
and distribution in any medium, 2. Inter proglottid glands arranged lineally (sometimes absent)
provided the original work is .....Blanchariezia
properly cited, the use is non- 3. Tnterproglottid glands absent ....Bariezia
commercial and no modifications or The present worm agrees in all the characters with subgenus Blanchariezia
adaptations are made. Skrjabin and Schulz (1937) having two species as M.(B.) Bendeni
(Moniez1879), Skarj. et Schulz 1937 and M. (B.) Pallida Moning, 1926. Later
on two more two new species were added by Shinde et al., from the host
Ovis bharal at Aurangabad, Maharashtra state, India. The present
communication deals with the description of a species Moniezia (B.) caprae
n. sp. Collected from goat Capra hircus at Kasgi, Tq. Omerga, Dist.
Osmanabad, Maharashtra state, India.
DESCRIPTION
Numerous specimens, of the cestode parasites, were collected from the
intestine of a goat, Capra hircus, at kasgi, Dist. Osmanabad, Maharashtra
state, India.

Systematic study of Cestode parasite Moniezia (Blanchariezia) Caprae n. sp. of Capra hircus
The ovary is distinctly bilobed, medium in size,

obliquely placed, situated either in the middle or
slightly posterior to the middle of the segments,
almost at 1/4th from the lateral margin of the same,
lobes nearly equal in size, with irregular margins and
measures 0.928 to 0.839 in length and 0.267 to 0.089
in width.
The vigina on each side is a thin tube, posterior to the

cirrus pouch, runs obliquely and transversely for a
long distance, enlarges and from the receptaculum
seminis, then become narrow, reaches and opens in to
the ootype and measure 1.071 in length and 0.035 in
width. The receptaculum seminis is medium in size,
oval in shape, obliquely placed and measure 0.624 to
0.570 in length and 0.213 to 0.142 in width. The
ootype is small in size, oval in shape, ventral to the
Fig . 1 isthmus and measure 0.0535 in length and 0.0714 in
width. The genital atrium is large in size, oval in shape,
The scolex is small in size, distinctly marked off from marginal, situated in the anterior half of the segments
the strobillia, simple, almost triangular, broad anterior, and measures 0.267 in length and 0.340 in width. The
narrow posterior and measures 1.022 to 0.902 in genital pores are bilateral, small in size, oval in shape,
length and 1.249 to 0.510 in width. It bears four marginal, in the anterior half of the segments and
suckers, which are medium in size, oval in shape, measures 0.035 in length and 0.089 in width.
placed in a row, towards anterior end of it and
measure 0.270to 0.247 in length and 0.303 to 0.281 in The excretory canals are thin and measures 0.089 in
width. The rostellum is absent. The neck is long, width. The vitelline gland is medium in size, oval in
almost of the same width throughout and measure shape, postovarian, compact, obliquely placed and
4.848 to 4.486 in length and 0.476 to 0.341 in width. measure 0.195 in length and 0.107 in width. They are
absent near the lateral margins, in each segment, on
The mature proglottids are broader than long, almost each side. The inter proglottidal glands are present in
four time broader than long, with concave and convex the intestinal region, which are 12 to 13 in number,
lateral margins, with double set of reproductive oval in shape, arranged in pairs and measures 0.142 to
organs, having projections at the posterior corners of 0.107 in length and 0.107 to 0.089 in width
the segment and measure1.874 to 1.339 in length and
6.366 to 5.473 in width. Testes are medium in size,
oval in shape, scattered all over segment, bounded RESULTS AND DISCUSSIONS
laterally by longitudinal excretory canals, in simple
field, in the central medulla, unevenly distributed, The genus Moniezia has been erected by Blanchard,
occupy the whole segment, 40 to 50(45) in number 1891 as type species Moniezia expansa (Rudolphi,
and measure 0.142 in length and 0.089 in width. The 1805) Yamaguti, 1942 from Ovis aries, Bos indicus,
cirrus pouch is situated near the anterior margin of the Capra capreolus etc. Later on many species are added
segment, medium in size, cylindrical in shape, curved, to this genus. After going through the literature, the
extends slightly beyond the longitudinal excretory worm under discussion comes under the subgenus,
canals, transversely elongated and measures 0.714 to Balanchariezia skrjabin and Schulz, 1937. The present
n0.679 in length and 0.142 to 0.035 in width. The worm, agree in the all the characterswith subgenus
cirrus is a thin tube, slightly coiled, slightly protruded, Blanchariezia Skrjabin and Schulz (1937) having two
contained within the cirrus pouch and measure 0.357 species as M. (B.) pallida monning, 1926. Later on two
in length and 0.035 in width. The vas deference is thin, more new species were added by Shinde et al., from
long, slightly wavy and measure 0.856 in length and the host Ovis bharal at Aurangabad, Maharashtra state,
0.035 in width. India. The present worm agree in all the characters of

Kale Sanjay S, 2017
genus moniezia and subgenus Blanachariezia, having The present cestode, differs from the M. (B.) bharlae
two species as M. (B.) pallida, Mooning, 1926, M. (B.) which is having, scolex not available, mature segments
benedieni (Moniez, 1879), Skrj. et Schulz, 1937. Later broder than long, testes 190-220 in number,
on two more new species were added to this genus by distributed in to of segment; ovary compact,
Shinde et, al., from the host Ovis bharal at Aurangabad bilobed; interproglottid glands in two rows, at the
as M. (B.) aurangabadensis and M. (B.) bharalae. posterior margin of segments and 38-44 in number;
seminal vesicle enolgated and fusiform; cirrus pouch,
The present worm comes closer to the following small, oval, obliquely placed; vitelline gland absent,
species: vagina posterior to cirrus pouch and host Ovis bharal
1. M. (B.) benedeni (Moniez, 1979) Skrj. et Schulz, (Sheep). By observing these distinct characters, the
1937. author has no other way, than to erect a new species,
2. M. (B.) aurangabadensis, Shinde et. Al., 1985 to accommodate these worm and hence the name
3. M. (B.) bharalae, Shinde et. Al., 1985 Moniezia (Blanchariezia) caprae n. sp. Is proposed,
after the gneric name of the host.
The worm difference from above species, in having the
scolex small in size, simple; suckers four in number, Type species -Moniezia (blanchariezia) caprae n.sp.
medium in size, oval in shape: neck long, mature Host - Capra hircus.
segment broader than long, with double set of Habitat -Intestine.
reproductive organs; testes medium in size, oval in Locality- Kasgi, Tq. Omerga, dist. Osmanabad,
shape, bounded by longitudinal excretory canals, in a Maharashtra state, India.
single field, unevenly distributed, 40-50 (45) in
number; cirrus pouch medium in size, cylindrical in Conflicts of interest: The authors stated that no conflicts of
shape; cirrus is thin, within the cirrus pouch; ovary interest.
bilobed, medium in size, lobes unequal, with irregular
margins; vagina posterior to the cirrus pouch, from REFERENCES
receptaculum seminis, opens in to ootype,
receptaculum seminis medium in size, oval in shape; Jadva BV (1985) Two new species of the cestode Moniezia
ootype small, oval; genital atrium large, oval, marginal; Blanchard 1881, Rivista parasitologia, XI 2(XLVV):1985,
PP-33-37.
genital pores are bilateral, small, oval; vitelline gland
Kale AT, Shinde GB (1999) Lytoceytus Chalisgaonensis n. sp.
medium, oval postovarian, compact and the Cestode Parasites from mammalian Riv. Parasit. 16 (60):
interproglottidal glands present in the intersegmental 1:39-42.
region, 12-13 in number, oval, arranged in paris, host Murhari et al. (1999) Parasites and Parasitosis of sheep.
Capra hircus (Goat). Indian journal Vel. Sci. Animal Husbandary 12:165 to
168.
The present tapeworm, difference from M. (B.) Patil SR, Shinde GB and Jadhav BV (1998) A new specues of
Genus Moniezia (B.) Paithensis M.S. Indian. Journal of
bendeni, which is having mature segments broader Parasitology Diseases, 22(2): 148-151.
than long, posterior segments fleshy; testes 500 in Pallid. 1926: Observation of worm burden of Goat from
number, arranged in the from of two triangles in two certain distericts of united provinces. Indian Jour. Vet.
fields; ovary compact, with acini, presentat at centre of Sci. 12: p-199-209
the segment on each side; interprporoglottid glands Skarjabin Schuz:1937: Noties heminthologies bull. Soc.
France 60:482- 501.
varying in size, narrow, short, and in a transverse row;
Thakare BG (2002) Ph. D. thesis Dept. of Zoology, Dr B.A.M.U.
cirrus pouch wide, short, oval, do not touch
Aurangabad.
longitudinal excretory canal; host horse. The worm
Yamaguti S (1959) Systemema Helminthum Vol. 1.2. Cestode
under discussion, difers from M. (B.) aurangabadensis, of vertebratesInterscience publ. new: ondan, 1-860pp.
wich is having scolex simple; segments broader than Yamaguti S (1934) Studies on helminthes fauna of japan
long; testes 1100-1200 in number, distributed fully, part-4Cestode in vertebrates.
from anterior to posterior of the segments; ovary Yang (2010) Seasional population dynamic of the cestode in
bilobed, each lobe with acini; inter proglottid glands wild life Helmithologia, 47, 2: 241-250, 2010. Japan
Journal of Zoology. 6:1-112.
12-15 in number, in a row; seminal vesicle oval and
large; cirrus pouch small, round; cylindrical vitelline
gland small, round; vagina posterior to cirrus pouch 2017| Published by IJLSCI
and host Ovis bharal (sheep).

RESEARCH ARTICLE
Sarus Crane in Indian Mythology and Culture
Kumar Adesh and Kanaujia Amita
Biodiversity & Wildlife Conservation Lab, Department of Zoology, University of Lucknow, Lucknow- 226007
Uttar Pradesh, India
Email: kanaujia.amita@gmail.com.
Received: 20.05.2017 The Sarus crane is known for its immense cultural contribution is the tallest
Accepted: 10.09.2017 flying bird and globally 'Vulnerable' species as per IUCN Red List of
Published : 30.09.2017 Threatened Species. It is the state bird of Uttar Pradesh of India. The
cultures of various communities not only in India but from all over the
Editor: world have mentioned about Sarus such as Nepal, Vietnamese, Chinese and
Dr. Arvind Chavhan Cambodia. Many myths are associated with Sarus. This bird is also known
as the eternal symbol of unconditional love and devotion and good fortune.
Sarus has a special position in social folklores and this is reflected in its
Kumar Adesh and Kanaujia Amita
mention in some Indian folk stories and songs. From the ancient to modern
(2017) Sarus Crane in Indian
Mythology and Culture; International time many painters, artists and workers are concerning the Sarus crane in
J. of Life Sciences, 5 (3): 465-470. their paintings, architectures, embroideries, handicrafts. India Post also
launched a postal stamp on Sarus crane. Presence of Sarus crane is good
sign of rainfall and healthy wetlands ecosystem. Various Government
Acknowledgements departments, Universities, Scientists and Conservationists are coming
Thank are due to, Head Department together to work for Sarus conservation which includes both ex-situ and in-
of Zoology, University of Lucknow, situ measures. Since we are depleting our wetlands which direct affect the
Lucknow for providing facilities for
Sarus day by day, World Wetlands Day (2nd February) is being celebrated all
study. Authors thanks to all the
over the world to make the future generation sensitive about the dilemma
volunteers, local peoples and
traditionally knowledgeable persons of Sarus.
for their continuous help and
support. Key words: Sarus crane, Mythology, Culture, Symbol.

INTRODUCTION
and distribution in any medium, Our ancient civilizations depict the worship of some of animals and birds
provided the original work is including nature Gods. Even the forests, environment and the elements of
properly cited, the use is non- nature were revered and held in Indian Mythology with high esteem. Sarus
commercial and no modifications or crane known as the eternal symbol of unconditional love and devotion and
adaptations are made. good fortune in Indian culture. Due to developmental activities and
population pressure the population of this bird is depletion day by day. The
Sarus crane which is known for its immense cultural contribution is the
tallest flying bird and globally 'Vulnerable' species as per IUCN Red List of
Threatened Species. It is the state bird of Uttar Pradesh of India. The Sarus
Crane (Grus antigone) is a large non-migratory crane. The name Sarus came

Kumar and Kanaujia, 2017
from Sanskrit term sarasa which means bird of the associated with Sarus within India and the World. Data
lake. All the way through the ages, Sarus have been was also collected during visits to various places like
honored as symbols of love and devotion. All through Lumbini Garden in Nepal, Bodh Gaya in Uttar Pradesh,
olden times and across many cultures, people have Bodh Stoop in Banaras.
regarded Sarus with charm and respect.
Sarus in Indian Mythology and Culture
Sarus have important place in mythology and culture The Sarus crane species has historically been
along with their economic significance and ecological distributed broadly in the northern and north-western
role as iconic bird of Indian wetlands. lowlands of India along the plains of the Ganges River.
From the ancient time it is widely believed that the In India, the Sarus crane are considered sacred birds;
Sarus pairs for life and that death of one partner leads they never get harmed by human. The Sarus crane has
to the other infatuated to death. Assumption about always had a privileged social status in India. Many
Sarus crane began in primitive folklores, ages ago but myths are associated with this bird. This bird is also
passed over to the future generations through known as the eternal symbol of unconditional love and
narratives and tales. Sarus crane notorious as the devotion and good fortune.
eternal symbol of unconditional love, devotion,
togetherness and good fortune in Indian culture and There is a tradition of having a newly-wed couple view
different myths. The cultures of various communities a Sarus pair in some parts of India. It is believed that if
not only in India but from all over the world have one bird of the pair were to die, then the other
mentioned about Sarus such as Nepal, Vietnamese, shadows the path towards starvation by refusing any
Chinese and Cambodia. The purpose of this paper is to food or water (Sunder & Chaudhary, 2003). Sarus has
imitate the magnitude of the most efficient iconic bird a special position in social folklores and this is
of wetlands throughout the ancient times in India. reflected in its mention in some Indian folk stories and
There have been detailed studies regarding the role of songs (Mishra, 2009).
Sarus crane in ecosystem, this paper reflects their role
in various cultures. Sarus pairs are a symbol of marital virtue and in parts
of Gujarat and believed that death of one partner leads
to the other infatuation to death. The mating season of
METHODOLOGY sarus coincides with two of the most important
cultural occasions in the Indian subcontinent: The
To be acquainted with the significance of the Sarus in Dashain and the Tihar. These back to back celebration
numerous cultures around the World, all existing events are festive and cheerful and very significant to
scientific and grey literature was sited and reviewed the people, analogous to our traditional Lunar New
on the mythological, cultural, occurrence and rituals Year.
Fig.1 Valmiki cursed a hunter for killing a Sarus crane

The sarus is recognized in India and legend has it that between them and that the incubation period was 34
the poet Valmiki cursed a hunter for killing a Sarus days in floods plains of Ganges (Johnsgard, 1983).
crane and was then motivated to write the epic
Ramayana (Rana and Prakash, 2004, Sunder and In 19th-century British zoologist Thomas C. Jerdon,
Chaudhary, 2005) (Fig.1). consulted that young birds were good to eat, whereas
older ones were "worthless for the table" (Tanee et al.,
The tribes of Gondi people consider sarus crane as 2009). In some parts of India, the eggs of the sarus
sacred five god worshippers (Birdlife International, crane are still used in traditional remedies (Leslie,
2001). In ancient Hindu scriptures the meat of sarus 1998 and Sundar & Chaudhary, 2003).
was considered taboo (banned or unmentionable,
Board et al., 2001). A plate of Sarus crane by Johann Michael
Seligmann published between 1749 and 1776 based
Around 1607, observation on sarus biology made by on a work by George Edwards (Fig.2). Maharana Amar
the Mughal emperor Jahangir and concluded that sarus Singh Hunting Sarus crane have shown in fig-3.
constantly laid two eggs with an interval of 48 hours
Fig.2 A plate of Sarus crane used by J M Seligmann Fig.3 Maharana Amar Singh Hunting Sarus crane
Fig.4 Sarus crane in Handicrafts Fig.5 Sarus crane in Paintings

Fig.6 Sarus crane in Embroidery
Fig.7 Sarus crane in Postal Stamps
Sarus crane also portrays in Indian Handicrafts (Fig.4), A few historical interpreters of Buddhist transcript
Paintings (Fig.5), Embroidery (Fig.6) and Postal from Chinese to Vietnamese have revealed a story of
Stamps (Fig.7). the young Prince Siddhartha (before he became the
Buddha) and the swan (Fig.8 & 9). When the prince
Sarus crane in other Culture of World was very young, one day he found an injured swan
(some of assumed that was sarus) on the palace
The term Sarus came from Sanskrit term sarasa
ground that was shot by his cousin Devadatta.
which means bird of the lake. The very earliest
Siddhartha detached the poisoned arrow, and nursed
pioneers of the Indian subcontinent region speak the
the wounded bird with extreme sympathy. When
Austro-Asiatic language, sarasa is derived from there
Devadatta found out that it was the bird he shot down,
which is also the root language of most Asian nations
he wanted to own the prize bird and demanded it
including the old Vietnam. Vietnamese people called
back. Heated argument succeeded. Siddhartha won the
these cranes seu but in Vietnamese folks the
argument by proving that the life of an animal belongs
Chinese-derived name hac that was only poetically
to the person who rescued it, and not to the one who
applied in written literature.
dispossessed it.

Fig.8 Sarus in Vietnamese Culture Fig.9 Prince Siddhartha with Sarus
Fig.10 painting by Melchior dHondecoeters (1680) Fig.11 wheel of sarus crane conservation depicts
showing a sarus crane in the background the life cycle, threats and conservational measures
for sarus conservation and protection

But according to the intellectual scholar Hemraj REFERENCES

Shakya - who was the most proficient in ancient
Birdlife International (2001) Threatened birds of Asia: The
languages like Brahmi, Sanskrit and Pali, and who
BirdLife International Red Data Book. (PDF). BirdLife
signified the most admired of all researchers of ancient International, Cambridge, UK. ISBN 0-946888-42-6.
Indian and Nepalese history there are no Blaauw FE (1897) A monograph of the cranes. Leiden and
acknowledged species of swans or flying geese in said London. 22 plates +64pp.
Board CK, Mukherjee A and Parasharya BM (2001) "Damage
territory at the time of Buddha. The bird in potential of Indian sarus crane in paddy crop
Siddharthas story must be the native Sarus crane, agroecosystem in Kheda district Gujarat, India".
whose marshland habitat distribution throughout Agriculture, Ecosystems and Environment, 86 (2): 211
215.
India is more relevant of Sakyamuni Buddhas country
Johnsgard PA (1983) Cranes of the world. Indiana University
(Nguyen Phu, 2011). Press, Bloomington. ISBN 0-253-11255-9.
Leslie J (1998) "A bird bereaved: The identity and
significance of Valmiki's kraunca". Journal of Indian
The floating feather: a painting by Melchior Philosophy, 26 (5): 455487.
dHondecoeter (1680) of the birds in the menagerie of Mishra AK (2009) - Sarus crane: on its way to extinction.
William III of England at the Het Loo Palace showing a Resonance. pp 1206-1209.
Phu N (2011) Lumbini- Land of Sarus crane.
sarus crane in the background worked named Het
Rana G and Prakash V (2004) "Unusually high mortality of
drijvendvveertje (Blaauw, 1897) (fig.10). In Pakistan, cranes in areas adjoining Keoladeo National Park,
sarus cranes are apparently cited in poetry (D. Bharatpur, Rajasthan". J. Bombay Nat. Hist. Soc. 101 (2):
Ferguson, in litt., 2003). 317.
Sundar KSG and Choudhury BC (2003) "The Indian Sarus
Crane Grus a. antigone: a literature review". J. Ecol. Soc.
A Nepalese Traditional Thanka painting based on 16: 1641.
Buddhist wheel of Life Philosophy which show the Sundar KSG and Choudhury BC (2005) "Mortality of sarus
cranes (Grus antigone) due to electricity wires in Uttar
wheel of sarus crane conservation and depicts the life
Pradesh, India". Environmental Conservation. 32 (3):
cycle, threats and conservational measures for sarus 260269.
conservation and protection (Fig. 11). Tanee T, Chaveerach A, Anuniwat A, Tanomtong A, Pinthong
K, Sudmoon R and Mokkamul P (2009) "Molecular
Analysis for Genetic Diversity and Distance of
Introduced Grus antigone sharpii L. to Thailand".
CONCLUSION Pakistan Journal of Biological Sciences. 12 (2): 163167.
Sarus crane notorious as the eternal symbol of

unconditional love, devotion, togetherness and good 2017| Published by IJLSCI
fortune in Indian culture and different myths. From
the ancient to modern time many painters, artists and
workers are concerning the Sarus crane in their
paintings, architectures, embroideries, handicrafts.
Presence of Sarus crane is good sign of rainfall and
healthy wetlands ecosystem. Various Government
departments, Universities, Scientists and
Conservationists are coming together to work for
Sarus conservation which includes both ex-situ and in-
situ measures. Since we are depleting our wetlands
which direct affect the Sarus day by day, World
Wetlands Day (2nd February) is being celebrated all
over the world to make the future generation sensitive
about the dilemma of Sarus.

interest.

RESEARCH ARTICLE
Burud Society and Bamboo Plants Utility : Dendrocalamus strictus
Deshmukh Pratap V
Department of Botany, Nagnath Arts, Commerce and Science College, Aundha Nagnath Dist- Hingoli, MS, India.

Received: 13.08.2017 Peoples from Burud society are economically poor and there is no any
Accepted: 23.09.2017 another source for livelihood. From their ancestors they weaves some
Published : 30.09.2017 things useful for daily needs from plant material like Lantana, Ipomoea,
Shindi etc. After some working experience they leads for manufacture of
Editor:
articles from Bamboo. Some plants are very soft and some are very strong.
Dr. Arvind Chavhan
Out of these plants one was bamboo which is very high and stem is
Cite this article as: strength. In our India Bamboo plants are distributed I Andhra Pradesh,
Deshmukh Pratap V (2017) Burud Kerala, Karnataka, Himachal Pradesh, Maharashtra etc. This plant is very
Society and Bamboo Plants Utility : useful and maintains diversity of flora of region. No of peoples and their life
Dendrocalamus strictus; style is depends on such plants because they got currency and job for daily
International J. of Life Sciences, 5 maintenance. Dendrocalamus strictus Bamboo have no of uses to human
(3): 471-473. being and also to environment.
Acknowledgement: Key words: Ancestors, livelihood, Forest, Strength, Flora, Diversity, etc.
I am thankful to my management
members of my institute. I am
grateful to my Honorable Principal. I INTRODUCTION
also express my sensior thanks to
Burud society peoples and my In Burud society families are in large sized. In this family one member is
couliges.I am heartily thankful to my
head and others cooperate to him for weaving work. At ancient time they
students.
collect Bamboo plants from forest and other regions like river bank, dam,
Copyright: 2017| Author (s), This and streams. We are very familiar with Bamboo which occurs in tropical
is an open access article under the and subtropical region of India. I n India Bamboo species are about 22 and
terms of the Creative Commons subspecies are100-120, out of them 50% Bamboo is present in East India.
Attribution-Non-Commercial - No Bamboo grow in plane, hilly and water perforated land. Bamboos family is
Derivs License, which permits use Poaceae or Gramineae. The Bamboo plant is about 25-50 feet in height, and
and distribution in any medium, stem is divided in to nodes and internodes. Growing buds are present on
provided the original work is nodes which grow later on in branches. Root system is adventitious and run
parallel to the earth. Roots conserve the soil by tight holding, also increase
the organic material and porous nature. Leaves have diversity, flowers are
small and in group. After seed formation Bamboo plant will died (after 40-
60 years). Dendrocalamus strictus Bamboo is such a plant which is known to
everyone who wanders in the forest. But now a day this plant is observed in
the farm field also. No of farmers doing agriculture of Dendrocalamus
strictus bamboos plants as a crop production. Dendrocalamus strictus
Bamboo plant is useful to human being in social as well as financial role. It
may be called as gold crop due to its reach value. It plays a financial role in
India, China, Japan hence it is important crop. Bamboo is useful from birth
to death of human being.

Deshmukh Pratap V, 2017
MATERIALS AND METHODS anybody can do the marketing. Hence in India about
one lakh peoples get job.
In Burud society families are in large sized. In this
family one member is head and others cooperate to After Chin, India is second bamboo production country
him for weaving work. At ancient time they collect and our Maharashtra is at 5th level for production.
Bamboo plants from forest and other regions like river Westernghat, north Maharashtra, Gadchiroli,
bank, dam, and streams. We observe different forest Chandrapur, Gondia are the main regions for
then we look towards stem crowded tree which is very Dendrocalamus strictus bamboo production. I n this
high and somewhat green in colour. When we survey region scientific technically cultivation is done.
the fields in different study tours, then we collected
the specimens of no of plants. In this group Environmentally bamboo is very important, roots of
Dendrocalamus strictus bamboo is also collected, dried. this plant are fibrous and anchorage to the soil tightly,
Mounted on herbarium sheet and preserved stored in hence soil should not flow with current of water flow.
herbarium cabinet of our Botany Department of Hence it help in conservation of soil. Help in soil
Nagnath Arts, Commerce and Science College, Aundha erosion along the river of banks. It helps in carbon
Nagnath Dist- Hingoli. Which is useful for study to absorption and increase fertility of soil. It is also
research students. cultivated in barren land, low land. Rhizome grows
horizontally and new plants are grown. It grows on
bunds of field and acts as hedge for crop in fields. In
RESULTS AND DISCUSSIONS our India, there are about 90 genera and 150 pieces.
For identification of species characters are useful like
In whole world the Bamboo production is more in length and girth of internodes, sheathing leaf base and
China and secondly in our India. In China 20% area spines growth. In Maharashtra Dendrocalamus strictus
occupied by Bamboo and in India it is about 13%. On Bamboosa is a familier species.Boorude society is
the basis of environmental balance, development of totally depends on these plants for daily life. They
Bamboo forest as a production source is our need. In manufacture different types of things like basket,
Maharashtra Amravati district Wadali Bamboo showpiece, and sold in market.
nursery is famous. In this region different types of
Bamboo species are collected. Now a days in Konkan, Utility of Bamboo:
Kolhapur and Pune district Bamboo plantation is in In forest on the road side Bamboo plantation is
progress. In Maharashtra Bamboo species like Kanak, necessary to avoid accidental cases.High quality paper
Manga, Mesh, Baramesh, Chiva found in large scale. is also made from Bamboo plant. Bamboo is also used
in Agarbatti preparation. Dendrocalamus strictus
Dendrocalamus strictus Bamboo is a Bamboo is highly useful in manufacturing Suplee,
monocotyledonous plant which comes in family Topli, Sidi. Agricultural equipments-rod for diggers
Gramineae or Poaceae. This plant is occurring on (Tikav, Fawada), Balad, Kangi (Grain storage),
bunds of field, on road side and on bank of river, in supporter for orange, grapes, tomato, banana farm. In
forest etc. It is green deciduous plant. It is tall tree, our house it is useful as suplee, topli (Basket), put
roots are adventitious fibrous, steelt root may be bread or roti in this topli in kitchen. Carriers like
present. Stem is long and divided in to nodes and bulackart, ship. hodi, nav. In house building like walls,
internodes on the basis of length of internodes species roof, partitions, doors, windows etc. Furniture like
differentiation can be done. At nodal region leaves table, chairs, sofa, teapot Commercially Dendrocalamus
arises with sheathing leaf base leaf lamina is elongated strictus bamboo is useful in packing fruits, glass
with parallel venation and trichomes on the surface. material, flowers etc. It is used in manufacture for
Bamboo is useful to human being from birth to death. sticks of agarbatti, frame. Bhala, dhanusheban, Lathi. It
It is cash crop for agriculturist. It is precious for giving conserve the soil and increase the fertility of soil.
job opportunity. In different sectors Dendrocalamus Medicinally it is useful on Naru disease.
strictus bamboo is useful such as Building project, Dendrocalamus strictus Bamboo are used in making
paper project, toys projects, different showpieces etc. tree guards to protect the plants from other animals.
In ancient period Boorude society work on this On the occasion of Pola festival of farmers food is
Dendrocalamus strictus bamboo plant but now a day given to oxes in topli,supli of Dendrocalamus strictus

Burud Society and Bamboo Plants Utility : Dendrocalamus strictus
All these things like Topli(Basket), Suplie, Sidi, Mat, Sticks, Treeguard etc are made by Burud society peoples.
bamboo and invite them. It has a loading capacity hence for building purposes hence it gives inspiration for
it is used for construction of houses, ship, shed house, plantation.
greenhouse etc. It is also used for making ply board and
paper. Bamboo plant is not useful to poor, middle class Conflicts of interest: The authors stated that no conflicts of
people but also to rich peoples. The species like Bima interest.
Bamboo is useful in electricity production, such type of
electricity is produced from Bima Bamboo in south
Africa. It is also possible in our India for that project REFERENCES
should be developed. Bamboo parts like leaves, flowers,
Cooke T (1901-1908) The Flora of the Presidency of
buds medicinally useful on cough, acidity and
Bombay London. 2 vols: Repr. edition, 1958, B. S. I.
rheumatism. Bamboo is applicable in abdominal pain Calcutta.
treatment and in some gynic diseases. Pickles are made Hooker JD and Thomson T (1855) Flora India. London Vol.
from young branches of Bamboo after boiling. 1.
Singhs NP, Lakshminarsimhan P and Prasanna SK (2000)
Flora of Maharashtra Sts. Dicotyledons. Vol. 2. BSI,
Western Circle, Pune.
CONCLUSION:
Ketkar SM and Pandit Shrivanas () Bamboo: Srusti and
Drusti,Sakal Prakashan.
Burud society peoples are most important in our
Lakshminarsimhan P and Prasanna S (2001) Flora of
nation. Due to them importance of Bamboo plant is Maharashtra Sts. Dicotyledons vol.1 by N. P. Singh and
spread all over the world. Hence we may say that karthikeyan. BSI, Western Circle, Pune.
Bamboo is a Green Gold. Dendrocalamus strictus Naik VN (1969) The Flora of Osmanabad. Aurangabad,
Bamboo is an important tree for human being and Venus Publishers.
environment in different way hence it should be Naik VN (1998) Flora of Marathwada Vol. I & II Amrut
Prakashan, Aurangabad.
cultivated, grown and conserved carefully. It maintains
Vishnu Swarup (1967) Gardens Flowers. National Book
the diversity of nature and ecological balance. Today we Trust, India.
are looking to words that plantation of bamboo is done Yadav SR and Sardesai MM (2002) Flora of Kolhapur
in all over the country. When we think about District. Shivaji Univresity, Kolhapur.
Maharashtra, in Konkan, Viderbha, on hill tops
Dendrocalamus strictus bamboo have tremendous 2017| Published by IJLSCI
scope. Today Dendrocalamus strictus bamboo is used

OVERVIEW
Assessing the impact of pesticides: An overview
Jawale CA1, Rajput KH1 and Ugale BJ2
1Department of Zoology, Shri Madhavrao Patil Mahavidyalaya, Murum, Tq: Omerga, Dist. Osmanabad (MS)India.
413605.
2Department of Zoology, Shrikrushna Mahavidyalya, Gunjoti. Tq: Omerga, Dist. Osmanabad (M.S.) India. 413613

Received: 21.07.2017 There are three types of pesticides. They are organochlorine,
Accepted: 21.09.2017 organophosphate and carbomate compounds. Organophosphorus
Published : 30.09.2017 compounds are remarkable for their selective toxicity, low persistence and
greater biodegradability. Organophosphates are less lipophilic than
Editor:
chlorinated hydrocarbons. Because of the rapid biodegradability, the
Dr. Arvind Chavhan
organophosphate insecticides have replaced more persistent
Cite this article as: organochlorine compounds and are used in greater quantity. India accounts
Jawale CA, Rajput KH and Ugale BJ for one third of pesticides poisoning case in the world. A majority of them
(2017) Assessing the impact of get exposed for long periods cases of blindness, cancer, deformities,
pesticides: An overview; diseases of liver and nervous system etc., from pesticide poisoning have
International J. of Life Sciences, 5 (3): been identified. These pesticides however have a large scale of
474-479. contamination in the ecosystem as a result of which they cause various
effects on the biotic components of the environments besides the pests or
targets species for which the chemical is applied. The pesticide across
Acknowledgement:
media indicates portals of entry into the existing food web. The basic
The authors are thankful to
Principal, S.M.P. Mahavidyalaya, principle of adsorption, distribution, metabolism and excretion all
Murum. Dist. Osmanabad for toxicology is equally important when assessing toxic effects of pesticides on
providing necessary facilities, animal life. Various animals have also been used as indicators to measure
valuable suggestions and the extent of pesticide pollution of the environment. The present
encouraging carry out the investigations are attention is given mostly to studies pesticide effects on
present research work.
animal life and the environments.
Copyright: 2017| Author (s), This Keywords: Pesticides, Toxicity, Health hazards and animals.
Derivs License, which permits use INTRODUCTION
provided the original work is Pesticides are widely used in agricultural production to prevent or control
properly cited, the use is non- pests, diseases, weeds and other plant pathogens in an effort or reduce yield
commercial and no modifications or losses and maintain high product quality (Abbasi and Krishnan, 1993).
adaptations are made. Pesticides are commonly used to protect animals from pests. However, it is
important to remember that any pesticides should be considered an active
poison. For sustainable agriculture and protection of the environment and
human health, the importance of using the pesticides has assumed global
importance subsequent to in Earth Summit in 1992. Although pesticide are
developed through very strict regulation processes to function with

Assessing the impact of pesticides
reasonable certainty and animal impact on human protect plants from weeds, fungi or insects. Pesticide is
health and the environment, serious concerns have a chemical or biological agent that deters,
been raised about health risks resulting from incapacitates, kills or otherwise discourages pests.
occupational exposure and from residues in food and Target pests can includes insects, plant pathogen,
drinking water. Pesticide uses has raised serious weeds, mollusks, birds, mammals, fish and microbes
concern not only of potential effect on human health, destroy property or disease vector and the
but also about impact on wildlife and sensitive environments are fully justified (Gilden et al. 2010;
ecosystem (Kamrin, 1997). Tariq et al., 2007).
The World Health Organization estimates that there The purpose of this present paper is to review of
are three million severe acute poisoning world wise literature assessing the safety, impact of pesticides on
each year and out of this, approximately 2,20,000 human health and other animal life the environment
death are attributable pesticides, out of which, 1% of effects.
these death occurs in industrialized countries (Sataka
et al., 1997). The pattern of pesticide usages in India is HISTORY OF PESTICIDES:
different from that of the world in general. In India
76% of the pesticides are used as insecticide, as The first known pesticide was elemental sulfur dusting
against 44% globally. The use of herbicides and used in ancient Sumer about 4500 years ago in ancient
fungicides are correspondingly less. The main use of Mesopotamia. The Rig Veda, which is about 4000 years
pesticides in India is for cotton crops 45%, followed by old, mentions the use of poisonous plants for pest
paddy field and wheat farming (Relyea and Dieck, control (Rao et al., 2007). In the 17th century, nicotine
2008). The use of pesticides in tea and vegetable crops sulfate was extracted from tobacco leaves for use as an
in India is not negligible. Moreover, repeated low dose insecticide. In 19th century saw the introduction of
application has caused large impact in the agro- natural pesticides, which is derived from the roots of
ecosystems than single exposure (Yassi et al., 2001; tropical vegetable (Miller, 2002). And first important
Manchini et al., 2005). synthetic organic pesticide DDT was discovered in
1939 by Swiss chemist Paul Muller. DDT was a very
At present, India is the largest producer of pesticides effective insecticide but they were replaced in the US
in Asia and ranks XIIth in the world for the use of by organophosphates and carbonates by 1975 (Ritter,
pesticides with an annual production of 90, 000 tons 2009).
(www.teri.res.in/peticide.htm). A vast majority of the
population (56.7%) in India are engaged in agriculture The U.S. Environmental Protection Agency was
and are therefore, exposed to the pesticides used established in 1970 and amendment to the pesticide
(Gupta, 2004; Government of India, 2001). Pesticides law in 1972 (US, EPA; 2011). Pesticides use has
that are being used in agriculture fields disseminate increased and 2.3 million tons of industrial pesticides
into the environment and come in human contact are now used each years (Miller, 2002). The 75% of all
directly or indirectly. Human are exposed to pesticides pesticides in the world are used in developed
that are found in the environmental by different routes countries, but use in developing countries is
of exposures like inhalation, ingestion and dermal increasing. In recent years, chemical pesticides have
contacts, keeping these in the backdrop among the become the most important consciously applied form
farmers. Pesticides are chemicals that are meant to of pest management. This is a generalization of course
kill (Van der, 1996). The pesticides includes all of the crops in same area. Pesticides are generally more
following herbicide, insecticides, insect growth water soluble and are often also more acute toxic.
regular, nematicide, termiticide, molluscicide, Pesticides effects on animal life such a fish, birds,
piscicide, avicide, rodenticide, predacide, bactericide, reptiles, insects, mosquitoes etc., and which was a
insect repellent, animal repellent, antimicrobial, serious threat to biodiversity (Golden, 2007).
fungicide, disinfectant antimicrobial and sanitizer. The
most common of these are herbicides which account DEFINITION OF PESTICIDES:
for approximately 80% of all pesticide use (Carolyn
Randall ed, 2013). Most pesticides are intended to Pesticides are unique among toxic substances. Most
serve as crop protection products, which in general environmental toxins are an unwanted by product of

Jawale et al., 2017
another process. Pesticides are chemicals designed to that regulates acetylcholine, a neurotransmitter. Most
be harmful to a target pest and purposely introduced organophosphates are insecticides. They were
into the environment to do their job managing insects, developed during the early 19th century, but their
bacteria, weeds, rodents or other pests (US, Envoi, effect on insect, which are similar to their effects on
2007). human, were discovered in 1932. Some are very
poisonous and they usually are not persistent in the
Pesticide means insecticides. Pesticide refers to not environment.
only insecticides but many other kinds of chemicals.
Under state and federal law, a pesticide is any Organochlorine insecticides:
substance intended to control, destroy, repel or attract They were commonly used in the past, but many have
a pest (Kamrin, 1997; Goldman, 2007). been removed from the market due to their health and
the environmental effects and their persistence (DDT)
TYPES OF PESTICIDES chlordane and taxaphene.
Pesticides are often referred to according to the type Carbamate pesticides:

of pest they control. Pesticides can also be considered Carbamate pesticides affect the nervous system by
as either biodegradable pesticide. They will be broken disrupting an enzyme that regulates acetylcholine, a
down by microbes and other living beings led to its neurotransmitter. The enzyme effects are usually
accumulation in the food chain and its killing of living reversible. There are several subgroups within the
animals of prey at the top of the food chain. Another carbamates.
way to think about pesticides is to consider those that
are chemical pesticides or are derived from a common Pyrethroid pesticides:
source or production method. Various pesticides were They are recently developed as a synthetic version of
as given in the Table -1. the naturally occurring pesticide pyrethrin, which is
found in chrysanthemums. They have been modified to
Organophosphate pesticides: increase their stability in the environment. Some
Organophosphates affect the nervous system by synthetic pyrethroids are toxic to the nervous system.
disrupting, acetyl cholinesterase activity, the enzyme
Table-1: Various types of the pesticides for Agriculture field use in safe concentration.
Sr. Types of the Pesticides Name of the Pesticides Safe concentration of the
No. pesticides in agriculture field
01 Organophosphate Monocrotophos 0.04 to 0.06
Dimethoate 0.001 to 0.003
Choloropyriphos 0.001
Dichlorovos (DDVP) 0.001
Phosphamidon 0.003
Phosalone 0.004
Malathion 0.001
Endrin 0.0001to 0.0002
02 Organochloride Endosulfan-I 0.001
Endosulfan-II 0.002
DDT 0.001 (2 kg/ha)
03 Carbamate Carbofuran 0.001- 0.009
Carbosulfan 0.001
04 Synthetic pyrithroids Permethrin 0.001- 0.003
Cypermethrin 0.001-0.002

Fig -1: Indicates of the bottle contains pesticide potential.
Neonicotinoid pesticides: pesticides are classified into four categories as

Neonicotinoids are a class of neurons active indicated by their label, such as Red as indicates
insecticides chemically similar to nicotine. It is the extremely poisonous, Yellow indicates poisonous,
mostly widely used insecticide in the world Green indicates moderately poisonous and Blue
(Yamamoto, 1999). In the late 1990 neonicotinoids indicates low poisonously. It is naturally occurring
came under increasing scrutiny over their substances that control pest and microbial diseases.
environmental impact and were linked in a range of Have significance about potential pesticide exposure.
studies to adverse ecological effects, including honey Many pesticides under red label are banned now.
bee colony collapse disorder and loss of birds due to a Label of the pesticides are Red, Yellow, Green or Blue
reduction in insect population. In 2013, the European based on their order of decrease in poisonous effect to
Union and a few non EU countries restricted the use of animals or plants (Fig-1).
certain neonicotinoids pesticides (Cressey, 2013).
USES OF PESTICIDES / INSECTICIDES:
Herbicides:
It has been commercialized for weed control. These are useful for both agriculture and human
Chlorsulfuron are broad spectrum herbicides that kill purpose. Plant pesticides increase crop yield in
plants by inhibiting the enzyme acetoacetate synthase. agriculture by protecting plants. Pesticide use also
In the 1960, more than 1 kg/hac (0.89 Ib/Acre) crop controls spread of diseases by fungi, bacteria, insects
protection chemical was typically applied, while other from one region of farms to other region or even
herbicides sulfonylureas, sulfosulfuron, etc. allow as countries. It was seen that some pests which were not
little as 1% as much material to achieve the same present came here due to import of wheat or other
effect (Lamberth et al., 2013). countries. So the insect or bacteria if controlled,
diseases spread in between countries or places can be
Herbal pesticides (Bio pesticides): prevented. Pesticides used by the farmers are
It is also known as Bio pesticides. Herbal pesticides are extremely hazardous, many are highly and few are
certain types of pesticides derived from such natural slightly hazardous. The farmers also used some other
materials as animals, plants, bacteria, fungus or products like ash, cow dung, oilcake, kerosene and
viruses and minerals. Their use is controversial, lime dust, Neem leaves, Neem cakes, mixture of
especially in many European countries (Coombs, Amy, tobacco leaves and washing indicum etc., in their farm
2013). land. But the use of these products were in very less
quantities as compared to their inorganic parts
The pesticide, Insecticides differently is not seen in (Hoppin et al., 2002; Horrigan, et al., 2002; Ecobichon,
term of their toxicity they are equally toxic. The 2001).

Jawale et al., 2017
POLLUTION OF PESTICIDES: pesticide movement but also may reduce

bioavailability to microbial degraders (Sims and
Pesticides and insecticides are some of the most toxic Cupples, 1999).
chemicals used in present world on regular basis.
These are chemical agents used to kill pests or plants. Pesticides toxicity has hazardous effects on plants, soil,
The environmental effect of pesticides is a concern human, birds and other animals. The effect is
now. These pesticides are spread into air, water bodies poisonous and many physiological changes (system)
like rivers, canals, soil due to their regular use in the are affected by them. Due to pollution of soil, air and
form of spraying, powders, solutions in agriculture. In water these chemicals and pesticides get into the body.
homes and garden when used as mosquito repellents, Pesticides toxicity can cause a variety of adverse
bed bug spray etc., in homes or regular basis enter into health effects, ranging from simple irritation of the
physiology of humans and animals on breathing. Those skin and eyes to more severe effects such as affecting
food grains grown in pesticide polluted soil enter the the nervous system, mimicking hormones causing
food chain of all the living animals. The pesticides reproductive problems and also causing cancer (US,
entering water resources travel to other places EPA, 2006). A 2007, systematic review found that most
spreading the pollution of these pesticides (Honsby et studies on lymphoma and leukemia showed positive
al., 1993; Paul, 2002). associations with pesticide exposure (Bassil et al.,
2007). There is substantial evidence of associations
IMPACTS OF PESTICIDE AND INSECTICIDE: between organophosphate insecticide exposures and
neurobehavioral alterations. Limited evidence also
Pesticides use raises a number of environmental exists for other negative outcomes from pesticides
concerns. Over 98% of sprayed insecticides and 95% exposure including neurological, birth defects and fetal
of herbicides reach a destination other than their death (Mink et al., 2011).
target species, including non-target species, air, water
and soil. Pesticide drift occurs when pesticides The World Health Organization (WHO) UN
suspended in the air as particles are carried by wind to environment program estimate that each year, three
other areas, potentially contaminating them. Pesticides million workers in agriculture in the developing world
are one of the causes of water pollution and some experience severe poisoning from pesticides, about
pesticides are persistent organic pollutants and 18,000 of whom die, and 99% of pesticide related
contribute to soil contamination. Biological deaths occur in developing countries that account for
magnification is the process were by these pesticides only 25% of pesticide usages (Miller, 2004). According
are more concentrated at each level of the food chain to one study, as many as 25 million workers in
(Nwilene et al., 2008). Among marine animals, developing countries may suffer mild pesticide
pesticides concentration are higher in carnivorous poisoning yearly. There are several pesticides carvers
fishes, even more so in the fish eating birds and a side from agriculture that may also put individuals at
mammals at the top of the ecological pyramid (Castro risk of health effects (Jeyaratnam, 1990).
peter and Michael, 2010). Global distillation is the
process were by pesticides are transported from It is concluded that the previous literature, farmers
warmer to colder regions of the Earth, in particular the experienced a variety of significance and symptoms
poles and mountain tops. Pesticides that evaporate related to pesticides. Among the world populations,
into the atmosphere at relatively high temperature can the prevalence of signs and symptoms related to
be carried considerable distance by the wind to an pesticide exposure was higher among the farmers
area of lower temperature where they condense and involved in spraying. The higher percentage of some
are carried back to the ground in rain (Atreya, 2007). signs and symptoms among the sprayers could be due
It is desirable that pesticides be degradable or at least to their direct exposure to pesticides or due to
quickly deactivated in the environment. Such loss of previous exposure to pesticides. It can thus be
activity or toxicity of pesticides is due to both innate suggested that due emphasis is required to be given on
chemical properties of the compounds and the adoption of protective measures among the
environmental processes or condition. The presence of farmers and all agricultural field of the global in
chemical structure often slows down degradation in an general. Pesticides are developed to function with
aerobic environment. Absorption to soil may retard reasonable certainty and minimal risk to human health

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sustainable agriculture can address the environmental

Jawale et al., 2017
Author Index
Abah J 297 Nigussie Dechassa 345

Beyenesh Zemichael 345 Nikam S 425
Chandra Naresh 379 Pande Shishir 425
Chavan PN 459 Parashare VM425
Cherukuri V Raghavaiah 311 Parbhankar RL 447
Das Debangshu N 362 Parvathi D399
Das Tapati 362 Patil Avinash 379
Deshmukh Pratap V 471 Patil RB 420
Deshmukh RS 394 Patil RN 409
Gebretsadkan Zereabruk 323 Pawar PV 420
Gogoi Budhin 362 Pokale SS 438
Hay CJ 297 Rajender Reddy A 399
Jadhav BV409 Rajput KH 474
Jawale CA 474 Ramya K 387
Kachari Akash 362 Safi Vivekanand 362
Kadam Pratima S 405 Saikia Rajashree 362
Kale SS 455 Sajjan MB 409
Kale Sanjay S 462 Sammer Marzouk 291, 306, 332
Kanaujia Amita 465 Selamyihun Kidanu 311
Krishna Reddy V 399 Shinde VA420
Kumar Adesh 465 Simasiku EK297
Mafwila SK 297 Tamlurkar HL 451
Mahajan VS438 Thirunalasundari Thiyagarajan 387
Mali RP 459 Totawar DV 451
Matengu KK 297 Ugale BJ 474
Mogle UP 447 Varshney Anju 379
Nade VS 425 Woubshet Demissie 311

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Bacopa monnieri (L.) Wettst.,

Bacopa monnieri (L.) Wettst. locally known as water hyssop,

| IJLSCI Volume 5 | Issue 3|September 2017 | page nos.291- 479 http://www.ijlsci.in

425 Comparative study to evaluate in vivo and in LIST OF REFEREES

An Ethological and Ecological Review of the real-world

University of Chicago Laboratory Schools, 1362 E 59th St, Chicago, IL 60637,

Manuscript details: ABSTRACT

2017 |IJLSCI www.ijlsci.in 291

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Feeding ecology of tiger fish, (Hydrocynus vittatus) in the

Simasiku EK1*, Mafwila SK2, Abah J3 , Hay CJ 4, Matengu KK5

Bag 1096, Katima Mulilo, Namibia

*Corresponding author, e-mail: simasiku2000@yahoo.co.uk

Manuscript details: ABSTRACT

2017 |IJLSCI www.ijlsci.in 297

INTRODUCTION Zambezi River, highlighting the effects of fish size and

298 Int. J. of Life Sciences, Vol. 5(3) September, 2017

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RESULTS was observed on day 16 denoted by the gape in the

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Species Wet Dry

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Variation between seasons relationship. On average, H. vittatus consumed prey

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304 Int. J. of Life Sciences, Vol. 5(3) September, 2017

CONCLUSION Gilliam JF, Fraser DF (2001) Movement in corridors:

www.ijlsci.in Int. J. of Life Sciences, Vol. 5(3) September 2017 305

A Morphological Study of the Evolution of Eight Organisms

University of Chicago Laboratory Schools, 1362 E 59th St, Chicago, IL 60637,

Manuscript details: ABSTRACT

2017 |IJLSCI www.ijlsci.in 306

METHOD ancestor. In this paper, a mollusk is a group of animals

Table 1 Data Matrix

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Effect of integrated use of lime, blended fertilizer and compost on

Woubshet Demissie1, Selamyihun Kidanu2 and Cherukuri V Raghavaiah3*

1Soilfertility expert, Ministry of Agriculture, Addis Ababa, Ethiopia.

Manuscript details: ABSTRACT

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312 Int. J. of Life Sciences, Vol. 5(3) September, 2017

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Table 2. Chemical composition of compost used in the experiment.