Int. J. Curr. Sci. Tech. Vol. 1 No.

1, 2012

DEPLETION OF NUTRITIVE VALUES IN THE FLESH OF

FRESHWATER MUSSEL LAMELLIDENS MARGINALIS (LAMARCK)
AFTER DIMETHOATE EXPOSURE
Saurabh Kumar1, Rakesh Kumar Pandey2, Shobha Das3 and Vijai Krishna Das4
1
Department of Zoology, Ganpat Sahai P.G.College, Sultanpur (UP)-228001, India
ContacT No. 9415968343, e-mail; saurabhzw@gmail.com
2
Kamla Nehru Institute of Physical and Social Sciences, Sultanpur (UP) - 228118, India
Contact No. 9415968686, e-mail; rakeshzoology@gmail.com
3
Department of Zoology, Ganpat Sahai P.G.College, Sultanpur (UP)-228001, India
Contact No. 9918045600, e-mail; drshobhadas@gmail.com
4
Kamla Nehru Institute of Physical and Social Sciences, Sultanpur (UP) - 228118, India
Contact No. 9415074374, e-mail; dasvkster@yahoo.co.in

ABSTRACT : Contamination of freshwater bodies from the pesticides through surface run-off and sewage disposal are
well documented. Pesticides, which gain access into the water bodies cause unfavorable changes in the physico-chemical
characteristics of water and adversely affect not only the physiology and behavior of the economically important non-
target aquatic organisms but also alter the chemical composition of their flesh. To evaluate toxic effect of commonly
used pesticide dimethoate on chemical composition of flesh, the freshwater mussel Lamellidens marginalis were subjected
to short-term (75% concentration of 96h LC 50 for 24h, 48h, 72h and 96h) and long-term (25% concentration of 96h
LC 50 for 3d, 6d, 12d and 24 d) exposure. The results clearly indicate that dimethoate induces significant decrease in tissue
glycogen and protein contents (hepatopancreas and adductor muscle) of mussels, throughout the exposure.
Keywords : Pesticide, Dimethoate, Toxic effects, Nutritive value, Flesh, Mussel

INTRODUCTION Haryana, Jharkhand, Madhya Pradesh, Rajasthan,

Contamination of freshwater bodies from
the pesticides through surface run-off and sewage
disposal are well documented by several authors
[1, 2, 3, 4]. Pesticides, which gain access into the
water bodies cause unfavorable changes in the
physico-chemical characteristics of water and
adversely affect not only the physiology and
behavior of the economically important non-target
aquatic organisms but also alter the chemical
composition of their flesh. [5, 6, 7, 8, 9].
Freshwater mussel Lamellidens marginalis
(Unio) is a common bivalve occurring in
freshwater bodies like ponds, rivers, dams, ditches,
canals etc. in the ratio of 10-15 individual/m2 and
represented by 26 synonyms as well as 69 species
[10]. They are abundantly found in lower and
upper Gangetic plains of India, Bangladesh, Sri
Lanka, Myanmar and Terai region of Nepal. In
India, they are reported from Uttar Pradesh, Bihar,
Uttaranchal and West Bengal [10]. They are
ciliary-filter feeder, feeding mainly on planktons
(algae, protozoa and bacteria) as well as riverine
sediments. To obtain their food, they circulate
large quantity of water through their body cavity
from which the food substances are filtered out
with the help of cilia of gill filaments. They are
known to accumulate toxicants (Xenobiotics) like
heavy metals [11], pesticides [12, 13] etc. in their
body tissues, above the surrounding level, thus
enabling the identification as hot spot in the
ecosystem. Because of their detrivorous and
planktonic feeder habit, they accumulate heavy
metals and pesticides in their body therefore; they
are regarded as scavenger and purifier of the
ecosystem [14, 15].
The freshwater mussels are popularly
known for the production of pearl which has great
ornamental value. The shell is used to prepare

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 Saurabh Kumar, Rakesh Kumar Pandey, Shobha Das and Vijai Krishna Das
certain cheap jewelries and production of lime in
small scale industries. The mussel is used as food
by tribal people and poor families in India,
Bangladesh and Nepal [16]. Their flesh is rich in
protein, carbohydrate and lipid and is soft, tasty,
easily cooked and digestible (personal
communication with fishermen of Awadh region,
UP). More recently it has been observed that the
body extract of freshwater mussel has therapeutic
use in remedy of arthritis [17]. It also acts as
antimicrobial agent against different bacterial
strain [18].
Numerous biochemical indices of stress
have been proposed to assess the health of non-
target organisms exposed to toxic chemicals in
aquatic ecosystem (19, 1). Protein and
carbohydrate are important organic substances
required by organisms in tissue building and quick
energy production. They are intimately related
with almost all physiological processes, which
maintain simple biochemical system in living
condition. The freshwater mussel Lamellidens
marginalis is important food item in this area.
Hence, an attempt is made to study the changes
in total protein and glycogen content of
hepatopancreas and anterior adductor muscle as a
function of the effect of insecticide, dimethoate
during short and long-term exposure.
MATERIAL AND METHODS
Healthy freshwater mussel, Lamellidens
marginalis (Lamarck) were collected from river
Gomti and carefully brought to the laboratory
where, acclimatized for 10 days under laboratory
conditions in a 60 l capacity earthen tanks. The
crushed leaves of aquatic plants along with river
water were provided as food on alternate days.
Food was not given 24 h before the test. The
Physico-chemical characteristic such as dissolved
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oxygen (DO), temperature, pH, total hardness,
alkalinity (as CaCO 3) and free CO 2 of the test
solution were recorded daily following the
standard methods [20]. The experiments were
conducted during month of July-August 2010 at
room temperature (28.72.2 0C) with natural
photoperiod (12.51 : 11.090.31, light : dark). The
mussels of equal size (6.74 0.32 cm long and
3.51 0.32 cm wide) and weight (39.74 3.10
g) were selected.
We have determined 50% lethal
concentration of dimethoate on freshwater mussel
by using the standard method [20]. LC 50 values
were calculated from the data obtained in acute
toxicity tests by using EPA-Probit analysis version
1.5 statistical software, (http://www.epa.gov/
nerleerd/stat2.htm#tsk) based on Finneys method
[21]. The LC50 values obtained for dimethoate
were 45.09, 40.52, 38.71 and 36.35 mg/l for 24h,
48h, 72h and 96h respectively [22].
After determining LC50 of dimethoate, thirty
two acclimatized mussels were subjected to 75%
concentration of 96h LC50 of dimethoate (27.27
mg /l) for short term (24h, 48h, 72h, and 96h) and
25% concentration of 96h LC50 value (9.08 mg/l)
for long term (3d, 6d, 12d and 24d) respectively.
A control with equal number of mussel was also
run simultaneously. The experiment was carried
out in glass troughs of 15 l capacity in static
laboratory condition. Before starting the test all the
glass troughs were cleaned and filled with 14 l tap
water. The stock solution of 10mg/ml
concentration of dimethoate (Rogor, 30% EC,
Rallis India Ltd. Mumbai) was prepared by
dissolving in absolute alcohol. Eight mussels each
from treated and control of different time intervals
were removed from the troughs, wrapped dried
with tissue paper and dissected to remove tissue
samples. Tissue sample of hepatopancreas and
Int. J. Curr. Sci. Tech. Vol. 1 No. 1, 2012

anterior adductor muscle (200 mg each) were exposure and results are represented in the table
removed and grinded separately in 20 ml TCA 1.
(trichloroacetic acid) in a hand operated Table 1:- Physico-chemical characteristics of
homogenizer. The homogenate obtained was water used in the experiment.
processed according to method of van der Vies
S.No. Characteristics of water
[23] for the estimation of tissue glycogen and the
tissue protein was analyzed according to Lowery 1. Photoperiod 12.51:11.09
0.31, light:dark
et al. [24]. The results obtained were represented
as mg/100 mg wet tissue weight SD of eight 2. Dissolved oxygen 7.8 0.2 (mg/l)
mussels. 3. Free CO2 6.0 0.5 (mg/l)
The result of control and treated mussel at 4. pH 7.4 0.11
each time intervals were statistically analyzed for 5. Temperature (Air) 28.7 2.2C
the significance (P<0.05) and difference were 6. Temperature (Water) 28 0.5C
represented as % change, over the control. All the
7. Total Hardness 274 3.74 mg/l
statistical calculations are performed with the help
of Microsoft Excel Data analysis Programme 8. Alkalinity (as CaCO3) 180 4.50 mg/l
and graphpad (http://www.graphpad.com/ The mussels exposed to dimethoate for short
quickcalcs/ttest1.cfm?Format=SD) online and long term show gradual decline in glycogen
calculator. and total protein levels in the tissue of
hepatopancreas and anterior adductor muscle of
RESULTS
both control as well as treated mussels. However,
The physico-chemical characteristics of test decrease in treated mussel is rapid and represented
water such as temperature (air and test water), DO, statistically significant decrease over control
free CO2, alkalinity, total hardness and pH which (Table 2 and 3).
were regularly monitored throughout the period of

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 Saurabh Kumar, Rakesh Kumar Pandey, Shobha Das and Vijai Krishna Das

Table 2: Glycogen, in the hepatopancreas and adductor muscle of Lamellidens marginalis and % change in
comparison to control when exposed for short (96h duration) and long (24 Days) term duration to sub lethal
concentration of dimethoate. Values are MeanSD of eight animals. Asterisk indicate insignificant (*) and
significant (**) differences (P<0.05) in percent change over control.
Duration Duration Hepatopancreas tissue glycogen Adductor muscle tissue glycogen
(Hours) (Hours) (mg/100mg, wet tissue weight) (mg/100mg, wet tissue weight)
24h 48h 72h 96h 24h 48h 72h 96h
Short term Control 4.88 4.77 4.64 4.03 4.25 4.11 4.05 3.90
Exposure (96 h) 0.20 0.14 0.45 0.39 0.31 0.08 0.08 0.67
Treated 3.48 3.21 3.03 2.69 3.98 3.88 3.48 3.27
0.36 0.45 0.10 0.53 0.12 0.17 0.64 0.48
% change 28.69** 32.70** 34.70** 33.25** 6.35** 5.60** 14.07** 16.15**
over control
Duration 3d 6d 12d 24d 3d 6d 12d 24d
Long term Control 4.72 4.57 4.42 4.22 3.85 3.81 3.72 3.52
Exposure (24 d) 0.35 0.29 0.39 0.27 0.11 0.47 0.48 0.57
Treated 3.22 3.02 2.90 3.70 3.81 3.79 3.61 2.79
0.26 0.13 0.06 0.25 0.69 0.68 0.10 0.11
% change 35.12** 33.91** 34.39** 12.32** 1.03* 0.52* 3.37* 20.73**
over control

Table 3: Total protein, in the hepatopancreas and adductor muscle of Lamellidens marginalis and % change in
comparison to control when exposed for short (96h duration) and long (24 Days) term to sub lethal concentration
of dimethoate. Values are MeanSD of eight animals. Asterisk indicate insignificant (*) and significant (**)
differences (P<0.05) between control and treated.
Duration Duration Hepatopancreas tissue protein Adductor muscle tissue protein
(Hours) (Hours) (mg/100mg, wet tissue weight) (mg/100mg, wet tissue weight)
24h 48h 72h 96h 24h 48h 72h 96h
Short term Control 28.00 27.12 26.00 25.12 31.06 29.81 27.90 25.85
Exposure (96 h) 1.69 2.85 1.69 3.09 0.82 0.98 0.69 0.79
Treated 27.00 20.25 20.00 18.00 29.93 22.30 20.05 15.59
1.69 1.98 2.00 1.92 0.56 0.76 0.73 0.89
% change 3.58** 25.33** 23.08** 28.34** 3.63** 25.20** 28.13** 39.70**
over control
Duration 3d 6d 12d 24d 3d 6d 12d 24d
Long term Control 23.00 22.00 19.00 18.00 22.00 21.00 18.00 17.00
Exposure (24 d) 1.19 2.32 1.77 2.00 1.41 1.77 1.69 1.30
Treated 16.00 15.12 12.00 10.88 21.00 15.88 11.70 6.25
1.41 1.24 1.30 1.35 2.39 1.24 1.12 1.03
% change 30.43** 31.28** 36.84** 39.55** 4.54* 24.3** 35.00** 63.23**
over control

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Int. J. Curr. Sci. Tech.
The decrease in tissue protein and glycogen
of mussels exposed for long term is greater than
mussels exposed for short term. The mussels
exposed for long term show 39.55% decline in
hepatopancreas and 63.23% decline in adductor
muscle tissue protein while the reduction was only
28.34% in hepatopancreas and 39.70% in adductor
muscle during the short term exposure. Similarly,
tissue glycogen of hepatopancreas and adductor
muscle was declined up to 35.39% and 20.73%
respectively during long term exposure while it
was only 34.70% and 16.15% respectively during
the short term exposure (Table 2 and 3). The
glycogen level of hepatopancreas in exposed
mussels is more sensitive to dimethoate toxicity
and shows more rapid decline in comparison to
adductor muscle. However, following 24d
exposure the decline in glycogen level of the
adductor muscles was greater than the
hepatopancreas (Fig. 1). In contrast, the tissue
protein level of hepatopancreas depleted slowly in
comparison to the adductor muscles through out
the exposure (fig. 2).
DISCUSSION
Freshwater mussel Lamellidens marginalis
is known to have high percentage of protein and
glycogen in their flesh (16, 25). Protein is the chief
and carbohydrate is the second biochemical
constituent of marine bivalve (oyster) Crassostrea
madrasensis and Perna viridis [25].
Hepatopancreas (digestive gland) is the metabolic
reservoir, which easily mobilizes the stored food
materials viz. glycogen, to meet out the sudden
energy requirements under physiological stress
created due to pesticide [26] and heavy metal [27]
exposures. Hepatopancreas (digestive gland) of
bivalve molluscs is the principal site of
intracellular digestion, absorption and metabolic
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Vol. 1 No. 1, 2012
detoxification of various organic and inorganic
toxic substances [28, 29, 30] and known to possess
high level of stress protein glutathione-s-
transferase (GST) which has its role in
detoxification and removal of xenobiotics. The
GSTs is recognized as a biomarker of pollution
and stress [31, 32]
The toxicity of pesticides and heavy metals
to the bivalve molluscs has been studied by several
workers (33, 34, 35, 36, 37). Pesticide exposure
leads to decline in glycogen and total protein
levels in the tissues of bivalves (33, 35, 36).
The initial decrease in the hepatopancreas
glycogen of Lamellidens after dimethoate
exposure is due to immediate mobilization of
glycogen as glucose for energy production. This
clearly indicates that mobilization of glycogen first
takes place from the hepatopancreas followed by
the muscles. Protein is a structural component of
animal body; under stressful conditions it may be
mobilized through gluconeogenesis to compensate
the energy demand after the glycogen stores of
body is depleted followed by lipids. In present
study muscles and hepatopancreas exhibit a
reduction in total protein levels after 48 to 96h and
a progressive decrease in long term exposure up
to 24d.
The present work clearly indicates that
dimethoate pesticides which are used vigorously
in the houses and agricultural practices, if get entry
into the freshwater bodies, produces adverse
effects on inhabiting non-target organisms. So, this
pesticide should be sold in the market with
cautions on the packets and bottles and using safe
procedures of application.
ACKNOWLEDGEMENT
The authors are thankful to Dr. P. B. Singh,
Principal (G. S. P. G. College) and Dr. Gunraj
 Saurabh Kumar, Rakesh Kumar Pandey, Shobha Das and Vijai Krishna Das

Prasad, Principal (KNIPSS) for providing research 320.

18. M. Ester, J. Satyanarayana, B. S. Kumar, T. Bikshapati,
facilities during the course of our research work. A. S. Reddy, L. Venkanna, Biol. Med. 3 (2011) 191-
195
REFERENCES 19. A. J. Nimmi. J. Great Lakes Res. 16 (1990) 529-541
1. W. E. Pereira, J. L. Domagalski, F. D. Hostettler, L. R. 20. APHA, Standard methods for the examination of water
Brown, J. B. Rapp, Environ. Toxicol. Chem. 15 (1996) and waste water. 20 th edn. APHA, AWWA, WPCF,
172-180. Washington DC, USA, 2005.
2. H. Toshiro, S. Ytaka, Ins. Hyg. Environ. 29 (2004). 21. D. J. Finney, Probit analysis. University Press
3. L. Kiaune, N. Singhasemanon, Reviews of, Environ. Cambridge, 1971.
Contam. Toxicol. 213 (2011), DOI :-10.1007/978-1- 22. S. Kumar, R. K. Pandey, S. Das and V. K. Das, Res.
4419-9860-6_1, Res. Sci. Tech. (2011) (Under communication)
4. B. Kumar, R. Gaur, G. Goel, M. Mishra, D. Prakash, S. 23. J. Van der Vies, Biochem. J. 57 (1954) 410-416
K. Singh, R. B. Lal, S.Kumar, C. S. Sharma, Asian J. 24. O. H. Lowry, N. J. Rosebrough, A. L. Farr, R. J. Randall,
Scie. Res. 4 (2011) 271-280 J. Biol. Chem 193(1951) 265 (The original method).
5. C. A. Moulten, W. J. Fleming, C. E. Purnell, Environ. 25. G. M. Salaskar, V. N. Nayak, Rec. Res. Scie. Tech. 3
Toxicol. Chem. 15 (1996) 131-137. (2011) 06-11.
6. A. E. Keller, D. S. Ruessler, Environ. Toxicol. Chem. 26. N. S. EI-shenawy, T. I. S. Moawad, M. E. Mohallal, I.
16 (1997) 1028-1033. M. Abdel-Nabi, I. A. Taha . Ocean Scie. J. 44
7. C. S. Kontreczky, A. Farkas, J. Nemesok, J. Salanki, (2009) 27-34.
Ecotoxicol. Environ. Saf. 38 (1997) 195-199. 27. A. V. Andhale, P. A. Bhosale, S. P. Zambare, J. Exp.
8. S. Das, B. B. Jana, Ind. J. Exp. Biol. 41 (2003) 1306- Sci. 2 (2011) 01-03
1310. 28. P. S. Rainbow, D. J. H. Phillips, Mar. Poll. Bull. 26
9. N. Arab, C. Minier, S. Lemaire, E. Unruh, P. D. Hansen, (1993) 593-603.
B.K. Larsen, O. K. Anderson and J.F. Narbone, Mar. 29. I. Marigomez, M. Soto, M. P. Cajaraville, E. Angulo,
Environ. Res. 58 (2004) 437-441. L. Giamberini, Microse Res. Tech. 56 (2002) 358-392.
10. A. Biswas, S. K. Raut, J. Nat. Tai. Mues. 52 (1999) 78- 30. L. N. Usheva, M. A. Vaschenko, V. B. Durkina, Russian
85. J. Mar. Biol. 32 (2006) 166-172.
11. H. M. Santosh, M. S. Diniz, P. M. Costa, I. Peres, M. 31. B. N. Blnchette, B. R. Singh, J. Protein Chem. 21(2002)
H. Costa, S. Alves, J. L. Capelo, Wiley Periodicals, Inc. 489-494.
Environ. Toxicol. 22 (2007) 502509.
32. S. J. Won, A. Novillo, N. Custodia, M.T. Rie, K.
12. S. Uno, H. Shiraishi, S. Hatakeyama, A. Otsuki, J. Fitzgeald, M. Osada, I. P. Callard, Integr.
Koyama, Arch. Environ. Contam. Toxicol. 40 (2001) Comp. Biol. 45 (2005) 72-80.
35-47.
33. K. S. Moorthy, M. D. Naidu, C. S. Chetty, K. S. Swamy,
13. A. K. Chopara, M. K. S. Sharma and S. Chamoli, Bull. Environ. Contom. Toxicol. 30 (1983) 219-222.
Environ. Monit. Assess.173 (2011) 905-916.
34. K. Satyaparameshwar, T. R. Redy, N. V. Kumar, J.
14. K. Simkiss, M.Taylor, A. Z. Mason, Mar. Biol. Lett. 3. Environ. Biol. 27 (2006) 39-41.
(1982) 187-201.
35. B. Gagnaire, M. Gay, A. Huvet, J.Y. Daniel, D. Saulnier,
15. D. R. Livingstone, Comp. Biochem. Physiol. (c) 100 T. Renault, Aqua. Toxicol. 84 (2007) 92-102.
(1991) 151-155.
36. N. S. EI-shenawy, R. Greenwood, I. M. Abdel-Nabi,
16. R. L. Baby, I. Hasan, K. A. Kabir, M. N. Naser, J. Scie. Acta Zoologica Sinica 53 (2007) 899-909.
Res. 2 (2010) 390-396.
37. F. Dondero. M. Banni, A. Negri, L. Boatti, A. Dagnino
17. M. Cakraborthy, S. Bhattacharya, P. Bhattacharjee, R. and A. Viarengo, BMC Genomics 12 (2011)
Das, R. Mishra, J. Ethonophormacol 132 (2010) 316- 195 (http://www.biomedcentral.com/1471-2164/12/195)

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Int. J. Curr. Sci. Tech. Vol. 1 No. 1, 2012

Fig. 1: % change in quantity of tissue glycogen of
freshwater mussel Lamellidens marginalis over
control in different periods of exposure. Values are
MeanSD of eight animals. Asterisk indicate
insignificant (*) and significant (**) differences
(P<0.05) between control and treated mussels.
Fig. 2: % change in quantity of tissue protein of
Lamellidens marginalis over control in different
periods of dimethoate exposure. Values are
MeanSD of eight animals. Asterisk indicate
insignificant (*) and significant (**) differences
(P<0.05) between control and treated mussels.

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