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Research Journal of Chemistry and Environment_____________________________ Vol. 19 (4) April (2015) Res. J. Chem. Environ.
42
Copper sulphate induced histopathological alterations in organs of Ariopsis seemanni
(Colombian Shark Cat Fish) Thyagarajan R.*, Narendrakumar G., MasilamaniSelvam M. and Rameshkumar V.
Department of Biotechnology, Faculty of Bio and Chemical Engineering, Sathyabama University,
Chennai, 600119, INDIA
Abstract The effect of copper sulphate on liver, gills and
muscles of fish Ariopsis seemanni was observed as
metabolic indicators. The fish was exposed to sub-
lethal concentrations of copper sulphate for 5, 10 and
15 days at 2 ppm, 4 ppm and 6 ppm. The
histopathological study on the gills, liver and muscles
was also done in order to determine the tissue
damage. These findings suggest significant increase of
transaminase activity while reduced amount of
carbohydrates and protein content might be the
consequence of tissue damage in Ariopsis seemanni.
Keywords: Copper sulphate, Ariopsis seemanni,
histopathology, tissue damage.
Introduction Copper is highly toxic in aquatic environments and affects
on invertebrates, fishes and amphibians with all three
groups equally sensitive to chronic toxicity4,12
. Copper will
accumulate in many different organs in fish and mollusks.
While mammals are not as sensitive to copper toxicity as
aquatic organisms, biomagnification plays critical role in
their toxicity. Toxicity in mammals includes a wide range
of animals and affects in the form of liver cirrhosis,
necrosis in kidneys and the brain, gastrointestinal distress,
lesions, low blood pressure and fetal mortality2, 6,13,14
.
Study of histopathology is of prime importance in the
diagnosis, etiology and prevention of disease. In fishes, it is
observed that the external organs are affected due to toxic
chemicals, causing loss of equilibrium, increase in
opercular movement, to and fro irregular vertical
movements, finally leading to death. This may be attributed
to the significant damage to the internal organs.
Histopathological study thus gives us useful data
concerning tissue change prior to external manifestation3.
Histopathogical changes appear as a medium term response
to sub-lethal stressors and histology provides a rapid
method to detect effect of irritants, especially chronic ones,
in various tissues and organs5. The exposure of fishes to
chemical contaminants is likely to induce a number of
lesions in different organs7.
Material and Methods Catfish, Ariopsis semmani were purchased from Kolathur
fish market in Chennai, Tamilnadu, India. Fishes were
initially treated with 0.01% KMnO4 solution for 15 min to
avert any dermal infection before putting into the aquarium
tanks. Fishes were distributed in 100 L capacity aquarium
tanks and were kept for a period of 15 days for adaptation
to the lab condition.
The experimental design consisted of a control and three
treatments with different concentrations (2, 4 and 6ppm) of
copper sulphate (CuSO4.5H2O), two replicates per group
with twenty fishes in each replicate. Fishes showing no
respiratory movement and response to physical stimuli
were considered as dead and removed immediately11
.
Microscope Examination: After 5, 10 and 15 days of the
treatment, five fishes were taken from each tank. The gill
arches of the fish were excised from both sides. Fish were
dissected, the abdominal cavity was operated and the liver
and kidney were excised quickly and were fixed in Bouin’s
solution as histological fixative for 24 hours8. According to
Humason9, the specimens were processed as usual in the
recognized method of dehydration, cleared in xylene and
finally embedded in paraffin wax before being sectioned at
5µm using a rotary microtome (Leica RM2235
Germany)10
. The specimens were stained with hematoxyl in
and eosin. Finally, the prepared sections were examined
and photographically enlarged using light microscope
(Hamilton compound photomicroscope).
Results and Discussion Histopathology: The histopathological changes observed
in present investigation after exposure to lethal
concentrations of copper sulphate in the liver, gills and
muscles of freshwater fish Ariopsis seemanni have been
depicted in the photos from figures 1.1 to 3.4. The organs
of the fish exposed to heavy metal compounds exhibited
marked histopathological alterations.
Liver, gills and muscles of Ariopsis seemanni exposed to
lethal concentrations of copper sulphate showed
vacuolation in the cytoplasm, degeneration of nuclei
vacuolation in stroma. The alterations in liver of fish
exposed to sublethal concentration of copper sulphate were
cloudy swellings of the cells with large vacuoles,
degeneration of nuclei, vacuolation in stroma, disorganized
(disarray) hepatic cords, loss of shape of hepatocytes, pycnotic nuclei, shifting of nuclei on one side of the cell
and prominent necrosis.
Research Journal of Chemistry and Environment_____________________________ Vol. 19 (4) April (2015) Res. J. Chem. Environ.
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Fig. 1.1: Gills of fish at control Fig. 1.2: Gills of fish at 2 ppm
Fig. 1.3: Gills of fish at 4 ppm Fig. 1.4: Gills of fish at 6 ppm
Fig. 2.1: Heart of fish at control Fig. 2.2: Heart of fish at 2 ppm
Fig. 2.3: Heart of fish at 4 ppm Fig. 2.4: Heart of fish at 6 ppm
Research Journal of Chemistry and Environment_____________________________ Vol. 19 (4) April (2015) Res. J. Chem. Environ.
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Fig. 3.1: Liver of fish at control Fig. 3.2: Liver of fish at 2 ppm
Fig. 3.3: Liver of fish at 4 ppm Fig. 3.4: Liver of fish at 6 ppm
Fig. 4.1: Kidney of fish at control Fig. 4.2: Kidney of fish at 2ppm
Fig. 4.3: Kidney of fish at 4ppm Fig. 4.4: Kidney of fish at 6ppm
The changes observed in the muscles and gills due to
exposure to sublethal concentrations of copper sulphate
include rupture of blood sinusoids. Severity of damage was
more in the fishes with higher concentrations of copper
sulphate. It was also found to be dependent upon time of
exposure. These results were similar to Horne and Duson4,
Hadi and Alwan10
and Sindermann7.
Conclusion Heavy metals like copper enter the aquatic ecosystem
through a wide spectrum of natural source such as volcanic
eruption, soil erosion and anthropogenic activities
including use of pesticides, seepage as well as percolation
of industrial wastes leading to biomagnification in the food
chain. Histopathological alterations on Ariopsis seemanni
Research Journal of Chemistry and Environment_____________________________ Vol. 19 (4) April (2015) Res. J. Chem. Environ.
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under the influence of heavy metals can be used as a
sensitive model to monitor the aquatic pollution.
The histopathological changes observed in present
investigation after exposure to sub-lethal concentrations of
copper sulphate in the liver, gills, muscles and kidney of
freshwater fish Ariopsis seemanni have been depicted in the
photos from 1.1 to 4.4. The organs of the fish exposed to
copper sulphate exhibited enormous histopathological
alterations.
Gills, being the most exposed organ of the fish, liver being
the centre of metabolism and kidney being the filtration
unit have the effects of copper sulphate on it to a greater
extent. The damage done to the organs induced
histopathological alterations.
Reference 1. Adil A. Wani, Sikdar-Bar M. and Khan Hilal Ahmad, Acute
toxicity of copper sulphate to African catfish, (Clariasgariepinus),
GERF Bulletin of Biosciences, 4(1), 14-18 (2013)
2. ATSDR, Toxicological Profile for Copper, U.S. Public Health
Service, Agency for Toxic Substances and Disease Registry,
Atlanta, G.A. (1990)
3. Deore S.V. and Wagh S.B, Heavy metal induced
histopathological alterations in liver of Channagachua, Journal of
Experimental Sciences, 3(3), 35 (2012)
4. Horne M. T. and Dunson W. A., Effects of Low pH, Metals
and Water Hardness on Larval Amphibians, Archives of
Environmental Contamination and Toxicology, 29, 500-505
(1995)
5. Johnson L.L., Stehr C.M., Oslon O.P., Myers M.S., Pierce
S.M., Wigren C.A., McCain B.B. and Varanasi U., Chemical
contaminants and hepatic lesions in the winter flounder
(Pleuronectesamericanus) from the Northeast Coast of the United
States, Environmental Science and Technology, 27, 2759-2771
(1993)
6. Kabata-Pendias A. and Pendias H., Trace Elements in Soils and
Plants, 2nd ed., CRC Press, Boca Raton, 365 (1992)
7. Sindermann C.J., Pollution-associated disease and
abnormalities of fish and shellfish” a review, Fishery Bulletin, 76,
717-749 (1979)
8. Tao J. S., Liu C., Dawson Cao R. R. and Li B., Uptake of
particulate lead via the gills of fish (Carassiusauratus), Arch.
Environ. Contam. Toxicol., 37, 352-357 (1999)
9. Humason G.L., Animal tissue technique, Freemand W.H. and
Co., Sanfrancisco (1967)
10. Hadi A.A. and Alwan, Histopathological changes in gills,
liver and kidney of fresh water fish, Tilapia zillii, exposed to
aluminum, Int. J. of Pharm. & Life Sci., 3(11), 2071-2081 (2012)
11. Thyagarajan R., Keshav Vivek, Narendrakumar G.,
Masilamani Selvam M. and Rameshkumar V., Effect of Copper
Sulphate on Aspartate Aminotransferase (AST) and Alanine
Aminotransferase (ALT) Activity in Freshwater Fish Ariopsis
seemanni, Research Journal of Pharmaceutical, Biological and
Chemical Sciences, 6(1), 1219- 1224 (2015)
12. US EPA, Wildlife Exposure Factor Handbook, Vol. 1
EPA/600/R-93/187a (1993)
13. Vymazal J., Algae and Element Cycling in Wetlands, Lewis
Pub., Boca Raton, 689 (1995)
14. Patel R.M., Patel D.M., Shah K.P. and Patel D.A., Synthesis
of Polyketones and their Antimicrobial Study, Res. J. Chem.
Environ., 3(2), 47 (1999)
15. Ware G., Pesticides, Theory and Application, Freeman W.H.,
New York, 308 (1983).
(Received 14th August 2014, accepted 15
th November
2014)