METALLOTHIONEIN ASSAY FOR BIO-MONITORING TRACE METAL

STRESS IN NILE CICHLID (OREOCHROMIS NILOTICUS) THRIVING IN

CONTAMINATED WASTEWATER-FED FISHPONDS IN EAST CALCUTTA
WETLANDS, A RAMSAR SITE

U. S. Roy1*, B. Chattopadhyay1 and S. K. Mukhopadhyay2

1
Ecotoxicology Laboratory, Government College of Engineering and Leather Technology, LB-III, Salt Lake City,
Kolkata- 700098, West Bengal, India.
2
Hoogly Mohsin College, Chinsurah 712101, West Bengal, India.

Abstract

Metals are serious chemical toxicants that disturb the environmental homogeneity by prolonged

environmental persistence and bioaccumulation. Bioaccumulation of both bioactive and non-bioactive

metals above thresh hold level often results in stress which leads to irreversible physiological conditions.

The present investigation was carried out to measure the amount of potential stress that fish species are

facing that thrive in wastewater-fed fishponds (WFFs) in East Calcutta Wetlands (ECWs), using

metallothionein as a biomarker and Oreochromis niloticus as suitable aquatic model. Levels of trace metals

(Cr, Mn, Fe, Cu, Zn and Pb) and the respective metallothionein levels along with total protein content were

measured in the liver, gill, muscle and brain of mature, healthy fish specimens, collected from both WFFs

and distantly located freshwater-fed fishponds (FFFs). Water samples from both qualities of fishponds were

analyzed for physico-chemical parameters including estimation of trace metals. The trend for trace metal

distribution in WFFs was Zn > Fe > Mn > Cr > Cu > Pb; and for FFFs it was Fe > Zn > Mn > Cu > Cr >

Pb. The levels of trace metals ranged between 4.690.67 mg L -1 (Zn) 0.320.10 mg L -1 (Pb) in WFFs and

4.580.72 mg L-1 (Fe) 0.120.02 mg L -1 (Pb) in FFFs respectively. The order of concentration of the

metals in fish organs collected from WFFs was as follows: Chromium Liver > Gill > Muscle > Brain;

Manganese- Liver > Gill > Brain> Muscle; Iron- Liver > Gill > Brain> Muscle; Copper- Liver > Gill >

Muscle > Brain; Zinc- Gill > Liver > Muscle > Brain; Lead- Liver > Gill > Muscle > Brain. The order of

concentration of the metals in fish organs collected from FFFs was as follows: Chromium Liver > Gill >

Muscle > Brain; Manganese- Liver > Gill > Brain> Muscle; Iron- Liver > Gill > Brain> Muscle; Copper-

Liver > Muscle > Gill> Brain; Zinc- Gill > Liver > Muscle > Brain; Lead- Brain > Liver > Gill > Muscle.

The highest concentration of trace metal were recorded in livers of fishes collected from both sites

4537.79507.12 mg Kg-1 dw (Fe) and 2748.33228.17 mg Kg-1 dw (Fe) for WFFs and FFFs respectively
whereas the lowest concentration were recorded in brain 0.860.10 mg Kg -1 dw (Zn) for WFFs and in

Muscle 0.170.02 mg Kg-1 dw (Pb) for FFFs respectively. ELISA assay revealed highest metallothionein

concentration in livers 344.7471.94 g g-1 ww and 54.5214.48 g g-1 ww whereas lowest concentrations

were recorded in brains 27.7043.28 g g-1 ww and 1.460.35 g g-1 ww for fishes collected from WFFs

and FFFs respectively. These findings are with well accordance to the previous works where liver, almost

invariably, always found to accumulate more metals and as ambient metal concentrations has been reported

to result in higher expression of metallothionein, it might as well resulted in the present investigation to

record a significantly higher metallothionein concentration in liver. Metallothionein thus acts in a manner to

detoxify the effects of toxic metals and probably maintains the proper metal homeostasis to lessen the

stress, otherwise intolerable.

*Corresponding author: srutpal@gmail.com; srutpal@yahoo.com