International

Volume I Number Journal

2 2010 for
[88 Environmental
- 93] Rehabilitation and Conservation
Volume I No. 2 2010
[ISSN 0975 - 6272] [94 - 98] [ISSN 0975
Bahuguna et al. - 6272] 

 
Effect of Heavy Metal Nickel on Enzymological Parameters of Cirrhina Mrigala

Arya, Shveta1, Matta, Gagan2, Sharma, H. B.3 and Singh, Jyotsna4

Received: April 20, 2010  Accepted: October 29, 2010  Online: November 30, 2010

Abstract Introduction
The aim of the present investigation is to study
Heavy metals are common persistent
the chronic toxicity of heavy metal Nickel on
pollutants of aquatic ecosystem entering them
some enzymes of carbohydrate metabolism in
through numerous diverse anthropogenic and
tissues of fresh water fish Cirrhina mrigala.
natural sources (Moore, 1991). Many
Fish were exposed to sublethal dose of Nickel
industrial and agricultural processes have
chloride (NiCl2) for 40 days. The first batch of
contributed to the contamination of fresh water
fish was sacrificed after 20 days of exposure to
systems thereby causing adverse effects on
sub lethal dose of nickel and the second after
aquatic biota and human health (Rose et al.,
40 days. The tissue extracts from liver, muscle,
1999). Heavy metal contamination may have
gills were taken and tested for the activity of
devastating effects on the ecological balance of
key enzymes of glycolysis and Kreb’s cycle.
the recipient environment and a diversity of
Increase in the activity of G-6-Pase in liver and
aquatic organisms (Farombi, et al., 2007;
gills shows increased rate of glycogenolysis to
Vosyliene and Jankaite, 2006; Ashraj, 2005).
meet energy demands of body. Increase in
Over a long period, the pollutants present in the
activity of LDH and inhibition in activity of
environment at very low levels may
PDH and SDH in liver, muscle and gills
accumulate within the body of aquatic species
indicate shift in metabolism from aerobic to
by various mechanisms to the extent that they
anaerobic.
exert toxic effects (Palaniappan and
Key words: Tissues | Enzymes | Nickel | Karthikeyan, 2009).
Cirrhina mrigala
Anthropogenic activities (i.e. mining,
For Correspondence: electroplating & steel plant operations) can
1 Singhania
2
University, Pacheri Bari, Rajasthan; India. result in nickel discharge into water and air
Department of Zoology and Environmental Science,
Gurukula Kangri University, Haridwar, India (Galvin, 1996). The nickel is an essential
3 Department of Zoology, B.S.A. (PG) College, Mathura (UP);

India element at low concentrations for many

4
Department of Zoology, K.L. Mehta Dayanand College for organisms; it is toxic at higher concentrations
Women, Faridabad, Haryana

Effect of Heavy Metal Nickel on Enzymological Parameters of Cirrhina Mrigala 

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Volume I Number 2 2010 [94 - 98]
ISSN: 0975 - 6272
(Clark and Keasling, 2002). Fish residing in
polluted freshwater systems are exposed to Ni,
 primarily, through the ingestion of
contaminated food and sediments (Dallinger
and Kautzky, 1985). The heavy metal in the
tissue of fishes may cause various
physiological defects and mortality (Torres et
al., 1987). Hence, the present investigation has
been carried out to study the effect of nickel on
enzyme activities in the various tissues of
freshwater fish Cirrhinus mrigala on chronic
exposure for 40 days.
Materials and Method
The fish, Cirrihina mrigala were purchased
from the local fish market having an average
length 12±3 cm and weight 200±2 gms The
fish were then kept in different aquaria for
conduction of various experiments. The fish
were acclimatized to laboratory conditions in
aquaria for a few days. In one aquarium the fish
were kept as control specimens given the same
food and environment as that of the
experimental fish except that they were not
given the dose of heavy metal compound.
Inorganic salt of heavy metal nickel namely
nickel chloride anhydrous (NiCl2) was the
experimental toxicant. To observe the chronic
effects of nickel, sublethal dose (1/10
concentration of 96 hr LC50) of the heavy metal
compound was given for 40 days. The first
batch of fish was sacrificed after 20 days of
exposure to sub lethal dose of nickel and the
second after 40 days. During exposure period
the fish were conditioned to feeding on packed
fish food at the rate of 2% of body weight. The
fish were fed once daily at 11am.
Effect of Heavy Metal Nickel on Enzymological Parameters of Cirrhina Mrigala
For the estimation of activities of enzymes of
glycolysis and kreb’s cycle, 10% of W/V
homogenates were prepared in 0.25 M sucrose
solution for tissues namely liver, gills and
muscles. The homogenates were centrifuged at
1000x g for 20 min. and clear supernatant
fluids were used as the source of enzymes. The
activities of Lactate Dehydrogenase(LDH),
Pyruvate Dehydrogenase(PDH), Succinate
Dehydrogenase(SDH) enzymes were
estimated by the triphenyltetrazolium chloride
method of Srikantan and Krishnamoorthi
(1995). The activity of Glucose-6-Phosphatase
was determined by adopting the method of
Swanson (1955). The significance of the
difference between control and experimental
means was calculated by Students‘t’ test
(Wardlaw, 1985).
Results and Discussion
Liver (Table-1)
A progressive increase was observed in the
activity of Glucose-6-Phosphatase ranging
from 38.03% to 62.65% during the entire
exposure period. The activity of lactate
dehydrogenase (LDH) increased after 40 days
by 25.46% but decreased after 20 days by
26.16%. Reduction was observed in the
activity of pyruvate dehydrogenase (PDH)
after 40 days however insignificant change
was observed after 20 days of exposure. The
activity of succinate dehydrogenase (SDH)
increased after 20 days of exposure whereas
decrease of 14.68% was observed in the
enzyme activity after 40 days of exposure.
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ISSN: 0975 - 6272
Gill (Table-2)
Progressive increase was noticed in the activity
 of Glucose-6-Phosphatase after 20 and 40 days
of exposure by 10.17% and 37.73%
respectively. The activity of lactate
dehydrogenase (LDH) increased by 35.93%
after 20 days and 54.00% after 40 days of
exposure. Reduction was observed in the
activity of Pyruvate dehydrogenase (PDH) and
Succinate dehydrogenase (SDH) upto 40 days
of exposure period.
Muscle (Table-3)
The activity of lactate dehydrogenase(LDH)
increased initially by 17.68% after 20 days and
was insignificant after 40 days of exposure.
The activity of pyruvate dehydrogenase (PDH)
decreased upto 40 days of exposure. The
succinate dehydrogenase (SDH) activity
decreased after 20 days (35.05%), was
insignificant after 40 days.
On exposure to nickel, different metabolic
alterations produced in different tissues varied
with period of exposure. Increase in the
activity of Glucose-6-Phosphatase in liver and
gills in all stages of exposure shows active
breakdown of glycogen reserve or increased
rate of glycogenolysis to meet energy demands
ENZYMES
GLUCOSE-6-PHOSPHATASE a
LACTATE DEHYDROGENASE b
PYRUVATE DEHYDROGENASE b
SUCCINATE DEHYDROGENASE b
Values are mean ±SD; n=6, NS= not significant
*significant, *p<0.05, **p<0.01, ***p<0.001
a
µg inorganic phosphate/mg Protein/hr
b
µg formazon/mg Protein/hr
Effect of Heavy Metal Nickel on Enzymological Parameters of Cirrhina Mrigala
of body. Dange (1986a) reported increase in
glycogenolysis in Sarotherodon mossambicus
exposed for 96 hr to a variety of chemicals
such as mercury, copper, naphthalene and
phenol.
Increase in the activity of SDH and decrease
in activity of LDH in liver after 20days shows
that in fish, the rate of oxidative metabolism
is increased to withstand the toxic stress.
However on chronic exposure for 40 days
there was significant increase in activity of
LDH and inhibition in activity of PDH and
SDH in liver indicating shift in metabolism
from aerobic to anaerobic. Inhibition in the
activity of LDH noted by Rajeshwari et al
(1990) in liver of Channa punctatus acutely
and chronically exposed to cadmium lends
support to the present findings.
Anaerobic metabolism was favoured in gills
(20, 40 days) and muscle (20 days) and in
liver after 40 days to meet energy demands.
Evidence in support to this comes from
increase noted in the activity of LDH and
decrease in the activity of PDH and SDH.
Sastry and Rao (1981) observed decrease in
LDH activity in liver, gills and Kidney of
Channa punctatus exposed to mercury.
CONTROL 20 DAYS 40 DAYS
233.39±0.02 322.16±0.01*** 379.63±0.01***
4.79±0.04 3.16±0.03*** 5.37±0.05***
3.14±0.05 3.18±0.06NS 2.45±0.02***
5.09±0.01 5.89±0.02*** 4.30±0.07***
Table 1: Alterations in Liver Enzyme Activities in
Cirrhina Mrigala Exposed to Nickel (Ni)
For 40 Days
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ENZYMES CONTROL 20 DAYS 40 DAYS

GLUCOSE-6-PHOSPHATASE a 27.72±0.05 30.54±0.04*** 38.18±0.03***
LACTATE DEHYDROGENASE b 4.87±0.03 6.62±0.06*** 7.50±0.07***
 
PYRUVATE DEHYDROGENASE b 3.73±0.01 2.06±0.03*** 1.79±0.04***
SUCCINATE DEHYDROGENASE b 6.49±0.04 5.05±0.02*** 4.79±0.04***
Values are mean±SD; n=6, NS= not significant
*significant, *p<0.05, **p<0.01, ***p<0.001 Table 2: Alterations in Gill Enzyme Activities in Cirrhina
a
µg inorganic phosphate/mg Protein/hr Mrigala Exposed io Nickel (Ni) For 40 Days
b
µg formazon/mg Protein/hr

ENZYMES CONTROL 20 DAYS 40 DAYS

LACTATE DEHYDROGENASE b 8.03±0.02 9.25±0.04*** 7.76±0.19NS

PYRUVATE DEHYDROGENASE b 11.05±0.04 4.01±0.06*** 5.30±0.06***

SUCCINATE DEHYDROGENASE b 4.51±0.03 3.15±0.02*** 4.40±0.11NS

Values are mean±SD; n=6, NS= not significant
*significant, *p<0.05, **p<0.01, ***p<0.001
b
µg formazon/mg Protein/hr
Table 3: Alterations in Muscle Enzyme Activities in
Cirrhina Mrigala Exposed to Nickel (Ni) For
40 Days

Conclusion Dallinger, R. and Kautzky, H. (1985): The

From the results obtained in the present study
it can be concluded that chronic exposure of
heavy metal nickel in fresh water fish Cirrhina
mrigala led to shift in the metabolism from
aerobiasis to anaerobiasis to combat the toxic
stress.
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