Research Journal of Chemistry and Environment_____________________________ Vol. 19 (4) April (2015) Res. J. Chem. Environ.

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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

*thyagarajen@gmail.com

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

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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

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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.

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(Received 14th August 2014, accepted 15

th November

2014)