Two synthetic androgenic steroids, Ethylestrenol (17 β - Hydroxy - 17 α ethyl - estr - 4 - en - 3 - one) and Stanozolal (17 β - Hydroxy- 17 α - methyl - 5 a - androstano - 3, 2 - C - pyrazole) were fed via diet at 3 ppm to the spawn of Rohu and Mrigal which were reared up to fry stage over a period of 15 days in earthen carp nurseries. Both hormones enhanced growth of spawn. A maximum of 25.78% increase in length and 25.69% increase in weight as compared to the controls has been recorded. Growth rate was recorded to be 0.8 mm & 2.48 mg/day (control), and 1.13 mm & 2.67 mg/day (Stanozolol treated) in case of Mrigal spawn; and 0.91 mm & 2.39 mg/day (control), 1.12 mm & 2.90 mg/day (Ethylestrenol treated), and 1.10 mm & 2.57 mg/day (Stanozolol treated) in case of Rohu spawn. A decrease in the values of Relative Condition Factor upon hormone administration was also noticed

Bora, U.

Rao, P.S.

Venkateshvaran, K.

Aquatic Commons

Journal of the Indian Fisheries Association 
27,2000,27-35 
27 
FIELD APPLICATION OF ANABOLIC STEROIDS IN CARP SEED 
PRODUCTION 
II. REARING OF SPAWN TO FRY STAGE* 
BORA, U.,*** RAO, P.S., & VENKATESHVARAN, K.** 
Central Fisheries Extension Training Centre (CIFE) 
Kahinada - 533 007 A.P. 
ABSTRACT 
Two synthetic androgenic steroids, Ethylestrenol (17 ~ - Hydroxy - 17 a 
ethyl- estr - 4 - en- 3 - one) and Stanozolal (17 ~ -Hydroxy- 17 a -methyl - 5 a -
androstano - (3, 2 - C) - pyrazole) were fed via diet at 3 ppm to the spawn of 
Rohu and Mrigal which were reared upto fry stage over a period of 15 days in 
earth ern carp nurseries. Both hormones enhanced growth of spawn. A maximum 
of 25.78% increase in length and 25.69% increase in weight as compared to the 
controls has been recorded. Growth rate was recorded to be 0.8 mm & 2.48 n1g/ 
day (control), and 1.13Inm & 2.67 mg/day (Stanozolol treated) in case ofMrigal 
spawn; and 0.91 n1m & 2.39 mg/day (control), 1.12 mm & 2.90 mg/day (Ethylestrenol 
treated), and 1.10 mm & 2.57 mg/day (Stanozolol treated) in case of Rohu 
spawn. A decrease in the values of Relative Condition Factor upon hormone 
administration was also noticed. 
Keywords : anabolic steroids, carp nursery 
INTRODUCTION 
Many workers have used steroid 
hormones and their derivatives to increase 
growth rate and feed conversion efficiency 
in various species offish (Donaldson et al. 
1979, Higgs et al., 1982) and some amount 
of work has also been done in India (Deb 
and Varghese, 1988; Mogale, 1991; 
Omprakash; 1991; Venkateshvaranet al., 
1992). Despite these works, much remains 
to be explored on aspects of hormonal 
enhancement of the growth of the various 
life history stages of the Indian Major and 
Exotic carps. Moreover, most of these studies 
are in vitro and information on the 
effectiveness of these compounds under 
actual field conditions are wanting. The 
present in situ study was therefore taken 
up to assess the anabolic activities of two 
synthetic androgenic hormones on carp 
spawn during the nursery phase. 
MATERIAL AND METHODS 
i) Study Area : 
The study was conducted during the 
induced breeding season (July-August, 
1991) at the Freshwater Fish Farm of CIFE 
at Balabhadrapuram, Andhra Pradesh. 
Based on a dissertation submitted by the Senior author. 
** 
*** 
Present Address: Central Institution of Fisheries Education, Versova, Mumbai-400 061. 
Present Address: Dept. Fisheries (Assam), Mandhpur, Nagaon-782 001. 
28 BORA, RAO AND VENKATESHV ARAN 
ii) Test Hormones : not be maintained due to constraints of 
Based upon their ready availability in 
the market in tablet form at inexpensive 
rates, and on their established anabolic 
properties, the two following hormones were 
selected and used : 
a) Ethylestrenol : 17 ~ - Hydroxy- 17 
a- ethyl- estr- 4- en- 3- one. (Orabolin® 
ofM/s Infar (India) Limited, Calcutta). 
b) Stanozolal : 17 ~ - Hydroxy- 17 a -
methyl - 5 a - androstano - (3,2 - C) -
pyrazole. (Menabol® of M/s CFL 
Pharamaceuticals Limited, Bombay). 
iii) Test Animals : The spawn ofRohu, 
Labeo rohita and of Mrigal, Cirrhina 
mrigala were the test organisms which 
were reared upto fry stage in earthern 
nurseries over a period of 15 days. 
iv) Experimental Design: 
A total of 6 earthen nursery ponds, 
each measuring 20m x 10m (0.02 ha) and 
designated KNP 16, 20, 22, 23, and 25 
were involved in the present study. 
Details of stocking and artificial feeding 
are provided in Table 1. Replicates could 
training programme and commercial seed 
production activities of the farm. 
Pre and Post-stocking 
management procedures including artificial 
feeding were based on the CIFE Method 
(Venugopal and Venugopal, 1988). The 
hormonal tablets were finely powdered and 
premixed to the feed at 3 mg/kg offeed to 
achieve a uniform dosage of 3 ppm; this 
dosage was decided upon based on earlier 
works (Matty, 1985; Venkateshvaran et 
aL) 1993). 
v) Water Quality: 
Water quality parameters were 
monitored every morning and plankton 
population estimated every 5 days following 
standard methods (APHA, 1975),. 
vi) Growth studies and Relative 
Condition Factor: 
Growth of spawn was measured on the 
5th, lOth and 15th days from stocking and 
the fry harvested on the 15th day by 
repeated drag netting with 1/16" cotton 
dragnet. 
Table 1 : Showing details of stocking and hormone treatment via artificial feed 
in the 6 nursery ponds under study. 
Pond Species No. Stocked Av. Size stocked Date Hormone Date 
No. inlakhs Stocked Received Harvested 
16 Mrigal 1.2 7.0 mm- 1.5 mg 26.7.91 Ethylestrenol 9.8.91 
20 Mrigal 1.2 7.0 mm- 1.5 mg. 26.7.91 Stanonolal 9.8.91 
24 Mrigal 1.2 7.0 mm- 1.5 mg. 1.8.91 Nil 16.8.91 
22 Rohu 2.0 5.0 mm- 1.5 mg. 19.7.91 Ethylestrenol 2.8.91 
23 Rohu 2.0 5.0 mm- 1.5 mg 21.7.91 Stanozolal 4.8.91 
25 Rohu 2.0 5.0 mm- 1.5 mg 22.7.91 Nil 5.8.91 
FIELD APPLICATION OF ANABOLIC STEROIDS IN CARP SEED PRODUCTION 29 
The relative condition factor KTL CKTotal 
Length) was computed separately for each 5 
days set of length-weight data following 
Lagler (1956) and using the formula 
w K = --------
TL n 
a.L 
RESULTS 
The observed range of variations in 
physico-chemical parameters of water is 
presented in Table 2. In all the ponds 
plankton sediment volumes, from 0.2 to 
0.5 cc/50 a on day 0, rose to around 1.0 cc 
by the 5th day and remained between 1.0 
and 1.5 cc during the subsquent period. 
Anacystis sp. dominated the phytoplankters 
while Brachinus sp. and Keratella sp. 
dominated the zooplankton population. 
Results of the growth studies are 
presented in Table 3 as also Figs. 1 and 2. 
Table 4 presents the results of relative 
condition factor estimations. 
DISCUSSION 
i) Physico-chemical and biological 
P.arameters ofwater: 
The fluctuations observed in the 
physico-chemical parameters of water are 
characteristic of eutrophicated tropical 
ponds associated with aquacultural 
activities (Stickney, 1979). The presence of 
large quantities of cyanophyceans and 
rotifers in the plankton biomass point 
towards the productive nature of the water 
bodies studied (George et al.) 1986). 
ii) Ethyles·crenol : 
Ethylestrenol has enhanced the growth 
ofthe spawn of both Rohu and Mrigal, with 
a higher daily growth increment (Table 3; 
Figs. 1 and 2). Growth as recorded on the 
5th day was more in the control than in the 
ponds receiving treated feeds' but the trend 
reversed subsequently. Ethylestrenol has 
been shown to enhance growth when fed 
via the diet in Tilapia melanopleura 
(Hutchinson and Campbell, 1964) in the 
Table 2 :Showing range of variations in physico-chemical parameters of water 
during the study period. Data pooled up for 6 ponds over a period 
of 15 days. 
Parameter Mean with S.D. 
Temperature 
Air 29.5 ± 2.0°C 
Water 27.1 ± 2.2°C 
Dissolved Oxygen Content 6.8 ± 2.4 ppm 
Free Carbon Dioxide Content 4.8 ± 3.0 ppm 
pH 8.4 ± 0.4 
Total Alikalinity 134.3 ± 14.3 ppm CaC03 
30 BORA, RAO AND VENKATESHVARAN 
Atlantic salmon and Rainbow trout at 2.5 
ppm (Simpson, 1976a) and in the fry of 
Rohu, and Silver Carp at 3 ppm 
(Venkateshvaran et al. 1992) but it retarded 
growth of Catla fry at 3 ppm 
(Venkateshvaran et al. 1992). Matty (1985) 
also reported that ethylestrenol enhances 
gTowth in carps and salmonids. 
iii) Stanozolal : 
Stanozolal at 3 ppm in the present 
study enhanced growth of the spawn of 
both Rohu and Mrigal with better rate of 
Claily growth increment; growth increment 
improved over the control only after the 
5th day in this case also (Table 3, Fig. 1 
and 2) Stanozolal had been shown to 
enhance growth in carps and salmonids by 
Matty (1985), in the gold fish Carassius 
auratus by Bulkley and Swihart (1973) 
and in the fry ofMrigal and Common Carp 
by Omprakash (1990). 
Venkateshvaran et al., ( 1992) reported 
growth retardation in the fry of Catla, Rohu, 
and Silver carp at dietary stanozolallevels 
of 3 ppm whereas the spawn ofRohu in the 
present case has grown better at the same 
dose. Anabolic responses in tllis case and in 
the case of Catla with Ethylestrenol may 
be size-related as the works of Simpson 
(1976 b) would suggest and further detailed 
investigations are needed in this regard. 
Table 3 : Showing growth of spawn in length (mm) and weight (mg) over a 
period of 15 days at intervals of 5 days, growth rate per day, and 
percentage of growth as compared to controls. 
A:MRIGAL 
Pond Hormone ODay 5Day 10Day 15Day Increment %Growth 
per day over control 
KNP24 Nil 7.0mm 11.0mm 12.7 mm 19.0 mm 0.80mm 
(Control) 1.5 mg 10.0mg 14.0mg 38.6mg 2.48mg 
7.0mm 9.5mm 16.8 mm 22.8mm 1.05 mm 20.00 
1.5 mg 6.4mg 41.5 mg 46.3mg 2.99mg 19.95 
KNP 16 E 
7.0mm 9.0mm 16.7mm 23.9mm 1.13 mm 25.78 
1.5mg 6.1mg 28.0mg 41.5 mg 2.67mg 7.51 KNP20 S 
B:ROHU 
KNP25 Nil 5.0 nun 12.0mm 13.1 mm 18.7mm 0.91 mm 
(Control) 1.5mm 12.0mg 16.8 mg 35.8mg 2.39mg 
KNP22 E 5.0mm 8.0mm 15.0mm 21.8 mm 1.12 mm 16.58 1.5 mg 4.0mg 42.0mg 45.0mg 2.90mg 25.69 
KNP23 s 5.0 mm 9.0 mm 14.4mm 21.5 mm 1.10 mm 14.97 
1.5 mg 5.0mg 24.0mg 40.0 mg 2.57mg 11.73 
E = Ethylestrenol, S = Stanozolal 
FIELD APPLICATION OF ANABOLIC STEROIDS IN CARP SEED PRODUCTION 3 1 
25 
;?0 
E 
E 
c 
-- 15 
.c 
01 
c 
., 
....J 
10 
5 
5 
I 
I 
I 
I 
I 
I 
; I I .... ' 
1:,/ 
. I 
Days 
I 
I 
I 
I 
I 
;' 
/ 
/ 
Cirrhilw mrigala 
---
, 
;' 
/ 
-·E 
.. .--
15 
Fig. 1: Showing growth of Mrigal spawn over a period of 15 days in the control pond 
and in ponds receiving hornwne-treated feed. 
C: Control; E : Ethylestrenol; S : Stanozolal 
32 
E 
E 
.c 
-[ 
....J 
-.c 
Vi 
BORA, RAO AND VENKATESHV ARAN 
s 
Oayfi. 
Labeo rohita 
..-··-E 
-·· ,,--·· 
. .....s 
/ " 
,I 
10 
" 
" 
" 
" 
./ 
" 
" / 
./ 
/ t 
lS 
Fig. 2 :Showing growth of Rohu spawn over a period of 15 days in the control pond 
and in ponds receiving hormone-treated feed. 
C: Control; E : Ethylestrenol; S : Stanozolal 
FIELD APPLICATION OF ANABOLIC STEROIDS IN CARP SEED PRODUCTION 33 
Table 4: Showing relative condition factor <KrotalLength) of spawn over a 
period of 15 days computed on the 5th, lOth & 15th days. 
Pond Species Hormones 
KNP24 MRIGAL Nil 
KNP16 MRIGAL E 
KNP20 MRIGAL s 
KNP25 ROHU Nil 
KNP22 ROHU E 
KNP23 ROHU s 
E = Ethylestrenol, S = Stanozolal 
iv) Relative condition factor: 
A depression in the relative condition 
factor has been observed (Table 3) indicating 
increase in weight(= soft tissue growth) 
did not correspond to increase in length(= 
bone growth) as suggested by Matty (1985). 
No such depression was observed in an 
earlier study (Venkateshvaran et al. 1992). 
Fagerlund and McBride (1977) 
demonstrated statistically that the 
depression was not a direct effect ofhormone 
treatment but rather the natural effect of 
increasing size. 
v) Other considerations: 
There was no organised effort to study 
any possible side-effects such as renal 
hypertrophy or depression of the 
hepatosomatic index since these side effects 
are reported to manifest only at levels of 
8.3 ppm and above (Bulkley and Swihart, 
1973). 
It is customary to study the growth of 
fishes for varying periods upto 120 days or 
so subsequent to withdraw! of hormone 
5Days lODays 15Days 
1.00 1.10 0.98 
0.98 0.95 0.90 
1.01 0.91 0.88 
0.97 1.01 1.01 
1.00 0.95 0.97 
0.99 0.88 0.82 
administration. Such a study could not be 
carried out due to commercial disposal of 
fry, etc. 
As the fry are only to be restocked for 
further growing upto about 6 months, and 
are not meant for human consumption, the 
question of tissue residues was not looked 
into. 
ACKNOWLEDGMENTS 
The authors are thankful to Dr. G. 
Gopalkrishna, Scientist Incharge, 
Freshwater Fish Farm, Balabhadrapuram 
and Prof. K.M. Rao, Principal, CFETC, 
Kakinada for facilities provided and to Prof. 
Y. Sreekrishna, Director, CIFE, for 
permission to publish this paper. 
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Washington, D.C.: 1193 p. 
34 BORA, RAO AND VENKATESHVARAN 
Bulkley, R.V. and Swihart, G.L., 1973. 
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Field application of anabolic steroids in carp seed production. II. Rearing of spawn to fry stage.

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Field application of anabolic steroids in carp seed production. II. Rearing of spawn to fry stage.

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