IJBI 1 (2), (DECEMBER 2019) 73-77

International Journal of Biological Innovations

Available online: http://ijbi.org.in | http://www.gesa.org.in/journals.php

DOI: https://doi.org/10.46505/IJBI.2019.1207

Research Article E-ISSN: 2582-1032

Cadmium induced histopathological alterations in female gonad

of freshwater bivalve mollusks, Lamellidens marginalis
during summer season

Shaikh Yasmeen

Department of Zoology
Dr. Rafiq Zakaria College for Women, Aurangabad (M.S.) India
*Corresponding author: [email protected]

Received: 20.11.2019 Reviewed: 02.12.2019 Accepted: 15.12.2019

Abstract: The study was aimed to investigate the effect of cadmium on the gonad in female fresh water bivalve mollusks,
Lamellidens marginalis. The experiment was carried out on the bivalves collected from Kutluq Lake, Daultabad, district
Aurangabad of Maharashtra, India from January 2018 to December 2018. The histolopathological findings include partial
disruption of ovarian follicle, vacuolation of germinal cells cytoplas m and damaged inter follicular connective tissue. The
cytomorphological structure of ovarian follicles got deformed and elongated, losing their typical configuration. Necrosis and
fibrosis in the connective tissue and damage to yolk vesicles of maturing sites were observed.

Keywords: Cadmium, Female gonad, Histopathology, Lamellidens marginalis.

INTRODUCTION In order to understand a pattern of damage caused by

Histopathology is an indispensable and powerful technique particular chemical to the tissue, it is essential to have an
in establishing routine toxicology studies performed for the insight into the histological analysis of the tissues.
purpose of risk assessment of living resources useful Histopathology, deal with the study of pathological changes
exposing the tissue contents and mechanisms of action at of the microscopic structure of the body tissue. Any peculiar
cellular level. To assess its usefulness in toxicology studies type of alteration of cells may indicate the presence of the
with shellfishes, several investigators have performed disease or the effect of toxic substances. Every organism has
experiments using various environmental contaminants capacity to tolerate suboptimal stress conditions. The
(Wester and Canton, 1991). maximum tolerance is at the extreme stress condition which
exhibits physiologically defined limitations, reflected
Many species of bivalve mollusks abundantly found in Indian predominantly in the structural architecture of various tissues
waters can sustain regular & very productive Fisheries in of the animal. Traces of toxicants introduced in the body can
India, particularly in Maharashtra state. Several species of be neutralized by immune system, but when high amount of
commercial, important and edible bivalves like clams, toxicant enters, it affects the structure and function of
oysters mussels etc. are found along the coastal areas whereas different organs in the body of animal. Thus, histopathology
other bivalve shell fishes like mussels and clams are found is an extremely useful tool for assessing effects of toxicants at
individual level. Their uncontrolled use may cause profound
both in lotic and lentic freshwater bodies. Both marine and
effects and a long term environmental impact on natural
freshwater bivalve shellfishes play an important role as bio-
aquatic environments (Bellas et. al., 2004). Pesticides may
indicators to detect various environmental fluctuations impair the functioning of organisms in aquatic environment
(Dholakia 2013 and Shinde Nitin 2017). even at low concentration.
74 International Journal of Biological Innovations 1 (2), (DEC. 2019)

Histopathological abnormalities caused due to toxicity of In the light of above researches, the present investigation was
pesticides have been reported earlier by many investigators undertaken with a view to study the histopathological
(Muley and Mane, 1990; Jonnalagadda and Rao, 1996; changes in the female gonad of Lamellidens marginalis,
Waykar, 1998) Zhou et. al., (1993) studied the histological under cadmium toxicity.
changes in the ovotestes of snail Biomphalaria glabrata on
exposure to molluscicides. Jyothi and Narayan (1996) MATERIALS AND METHODS
studied the effect of organophosphorous insecticide phorate After collection of the animals from habitat, they were
on gonads of freshwater fish Clarius batrachus. Kumari and immediately transported to the laboratory. The fouling and
Kumar (1997) studied the histopathological alterations in mud on shell valves were removed without disturbing the
ovary induced by aquatic pollutants in Channa punctatus. siphonal regions. The equal sized animals (90-100 mm shell
Hazarika and Das (1998) investigated the histopathological length) were grouped and kept in sufficient quantity of water
changes induced in ovary of Heteropneustes fossilis exposed (animal/liter) in aquaria with aeration for 24 hrs to adjust the
to BHC. Khan and Jha (2000) studied gonad histopathology animals in laboratory conditions (with renewal of water at
of the freshwater fish, Channa punctatus after exposure to interval of 12 to 13 hrs). No food was given during
phosalone. Ramchandra Mohan (2000) studied malathion acclimation time and during experiments. After 24 hrs, 05
induced changes in the ovary of freshwater fish, Glossobius groups of animals of almost equal size (90-100 mm shell
quaris. Baruah and Das (2002) studied histopathological length) were formed and each group with 10 animals
changes in ovary of fish Heteropneustes fossilis exposed to including control group and exposed to different test
paper mill effluent. Phirke (2008) studied the effect of concentrations of cadmium for static bioassay tests. The
quinolphos and thiodan on gonads of freshwater bivalve, stock solution of cadmium was prepared by was made
Parreysia corugata. Otitoloju et. al., (2009) studied the dissolving appropriate quantity of cadmium chloride (CdCl2
histopathlogy and bioaccumulation of heavy metals in 2½H2O AR Grade CDH Bombay) in double distilled water.
ovotestes of giant land snail, Archachatina marginata. The pH of the water is brought between 6.9 and 7.1 by adding
Kandasamy and Muthukumaravel (2010) studied the toxic 1N HCl (due to insolubility of cadmium in reservoir water
effect of chromium on the histoarchitectural alterations in having 7.6 to 8.1).
ovary of freshwater fish, Oreochromis mossambicus.

Photograph showing: I from North to South and II. From West to East

Appropriate test concentrations were then prepared and RESULTS AND DISCUSSION
animals were exposed. The experiments were conducted in The polluting substance could affect the histology of the
natural day-night rhythm. The experiments were repeated organism, meaning the deterioration of tissue and damage at
three times for confirming observed LC0 and LC50 values. The the cellular level. Severe damage at the cellular level was
96 hrs acute test was recorded. After 96 hrs acute toxicity test observed in female gonad after acute exposure. As compared
using cadmium for histological studies, gonad from control, to tissues of a control group of bivalve mollusks, these
LC0 and LC50 groups were fixed in aqueous Bouin's Hollande histological changes were prominent after acute exposure as
fluid for 48 hrs. The dissected tissues were dehydrated with compared to control group.
alcohol and toluene and embedded in paraffin wax (58 to
During the experimental period, the female gonad showed a
60°C). Gonad sections were cut down at 6-7µ. For
developmental condition of gonad. In the control group,
histopathological study of gonad Mallory's triple stain was
female follicle showed follicle wall with prominent germ
used. All the photomicrography was made under light-
cells and vitellogenic oocytes of different sizes. Many free
microscope Labo VT-20 Scan model.
Shaikh Yasmeen 75

mature eggs possessed dense cytoplasm and distinct nucleus vitellogenic oocytes lost their shape and detached from
located in centre having prominent nucleolus. The staining of follicle wall. Swelling of follicle was considerable (Fig. 2).
these different parts in female follicle was distinct (Fig.1).
After acute exposure to cadmium LC 0 group and LC 50 group In LC50 group, the follicle wall was distorted, mostly showed
of bivalves showed considerable damage to the female gonad prominent nuclei, nucleoli and the cytoplasm was opaque.
as compared to control group. In LC0 group, the follicle wall These are likely to undergo degeneration. Small sized
ruptured at places with shrinkage of germ cells along the wall. previtellogenic oocytes also showed such conditions. Nature
Deterioration of ooplasmic material, the nucleus and nucleoli of damage was more severe in LC50 than the LC0 group (Fig.
were observed in mature eggs. The germ cells and 3).

1. Control group 2. LC0 group 3. LC50 group

T. S. of Female gonad of Lamellidens marginalis

Fig. 1: Control group. Fig. 2:LC0 group. Fig. 3: LC50 group
F: Follicle MG: Mature Gamete, GO: Growing Oocyte, GC: Germ Cell, C: Connective Tissue, N: Nucleus,
DF: Distorted Follicle, LST: Loss of Stroma, ST: Stroma

Histological changes in gonads: (Fig 1to3): biochemical disturbances in aquatic organisms like mollusc
due to pesticide toxicity are well documented, Waykar and
Control Lomte (2002 and 2004). Histopathological changes are
In female ovaries many vitellogenic oocytes were found and mostly confined to organs directly involved in their
being released. In few follicles degenerating oocytes were metabolism and detoxification, Rashatwar and Ilyas (1994).
also seen. In the control group, during maturation the lumen of a female
follicle was filled with developing oogonia and primary
LC0 and LC50 groups
oocytes. Few follicles showed primary germ cells along the
In summer, LC0 group showed the formation of many pre
wall. The nuclei of the oocytes and ripened gametes were
vitellogenic and vitellogenic oocyte. Prominent vacuole conspicuous, but chronic stress from experimental group
appeared in their nuclei and lipid globules and nutritive cells showed rupture of follicle at some places, discontinuous
decreased much in quantity. In LC50 group, follicles distorted germ cells, loss of continuity in the follicle wall, dislocation
and ooplasmic material and nuclei showed vacuoles, of nucleus, internal follicular edema in ovary and disrupted
vitellogenic oocyte were also formed. Prominent vacuole basement membrane. Comparing the effects of Cadmium on
appeared in their nuclei and lipid globules and nutritive cells female gonads of bivalves, it was observed that, the effects
decreased much in quantity. In LC50 group, follicles distorted were more prominent in LC50 group, followed by LC0 group.
and ooplasmic material and nuclei showed vacuoles. Follicular shrinkage and distortion was observed in LC0, LC50
and chronic groups. In LC50, vacuoles in cytoplasm and
LC50 group showed swelling of the follicle walls and
degeneration of oocytes were observed. Similar changes
deterioration of cellular material. F ollicle wall was ruptured
were observed in both LC0 and LC50 groups of V. bengalensis
in places. Cytoplasm of oocytes showed prominent vacuoles.
A few vitellogenic oocytes showed karyolysis and appeared after acute exposure to folithion and lebaycid (Muley, 1985)
to undergo degeneration. The vitellogenic oocytes showed and in the female gonad of Meretrix meretrix after acute
fragmentation of cytoplasm. The damage was more severe exposure to Cypermethrin (Prabhupatkar, 2004).
than LC0 group (Fig. 3). Any particular alteration of cell may
Most of the study is related to pesticide induced changes in
indicate the presence of disease or the toxic substance. There the cytoarchitecture of different organs were carried out on
is a clear correlation between pathological condition of cell or fishes and other invertebrates, while that of mollusk and
tissues and its affected functions (Brown et al, 1967). The bivalves were still scanty (Shinde et.al, 2019). It is widely
extent of damage induced by the toxicant to a particular organ recognized that, growth and reproduction in aquatic animals
can also be judged on a cellular level. Pathological and were affected due to contamination of toxicants in aquatic
76 International Journal of Biological Innovations 1 (2), (DEC. 2019)

media. The pesticide shows the hazardous effect on the 9. Kandasamy D. and Muthukumaravel K. (2010).
inhabitants of the aquatic species. There were Toxic effect of chromium on the histoarchitectural
histopathological changes in Lamellidens corrianus and alterations of ovary of freshwater fish, Oreochromis
Lamellidens marginalis exposed to Cythion Malathion from mossambicus. J. Exp. Zool. India. 13(2): 413-414.
Godavari River, Paithan. The severity of effect of pesticide of
10. Khan M. and Jha B. S. (2000). Gonadal histopathology
both the species was more at LC50 than LC0 from gonads
of the freshwater fish, Channa punctatus under
(Muley and Mane, 1987).While studying alterations in gonad
phosalone exposure. Indian J. Env. and Ecoplan. 3: 587 –
due to Endosulfan to freshwater bivalve, Lamellidens
590.
marginalis and Lamellidens corrianus from Godavari River,
Paithan, similar trend of histopathological changes was 11. Kumari Anitha S. and Kumar Sree Ram N. (1997).
observed and it was stated that, the pesticide stress affect the Histopathological alterations induced by aquatic
reproductive status (Muley and Mane, 1998). While studying pollutants in Channa punctatus from hussainsagar lake
the effect of carbaryl on fish, Clarrius batrachus gonad (A.P.). J. Environ. Biol. 18(1): 11-16.
vacuolization and necrosis was observed in tissue (Jyothi and 12. Muley D. V. (1985). Effect of pollutants on freshwater
Narayan, 1999). Author observed similar changes in the molluscs from Godaveri river at Paithan. Ph. D. Thesis,
present investigations also.
Dr. B.A. Marathawada University, Aurangabad, M.S.
ACKNOWLEDGEMENTS India.1-25.
The author expresses her sincere thanks to the Dr. Maqdoom 13. Muley D. V. and Mane U. H. (1987). Sublethal effect of
Farooqui , Principal, Dr. Rafiq Zakaria College for Women, mercuric chloride on the tissue composite on of bivalve
Aurangabad for valuable input and good cooperation. mollusc, Lamellidens marginalis. Biol. Bull. of India.
1987. 9(1):31-40.
REFERENCES
14. Muley D. V. and Mane U. H. (1990). Histopathological
1. Baruah B. K. and Das M. (2002). Histopathological changes in body parts of the freshwater gastropod,
changes in ovary of fish, Heteropneustes fossilis exposed Viviparus bengalensis Lam. due to pesticides. J.
to paper mill effluent. Aquacult. 3(1): 29-32. Environ. Biol. 11: 413-425.
2. Bellas J., Beiras R. and Vazquez E. (2004). Sublethal 15. Muley D. V. and Mane U. H. (1998). Seasonal
effects of trace metals (Cd, Cr, Cu, Hg) on variations in the toxicity of Falithion and Lebaycid
embryogenesis and larval settlement of the ascidian freshwater gastropod Viviparous bengalensis (Lamarck)
Ciona intestinalis. Arch. Environ. Contam. Toxicol. 46
from Godavari River, Maharashtra state, India. Ad Bios.
(1):61–66.
7 (1):37-46
3. Brown V. M., Jordan D. H. M. and Tiller B.A. (1967).
16. Otitoloju A. A., Ajikobi D. O. and Egonmwan R. I.
The effect of temperature on the acute toxicity of phenol
(2009). Histopathology and bioaccumulation of heavy
to Rainbow trout in hard water. Water Research. 1 (8-9):
metals (Cu and Pb) in the giant land snail, Archachtina
587-594.
marginatai (Swainson). The open Environmental
4. Dholakia A.D. (2013). Identification of Marine and Pollution and Toxicology Journal. 1: 79-88.
Freshwater Molluscs Shells:1-186. (Published by Daya
Publishing house, A Division of Astral International Pvt. 17. Phirke P. P. (2008). Effect of thiodan and quinalphos on
Ltd., New Delhi-110 002). some physiological parameters of the freshwater
bivalve, Parreysia corrugata. Thesis submitted to Dr.
5. Hazarika R. and Das M. (1998). Toxicological impact Babasaheb Ambedkar Marathwada University,
of BHC on the ovary of the air breathing catfish, Aurangabad.
Heteropneustes fossilis. Bull. Environ. Contamin.
Toxicol. 60 (1):16-21. 18. Prabhupatkar M. M. (2004). Cypermetrin induced
toxicity to the estuarine clam, Meretrix meretrix
6. Jonnalagadda P. R. and Rao M. B. P. (1996). (Linnaeus). M.Sc. Thesis. University of Mumbai,
Histopathological changes induced by specific pesticide Mumbai. M.S. 1-60
on some tissues of the fresh water snail, Bellamya
dissimilis Müller. Bull. Environ. Contam. Toxicol. 57(4): 19. Ramchndra Mohan M. (2000). Malathion induced
648-654. changes in the ovary of freshwater fish, Glassobius
quaris (Ham.). Poll. Res. 19 (1): 73-75.
7. Jyothi B. and Narayan G. (1996). Effect of
organophosphorous insecticide phorate on gonads of 20. Rashatwar S. S. and Ilyas I. C. (1994). Effect of
freshwater fish, Clarius batrachus (Linn). Poll. Res. phosphomidon in a freshwater teleost fish Nemachelius
15(3): 293-296. denisonii Day - Histopathological and Biochemical
studies. J Environ Biol. 5: 1-8.
8. Jyothi B. and Narayan G. (1999). Certain pesticide
induced carbohydrate metabolic disorders in the serum 21. Shinde Nitin (2017). Malacological analysis of
of freshwater fish Clarias batrachus (Linn). Food. Godavari river of Western Maharashtra, India, Lap
Chem. Toxicol. 37(4): 417 421. Lambard Academic Publishing. 1-68p.
Shaikh Yasmeen 77

22. Shinde N.G., Gaikwad D.M., Vedpathak A.N. and 25. Waykar B. and Lomte V.S. (2004). Carbonyl induced
Bhupalwar S.R. (2019). Gemetogenic Changes in changes in the ascorbic acid content in different tissues
Freshwater Bivalve: Lamelledins corrianus in Monsoon of fresh water bivalve Parreysia cylindrica. Asian J.
due to removal of Cerebral Ganglia and Injection of its Micro Biotech and Environ Sci. 6(3): 181- 189.
Extract. International Journal on Biological Sciences. 26. Wester P.W. and Canton J.H. (1991). The usefulness of
10 (1): 39-4 histopathology in aquatic toxicity study. Comp Biochem.
23. Waykar Bhalchandra B. (1998). Effect of pesticides on Physiol. C. 100(1-2): 115-117.
some physiological activities of fresh water bivalve, 27. Zhou X., Upatham E. S., Kruatrachue M. and
Parreysia cylindrica. Ph.D. Thesis, Dr. Babasaheb Sretarugsa P. (1993). Effects of niclosamide and
Ambedkar Marathwada University, Aurangabad (M.S.) Eucalyptus camaldulensis on Biomphalaria glabrata,
India. the snail intermediate host of Schistosoma mansoni. J.
24. Waykar B. and Lomte V.S. (2002). Studies on Sci. Soc. Thailand. 19: 117-128.
histopathological changes in the gill of freshwater
Bivalve, Parreysia cylindrical exposure to endosulfan. J.
Aqu. Biol. 17(2): 67-72.