Some biochemical variations during ovarian vitellogenic growth in hill-stream teleost Garra mullya due to sublethal concentration of cadmium has been discussed. Total protein, cholesterol and glycogen in ovary and liver along with gonadosomatic index (GSI) and hepatosomatic index (HSI) in Cd-treated fish exhibited significant decrease while liver glycogen remained unaltered

Khan, E.A.

Mehrotra, P.N.

Panday, P.N.

Saxena, N.

Sinha, M.P.

Aquatic Commons

Journal of the Indian Fisheries Association 
21, 1991, 11-U 
BIOCHEMICAL VARIATION DURING OVARIAN VITELLOGENIC GROWTH IN A 
HILL STREAM TELEOST GARRA MULLYA (SYKES)DUE TO CADMIUM TOXICITY 
EIISAN A. KHAN, M.P. SINHA, N. SAXENA, P. N. PANDA Y AND P. N. MEHROTRA 
P.G.Department of Zoology, Rm1chi University, Ranchi-834 008, Bihar. 
ABSTRACT 
Some biochemical variations during ovarian vitellogenic growth in hill-stream teleost Garra mul/ya due to sublethal 
concentration of cadmium has heen di'icussed. Total protein, cholesterol and glycogen in ovary and liver along with 
gonadosomatic index (GSI) and hepalosomatic index (IISI) in Cd-treated fish exhibited significant decrease while liver 
glycogen remained unaltered. 
INTRODUCTION 
Most of the fish species are seasonal breeders and 
the breeding period is marked by complex 
physiological processes in the body which result 
in remarkable biochemical changes in fish tissues 
specially liver and gonad. Ovarian vitellogenic 
growth is ini tiatcd by gonadotropins secreted 
by pituitary which stimulate the ovary to secrete 
estrogen. Much of the yolk material of eggs in fishes 
is synthesized by liver under the influence of 
estrogen, as a complex lipophospho-protein 
precursor known as vitellogenin (Emmersen and 
Emmersen, 1976; Tala, 1978). This is carried to 
the ovary by the blood to be incorporated there 
during vitellogenesis as a phosphoprotein-lipoprotein 
complex (Emmersen and Emmersen, 1976; Medda 
and Dus Mahaputra, 1980; Quinitio eta!., 1989) 
which eventually forms much of the dry matter of the 
ovary. As a result the phase of vitellogenic growth 
of fish is accompanied by changes in cellular 
constituents of liver and ovary, speciully protein, 
glycogen and cholesterol (Emmersen and Emmersen, 
1976; I3uno, 1977; Mcdda and Das Mahapatra, 1980). 
During the last few decades the multidimensional 
human activities has resulted in enormous 
poll uti on problem of water bodies from industrial, 
mining and other sources. Consequently heavy 
metal toxicity has b'ecomc an alarming problem 
for water bodies and cadmium is one of them. 
Cudmium has been found to affect the 
physiological and biochemical constitution of 
various fish species (Dull, 1967; Banerjee et at., 
1978; Wani and Latcy, 1982, 1983). However, the 
information about the effect of cadmium on 
biochemical composition of hill-stream fishes 
from Chhotanagpur pluteau is still inadequate 
though the rivers und streams of this region often 
get contaminated by heavy metals due to vmious 
industrial and mining efnuents. 
11 
In the present communication, an attempt has 
been made to study the effect of Cd-toxicity 
on the ovarian vitellogenic growth in a hill-stream 
teleost Carra Mullya which is a common 
food fish of this region specially in rural areas. 
MATERIAL AND METHODS 
Mature Carra mullya, (length, 14-15 em; average 
weight, 33.2 g) were used as test fish. Fishes 
were collected from survarnarekha river in the 
Getalsud region, 30 kms west of Ranchi town with 
the help of local fishermen and were acclimatized 
to laboratory condition for a week before start 
of the experiment. Probit analysis met-hod (Finney, 
1971; Fisher and Yates, 1974) was applied to 
detern~ine the 96 h Lc50 value. Experiments were 
carrieu' out during April, May and June separately 
which represents the period of maximum ovarian 
vitellogenic growth from the site mentioned above 
(Khan and Mehrotra, 1991). One dozen individuals 
were kept each in Cd-containing and control 
waters. Experimental group was lodged in water 
containing 4 mg!litre cadmium chloride (CdC1) 
on the second day of each month from April to June 
and were sacrificed on the 30th day of the respective 
months i.e. after 28 days of Cd-exposure. Control 
groups were maintained in ide'ntical conditions 
without CdC1 2• During experinwnt the fishes were 
fed with llydrilla leaves. Size of the aquarium 
was 40 em x 40 em x 40 em. The physico-chemical 
characteristics of water did not vary significantly 
during the three months of experiment and were 
: temperature 24.7 - 26.2° C; pli 7.2 - 7.5; total 
hardness 61 - 72 ppm; total alklinity 107 p - 116 
ppm and dissolved oxygen 6.5 - 7.5 ppm 
At the .end of experiment (28 days) in each 
month the liver and ovary were quickly dissected 
out and gonadosomatic index (GSI i.e. ovary wt./ 
100 g body wt.) und hepatosomatic index (IISI Le. 
12 KHAN, SINHA, SAXENA, PANDAY AND MEHROTRA 
liver wt./100 g body wt.) were determined. Tissues 
were transferred to chilled fish saline and . were 
processed for the estimation ·of total protein, 
cholesterol and glycogen. Methods of Munro and 
Fleck (1966) modified by Abalain et al. (1980) 
were applied for the extraction and the method 
of Lowry et a!. (1951) was applied for the 
estimation of protein. The method of Kabara 
(1962) was used for extraction and estimation of 
cholesterol. The method of Kemp et a!. (1954) 
modified by Krishnaswami and Srinivasan (1961) 
was applied for extraction and estimation of 
glycogen. Statistical analysis of the data was done 
by student's 't' test (Bruning and Kintz, 1977). 
RESULTS 
Results of the experiments are summarised in 
Tables I and II for liver and ovary respectively. 
Protein and cholesterol contents of liver and ovary 
were found to decrease significantly in Cd-treated 
group in comparison to control during April, May 
and June along with corresponding fall in HSI as 
well as GSI. Decrease of GSI was more significant 
in April and May (p<0.001) than in June (p<0.02). 
Similarly I-lSI exhibited more significant decrease 
in May and June (p<0.001) than April (p<0.05). 
Bothe liver and ovary proteins recorded highly 
significant fall (p<0.001) in Cd-treated specimens 
during all the three months of experiment. 
Cholesterol content also exhibited highly 
significant decrease during all three months in 
both liver and ovary of Cd-exposed specimens 
(p<0.001). Glycogen content recorded significant 
fall (p<0.001) in ovary of Cd-treated fish whereas 
in liver the change of glycogen was statistically 
insignificant. 
DISCUSSION 
The key product in vitellogenesis ts a 
multicomponent lipo-phospho-protein called 
vitellogenin which is synthesized in the liver and 
transported by blood to the ovary (Emmersen and 
Emmersen, 1976; Quinitio et a!. 1989). Before 
incorporation in the ovary as yolk granules, the 
vitellogenin is cleaved into yolk protein-phosvitin 
and lipovitellin (Tata, 1978). The process of 
vitellogenesis is initiated in liver under the 
influence of estro,e:en (Emmersen and Emmersen, 
1976; Bano, 1977; Medda and [)as Mahapatra, 
1980). Deposition of yolk protein in the ovary is 
reflected in the increased gonadosomatic index and 
hepatosomatic index (Emmersen and Emmersen, 
1976; Quinitio et al., 1989). A similar result of 
GSI and HSI increase was observed during the 
present study among con.trol group of fishes 
TABLE I :Variations in the protem content, cholesterol and glycogen in the liFer and of lrepatosomatic 
index in Carra mullya due to cadmium toxicity. 
Month 
April 
May 
June 
Total Protein 
(Percent) 
Control Treated 
28.50 18.10 
±0.37 ±0.29 
p<O.OOl 
26.40 
±0.31 
20.00 
±0.45 
p<0.001 
22.60 17.30 
+0.26 ±0.42 
p<0.001 
NS - Non Significant. 
Cholesterol 
(mg/g tissue) 
control Treated 
10.80 5-.90 
±0.26 ±0.22 
p<O.OOl 
11.50 6.50 
+0.22 ±0.42 
p<0.001 
11.60 6.20 
±0.28 ±0.29 
p<O.OOl 
Glycogen 
( mg/g tissue) 
control Treated 
5.70 5.40 
+0.29 ±0.21 
NS 
5.30 5.20 
+0.26 ±0.26 
NS 
5.40 5.70 
+0.21 ± 0.21 
NS 
Hepatosomatic 
Index (HSI) 
control Treated 
2.60 1.00 
+0.34 ±0.20 
p<0.05 
2.80 1.40 
± 0.26 ±0.15 
p<O.OOl 
2.10 1.40 
+0.17 ±0.15 
p<O.OOl 
BIOCHEMICAL VARIATION IN CARRA MULL YA 13 
TABLE II : Variations in the protein content, cholesterol and glycogen in the OVGI)' and of gonadosomatic 
index in Garra mullya due to cadmium toxicity. 
Month 
April 
May 
June 
Total Protein 
(Percent) 
Control Treated 
23.60 15.60 
+ 0.64 ± 0.24 
p<0.001 
26.80 19.70 
± 0.42 ± 0.50 
p<0.001 
28.30 22.90 
+ 0.45 ± 0.53 
p<0.001 
Cholesterol 
(mg/g tissue) 
control Treated 
9.70 4.90 
+ 0.23 ± 0.26 
p<0.001 
13.30 5.60 
± 0.49 ± 0.18 
p<0.001 
14.30 9.70 
± 0.29 ± 0.23 
p<0.001 
(Table I and ll) while a sharp decrease was 
noticed in the Cd-treated specimens probably 
due to histological dumage done by toxic impact 
of cadmium. A significant fall in the gonadosomatic 
index along with disappearance of mature oocytes 
and liver damage has also been noticed in 
Cd-trcated Garra mullya (Wani and Latey·, 1982, 
1983) which confirms the present finding. This 
change of GSI and IISI can be attributed to 
Cd-toxicity as cadmium shows high affinity with 
sulphydryl and hydroxyl groups and ligands 
containing nitrogen which is considered to be the 
mechanism of cadmium toxicity (Nilsson, 1970). 
Consequently the binding with such groups in 
biochemical systems of living tissues makes the 
effect of Cd-toxicity lethal even at low concetrations. 
The cholesterol, protein and glycogen contents of 
liver and ovary have been found to increase during 
breeding season in various species of fish(Emmersen 
and Emmersen, 1976; I3ano, 1977; Das Mahapatra 
and Medda, 1980, 1982; Sherni, 1981). Cholesterol, 
a major source of steroids during gonad 
maturation in fishes rcnccts the physiological state 
of ovary by its profiles in it as steroidogenesis via 
cholesterol precusor involves complex. enzyme 
systeri1s (Armstrong, 1968). lienee, the level of 
cholesterol becomes an index of functional state 
of ovary in fishes. During the present study the 
cholesterol, glycogen and protein levels along 
with HSI and GSI were found to be high both in 
the ovary and liver of control groups as the 
spawning months came nearer June (Table-1 and 2). 
Glycogen 
(mg/g tissue) 
control Treated 
6.20 3.60 
± 0.20 ± 0.14 
p<0.001 
7.50 4.80 
± 0.50 ± 0.21 
p<0.001 
7.80 5.70 
+ 0.09 ± 0.27 
p<0.001 
Jlepatosomatic 
Index (HSI) 
control Treated 
6.70 2.50 
± 0.66 ± 0.80 
p<0.001 
10.70 5.80 
± 0.06 ± 0.24 
p<O.OCll 
11.50 8.20 
± 0.99 ± 0.72 
p<0.02 
However, in Cd-treated specimens all the 
parameters exhibited significant decrease in both 
ovary and liver except liver glycogen· which 
remained unaltered, suggesting the possibility that 
cadmium toxicity either blocks hormone action 
or the complex enzyme systems responsible for 
vitellogenesis and steroidogenesis. An elevated 
level of serum glucose and depletion of liver 
glycogen among Cd-treated Clarias batrachus and 
Tilapia mossambica has been reported (Banerjee 
et a!., 1978). The retention of glycogen level even 
in the Cd-treated specimens as has been observed 
during the present study may, however, be attributed 
to the stress condition under which liver has 
a tendency to retain glycogen (Love, 1970). 
ACKNOWLEDGEMENTS 
One of the authors (E. A. Khan) gratefully 
acknowledges the University Grants Commission,. 
New Delhi, for Providing financial assistance in 
the form of a Senior Research Fellowship. N. Saxena 
thanks CSIR, New Delhi, for providing a Re~earch 
Associateshi p. 
REFERENCES 
Abalain, J, H., .Jego, 1,, and Valotaire, Y. ~980. 
Effect of 17- ~Estradiol on the DNA, RNA, protein 
contents and on the DNA, RNA polymerase in the 
mullerian duct of the immature female newt 
P leu rode les wa ltli i ( Michah ). Gen. C omp. Endocr ·no!. 
40 : 402-408. 
14 K [IAN, Sl NHA. ')AXENA. PANDA Y AND MEHRO'J RA 
Armstrong, D. T. 1968. Gonadotropins, ovarian 
metabolism and steriod biosynthesis. Rec. Prog. 
Harms. Res. 24 : 255-319. 
Ball, I. R.1967. The toxicity of cadmium to rainbow 
trout, Salmo .gairdneri. Wat. Res. 1 : 805-886. 
Banerjee, S. K., Dastidar, S. G., Mukhopadhyay, 
P. K. and Dehadrai, P. V. 197g. Toxicity of 
cadmium. A comparative study in the airhreathing 
fish, Clarias batrachus (Linn.) and in the non-
airbreathing one, Tilapia mossamhica (Peters). Ind. 
J. exp. Bioi. 16 : 1274-1277. 
Bano, Y.1977. Seasonal variations in the biochemical 
compoSition of C /arias batr'achus L. Proc. Ind. A cad. 
Sci. 85B : 147-155. 
Bruning, J, L. and Kintz, B. L. 1977. In : 
Computational Handbook of Statistics. Scott. 
Foresman and Co. Palo, Alto, Calif. 
Das Mahapatra, A. K. and Medda, A.K. 1982. 
Effect of estradiol dipropionate and testosterone 
propionate on the glycogen, lipid and water contents 
of vitellogenic and non-vitellogenic singhi fish 
H. fossilis (Bloch.). Gen. camp. Endocrinol. 48 : 
476-484. 
Emmersen, B. K. and Emmersen, J. 1976. Protein, 
RNA and DNA metabolism in relation to ovarian 
vitellogenic growth in the flounder, Platichthys flesus 
(L.) Camp. Biochem. Physiol. 55B : 315-321. 
Finney, D. J, 1971. Probit Analysis. Cambridge 
University Press, London 333 p. 
Fisher, R. A. and Yates, C. B. E. 1974. Statistical 
Tables. Longman, 46 p. 
Kabara, J, J, 1962. In : Methods of Biochemical 
Analysis. (Ed. by D. Glick). Wiley Intersciences, New 
York. 10 : 263 p 
Kemp, A. and Anddenne, J, M. R. V. R. 1954. 
Biochem. J. 56 : 646 
Khan, E. A. and Mehmtra, I,, N. 1991. Seasonal 
Variations of protein, DNA and RNA in the liver 
and ovary of a hill stream and teleost Garra mullya 
(sykes). Proc. Wat. Symp. Freshwat Aquacult : 
84-85 pp 
Krishnaswami, S. and Srinivasan, V. V. 1961. 
Curr. Sci. 30 : 353. 
Love, R. M. 1970. The Chemical Biology of Fishes. 
Academic Press, London. 
Lowry, D. H., Rosenhrough, N.J., Farr, A. L. anJ 
RandalL R. J, 1951. Protein measurement with the 
folin phenol reagent. J. Bioi. Chern. 193 : 265-275. 
Medda, A. K. and Das Mahapatra, A. K. 1980. 
I :ffcct of estrogen and testosterone on the protein and 
nucleic add contents of liver, muscle and gonad and 
plasma protein content of male and female vi tellogenic 
and Jl<ln-vitellogenic singhi fish (H. fossilis). 
Gen.Comp. Endocrinol. 42 : 427-436. 
Munro, H. N. and Fleck, A. 1966. The 
determination of nucleic acids. In : Methods of 
Biochemical Analysis. (Ed.) D. Glick. Wiley 
Intersciences, New York. 14 : 113-176 
Nilsson, R. 1970. Aspects on the toxicology of 
cadmium and its compounds. Nat!. sci. Res. Cowt. 
Stockholm. 
Quinitio, G.F., Takemura, A. and Goto, A. 
1989. Ovarian development and changes in serum 
vitellogenin level in the river sculpin, coitus 
hangiongensis during annual reproductive cycle. 
Bull. Fac. Fish Hokkaido Uni. 40(4) : 246-253. 
Sherni, K. D. 1981. Seasonal changes in the 
biochemical composition of ovary in Heteropneustes 
fossilis (Bloch.). Fish Techno!. 18 : 101-103. 
Tata, J, R. 1978. Induction and regulation of 
vitellogenin systhesis by estrogen. In: Biochemical 
Action of Hormones (Ed. by G. Litwack). Academic 
Press, New York. 5, pp. 379-431, 
Wani, G. P. and Latey, A. N. 1982,. Cadmium 
toxicity of gonads in a teleost fish, Garra mullya 
(Sykes). Poll. Res. 1 : 39-44. 
Wani. G. P. and Latey, A. N. 1983. Toxic effects 
of cadmium on the liver of a freshwater teleost, 
Garro mullya (Sykes). Curr. Sci. 52(21): 
1034-1035. 


Biochemical variation during ovarian vitellogenic growth in a hill stream teleost Garra mullya (Sykes) due to cadmium toxicity

http://aquaticcommons.org/15933/1/JIFA21_011.pdf

Biochemical variation during ovarian vitellogenic growth in a hill stream teleost Garra mullya (Sykes) due to cadmium toxicity

Abstract

Similar works

Full text

Available Versions

Aquatic Commons