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PHYTOCHEMICAL ANALYSIS OF HALODULE UNINERVIS

IN PANGIL CURRIMAO ILOCOS NORTE

MARK GIL GELBOLINGO

COLLEGE OF AQUATIC SCIENCES AND APPLIED TECHNOLOGY

MARIANO MARCOS STATE UNIVERSITY

CURRIMAO, ILOCOS NORTE

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE BACHELOR


OF SCIENCE IN MARINE BIOLOGY
PHYTOCHEMICAL ANALYSIS OF HALODULE UNINERVIS

IN PANGIL CURRIMAO ILOCOS NORTE

INTRODUCTION

Background of the study

This species is a flowering plant spreading via a branching rhizome that roots
at orne on a short peduncle and is enclosed in a leaf. The tiny anthers are red. The
fruit is about 2 millimeters long.

Leaf morphology changes according to habitat type. The leaves are wider in
deeper waters. There are apparently two morphs, a narrow leaf and a wide leaf,
rather than a continuous range. The narrow leaf type is found closer to shore where
it is exposed more often. The wide leaf type is found in deeper areas with cloudier
waters. Plants that receive less light may need more leaf blade area to perform
enough photosynthesis.

This grass forms dense carpets or meadows on the substrate, sometimes


mixing with other seagrasses and algaes.It occupies the lower intertidal zone, and it
is less tolerant of exposure to air than are plants of the upper intertidal zone such as
Thalassia hemprichii. It desiccates quickly. It is also sensitive to ultraviolet
radiation. These factors restrict it to deeper intertidal waters than some other
plants.It is a euryhaline species, tolerating a wide salinity range.

This species is an important food for the dugong. The grass grows in the
Masirah Channel, a waterway between Masirah Island and mainland Oman, where
it is an important food for the green sea turtle.
This is a pioneer species. It has been observed on high-sediment, rapidly
evolving substrates in Australia and Indonesia.This species is known to be
hybridized to Halodule pinifolia in Okinawa, Japan.

This plant is widely distributed and it is common throughout its range. In


general its populations are stable, though it may be decreasing in localized areas,
such as the coast of Bangladesh, and it fluctuates in some Australian waters. It is
affected by some degradation of habitat by forces such as coastal development,
siltation, sedimentation, weather events and tidal action, predation, parasites,
disease, trawling and other fishing practices, dredging, pollution, eutrophication,
and climate change.

Conservation plans are in effect in various regions. Populations are


monitored in the United Arab Emirates. It grows within the bounds of several
marine parks and reserves in Africa. Populations can be disturbed only with
permits in parts of Australia. Large beds are protected in Hat Chao Mai National
Park in Thailand. This is a common plant of the sublittoral zone in its range,
growing in depths up to 20 meters in lagoons, on reefs, and in many other types of
marine habitat just offshore. It is known from Asian waters along the coasts
of Japan, China, Vietnam, Indonesia, and other nations. It occurs on Pacific
Islands such as Fiji. It occurs along the Australian Pacific coast, including
the Great Barrier Reef. It can be found along Indian Ocean coastal regions from
Australia to India to eastern Africa. It is resident in the Red Sea and Persian Gulf

It produces erect stems and alternately arranged leaves. The narrow, toothed leaf
blades are up to 15 centimeters long and usually roughly a millimeter wide, though
leaf width is variable and can be up to 7 millimeters. Each leaf has a sturdy sheath
up to 3.5 centimeters long. The tip of the leaf blade has three teeth. Plants of this
family are dioecious. The male flower is borne on a short peduncle and is enclosed
in a leaf. The tiny anthers are red. The fruit is about 2 millimeters long.
Seagrasses are the marine flowering plants. Seagrasses such as Amphibolis,
Heterzostera, Phyllospadi, Posidonia, Pseudalthenia and Zostera are mostly
restricted to temperate seas and seven genera of seagrasses such as Cymodocea,
Enhalus, Halodule, Halophila, Syringodium, Thalasia and Thalassodendeon are
distributed in tropical seas. Sea grass biomass is used as human food especially by
coastal populations. Numerous seagrasses have been shown to have antibacterial
activities. Halophila stipulacea, Cymodocea serrulata and Halodule pinifolia,
Enhalus acoroides and Enhalus acoroides, Thalassia
hemprichii, Halodule pinifolia, Syringodium isoetifolium, and Cymodocea
rotundata have been reported to exhibit antibacterial activity. Moreover,
preliminary data suggest that seagrasesses could represent an interesting source of
antilarvacidal and antioxidant. However, there is a little information regarding
seagress from Halodule uninervis and their antioxidative activity This is a common
plant of the sublittoral zone in its range, growing in depths up to 20 meters
in lagoons, on reefs, and in many other types of marine habitat just offshore. It is
known from Asian waters along the coasts of Japan, China, Vietnam, Indonesia,
and other nations. It occurs on Pacific Islands such as Fiji. It occurs along
the Australian Pacific coast, including the Great Barrier Reef. It can be found along
Indian Ocean coastal regions from Australia to India to eastern Africa. It is resident
in the Red Sea and Persian Gulf. This plant is widely distributed and it is common
throughout its range. In general its populations are stable, though it may be
decreasing in localized areas, such as the coast of Bangladesh, and it fluctuates in
some Australian waters. It is affected by some degradation of habitat by forces
such as coastal development, siltation, sedimentation, weather events and tidal
action, predation, parasites, disease, trawling and other fishing
practices, dredging, pollution, eutrophication, and climate change. Conservation
plans are in effect in various regions. Populations are monitored in the United Arab
Emirates. It grows within the bounds of several marine parks and reserves in
Africa. Populations can be disturbed only with permits in parts of Australia. Large
beds are protected in Hat Chao Mai National Park in Thailand

Objectives of the study


This study was to determine the following objectives:

1. Study the seagrass halodule uninervis if it is posible to make it a medicine.

2. To observe the effect of changing temperature in seagrass.

3. To observe the effect of medicinal seagrass in human body.

Scope and limitations

The study was limited to seagrass organism that was subjected to the
phytochemical analysis. The experiment is to Study the seagrass halodule uninervis
if it is posible to make it a medicine, To observe the effect of changing
temperature in seagrass and to observe the effect of medicinal seagrass in human
body.
REVIEW OF RELATED LITERATURE

Biology of halodule uninervis

Figure 1. Photo of halodule uninervis

Kingdom: Plantae

Clade: Angiosperms

Clade: Monocots

Order: Alismatales

Family: Cymodoceaceae

Genus: Halodule

Species: H. uninervis

Scientific name: Halodule uninervis


Taxonomy, Halodule uninervis has a wide Indo-Pacific distribution. In the Pacific,
it is found in southern Japan,Taiwan, Philippines, Malaysia, Indonesia, throughout
the Gulf of Thailand and along the coast of Vietnam and southern China. It occurs
throughout insular Southeast Asia, northeast to the Northern Mariana Islands,
Micronesia, and southeast to the Fiji Islands, as well as across northern Australia
and the Great Barrier Reef. This grass forms dense carpets or meadows on the
substrate, sometimes mixing with other seagrasses and algaes.
It occupies the lower intertidal zone, and it is less tolerant of exposure to air than
are plants of the upper intertidal zone such as Thalassia hemprichii. It desiccates
quickly. It is also sensitive to ultraviolet radiation. These factors restrict it to deeper
intertidal waters than some other plants. It is a euryhaline species, tolerating a
wide salinity range. This grass forms dense carpets or meadows on the substrate,
sometimes mixing with other seagrasses and algaes. It occupies the lower intertidal
zone, and it is less tolerant of exposure to air than are plants of the upper intertidal
zone such as Thalassia hemprichii. It desiccates quickly. It is also sensitive
to ultraviolet radiation. These factors restrict it to deeper intertidal waters than
some other plants. It is a euryhaline species, tolerating a wide salinity range

Morphology
The wide range of leaf types that occurs within the populations
of Halophilaand Halodule in Shark Bay, Western Australia, was studied for plants
under laboratory conditions. In the Halophila ovalis-H. minor complex, two types
of plants appeared in the cultures, a large-leaved variant that is similar to typical H.
ovalis (R.Br.) Hook. f. and a small-leaved variant that is more similar to H.
minor (Zoll.) den Hartog. In Shark Bay, the two variants were not clearly separated
because both produced leaves that varied in size, degree of waviness of leaf
margins and intensity of anthocyanin pigmentation. In the Halodule
uninervis (Forsk.) Aschers. complex, two types of plants, wide- and narrow-leaved
variants, appeared in the cultures. In Shark Bay, the narrow-leaved plants differed
in leaf width in muddy and silty microsites, but were always narrower than the
wide-leaved variants. Only the narrow-leaved Haloduleplants produced
anthocyanin pigmentation both under field and laboratory conditions. The studies
under controlled conditions indicate that the highly diverse leaves in Shark Bay
result primarily from environmental influences on two types of Halophila and two
types of Halodule.
Ecology

This species is an important food for the dugong. The grass grows in the Masirah
Channel, a waterway between Masirah Island and mainland Oman, where it is an
important food for the green sea turtle. This is a pioneer species. It has been
observed on high-sediment, rapidly evolving substrates in Australia and Indonesia.
This species is known to be hybridized to Halodule pinifolia in Okinawa, Japan.

Importance

Seagrasses are submerged flowering plants found in shallow marine waters,


such as bays and lagoons and along the continental shelf in the Gulf of Mexico. A
vital part of the marine ecosystem due to their productivity level, seagrasses
provide food, habitat, and nursery areas for numerous vertebrate and invertebrate
species. The vast biodiversity and sensitivity to changes in water quality inherent
in seagrass communities makes seagrasses an important species to help determine
the overall health of coastal ecosystems. Seagrasses perform numerous functions:

 Stabilizing the sea bottom


 Providing food and habitat for other marine organisms
 Maintaining water quality
 Supporting local economies

Stabilization: Ocean bottom areas that are devoid of seagrass are vulnerable to
intense wave action from currents and storms. The extensive root system (see
diagram below) in seagrasses, which extends both vertically and horizontally, helps
stabilize the sea bottom in a manner similar to the way land grasses prevent soil
erosion. With no seagrasses to diminish the force of the currents along the bottom,
Florida's beaches, businesses, and homes can be subject to greater damage from
storms.

Ecosystem support: Seagrasses provide food, shelter, and essential nursery areas
to commercial and recreational fishery species and to countless invertebrates living
in seagrass communities. Some fish, such as seahorses and lizardfish, can be found
in seagrasses throughout the year, while other fish remain in seagrass beds during
certain life stages.

Food: While some organisms, including the endangered Florida manatee and green
sea turtle, graze directly on seagrass leaves, others use seagrasses indirectly to
provide nutrients. Bottlenose dolphins are often found feeding on organisms that
live in seagrass areas. Detritus from bacterial decomposition of dead seagrass
plants provides food for worms, sea cucumbers, crabs, and filter feeders such as
anemones and ascidians. Further decomposition releases nutrients (such as
nitrogen and phosphorus), which, when dissolved in water, are re-absorbed by
seagrasses and phytoplankton.

Nursery areas: The relative safety of seagrass meadows provides an ideal


environment for juvenile fish and invertebrates to conceal themselves from
predators. Seagrass leaves are also ideal for the attachment of larvae and eggs,
including those of the sea squirt and mollusk. Much of Florida's recreationally and
commercially important marine life can be found in seagrass meadows during at
least one early life stage.

Habitat: While seagrasses are ideal for juvenile and small adult fish for escape
from larger predators, many infaunal organisms (animals living in soft sea bottom
sediments) also live within seagrass meadows. Species such as clams, worms,
crabs, and echinoderms, like starfishes, sea cucumbers, and sea urchins, use the
buffering capabilities of seagrasses to provide a refuge from strong currents. The
dense network of roots established by seagrasses also helps deter predators from
digging through the substratum to find infaunal prey organisms. Seagrass leaves
provide a place of anchor for seaweeds and for filter-feeding animals like
bryozoans, sponges, and forams.

Water Quality: Seagrasses help trap fine sediments and particles that are
suspended in the water column, which increases water clarity. When a sea floor
area lacks seagrass communities, the sediments are more frequently stirred by wind
and waves, decreasing water clarity, affecting marine animal behavior, and
generally decreasing the recreational quality of coastal areas. Seagrasses also work
to filter nutrients that come from land-based industrial discharge and stormwater
runoff before these nutrients are washed out to sea and to other sensitive habitats
such as coral reefs.
Economics: Although seagrass is not a commodity that is directly cultivated in
Florida, its economic value can be measured through other industries, such as
commercial and recreational fisheries and nature and wildlife tourism, which rely
on this habitat to survive. Since most of Florida's fishery species (approximately
70%) spend at least part of their life cycle within seagrass communities, seagrasses
are vital to the survival of these fishing industries.
METHODOLOGY

Preparation of Sample

Sea grass of Halodule uninervis was collected from Coastal of pangil and
immediately brought to the laboratory in sterile plastic bags containing water to
prevent evaporation. Seagrasses were washed thoroughly with water to remove
extraneous materials and shade-dried for 10 days at room temperature until
constant weight obtained. The dried Seagrasses were powdered and stored in
refrigerator for future use.

Preparation of Sea grass extract

Seagrass powder were soaked in 2 L with methanol (1:4 w/v), and kept for
10 days in a shaker. The extraction was repeated thrice and pooled. The dry
aqueous extracts were stored in a refrigerator until further analysis.

Phytochemical Screening of Halodule uninervis

Flavonoids Test of flavonoids were determined by Wadood et al method.


A sample was heated with 10 ml of ethylacetate over a steam bath for 3 min. The
mixture was filtered and 4 ml of the filtrate was shaken with 1ml of dilute
ammonia solution. A yellow colouration indicated the presence of flavonoid.
Alkaloids Test of alkaloids were determined by Trease & Evan method. Saponins
Test of saponin were determined by Wall et al method [10]. A sample was shaken
with water in a test tube. Frothing which persists on warning confirmed the
presence of saponins. Steroids Test of steroids were determined by Edeoga et al
method. 1 ml of acetic anhydride was added twice to 0.5 g of aqueous extract with
2 ml H2S04. The colour changed from violet to blue or green in sample indicating
the presence of steroids
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