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Abstract
Nine groups each of four fish were injected with a single intramuscular dose of the following preparations: Physiological saline
(0.9% NaCl) as a control group, 0.5 ml kg1 Ovaprim, 20 and 40 g kg1 BW of GnRHa, 8 and 16 mL kg1 pimozide tablets
and the following combination of GnRHa with pimozide (GP): 20 g 4 mg, 30 g 8 mg and 40 g 16 mg kg1 BW.
The primary oocyte diameter (POD) before hormone administration ranged from 943.3 to 1071.0 m. The latency periods (LP)
were in the range of 9.0 to 12.0 h after injection. The highest ovulation ratio (OR) was observed in groups Ovaprim, GP(30
8) and GP(40 16). Other treatments were effective for ovulation, the ovulation ratio in Groups G(40) and GP(20 4) were
significantly higher than G(20) treatment. The ovulation index (OI) was in the range 62 to 77% and showed significant differences
among groups. There was no significant difference in fertilization ratio (FR) among Ovaprim, GP(30 8) and GP(40 16)
groups, while there were significant difference between the previous group and G(20) and G(40) groups. Control, P8, P16 showed
negative results in all the parameters LP, OED, OR, OI and FR. Levels of sex steroids were analyzed on 6 and 12 h after initiation
of treatments. A significant increase in plasma E2 with GP(30 8) injection was observed 6 and 12 h after injection, while there
were no significant increase between all the other groups 6 h after injection. Treatments with GP(20 4) resulted in a significant
increase in plasma T concentration in females compared with control after 6 h. In contrast, plasma T and E2 concentrations were
lower during the combined GP(20 4), GP(30 8) and GP(40 16) after 12 h than after 16 h of injection. The combined
treatments (GnRHa PIM) are better compared with Ovaprim which gave the same results, they have some advantages, such as
reliable response and low cost. Ovaprim is more than 3 to 5-fold of the cost of (GnRH PIM). Therefore, this method could be
useful tool for commercial catfish breeders to ensure spawning success.
2012 Elsevier Inc. All rights reserved.
Keywords: Clarias gariepinus; GnRHa; Pimozide; Ovaprim; Sex steroids; Ovulation
1. Introduction
In nature, African catfish, Clarias gariepinus, has a
discontinuous annual reproductive cycle with alternate
periods of resting, pre-spawning and breeding, regulated by cyclically active gonadotrophes [1]. The breeding season correlates with periods of maximal rainfall
* Corresponding author. Tel.: 201224173750.
E-mail address: safaa_sharaf@agr.suez.edu.eg (S.M. Sharaf).
0093-691X/$ see front matter 2012 Elsevier Inc. All rights reserved.
doi:10.1016/j.theriogenology.2011.12.019
1710
Table 1
Substances and doses applied to stimulate ovulation in Clarias
gariepinus females (n 4).
Treatment groups
Control
Ovaprim
G(20)
G(40)
P8
P16
GP(20 4)
GP(30 8)
GP(40 16)
Substance
Dosage
Saline (0.9%NaCl)
Ovaprim
GnRHa
GnRHa
PIM
PIM
GnRH PIM
GnRH PIM
GnRH PIM
20 kg1
40 kg1
8 mg
16 mg
20 kg1 4 mg
30 kg1 8 mg
40 kg1 16 mg
sies were taken, only fish having more than 60% of the
oocytes with a migrating germinal vesicle were selected
for ovulation experiment. Thirty-six female fish weighing 500 to 1000-g body weight (BW) were selected.
Before injection fish were individually weighed and
assigned to nine groups. No food was provided during
the experiment.
2.2. Hormones and experimental design
Nine groups of four fish each were injected a single
intramuscular dose into the dorsal muscle above lateral
line with different preparations as follows: Physiological saline (0.9% NaCl) was injected as control group,
Ovaprim (Syndel Laboratory, Limited, Canada) 0.5 ml
kg1 which contains the synthetic GnRH analog and
domperidone, 20 and 40 g kg1 BW of GnRH {42 g
kg1 buserelin acetate (Pyr-HisTrpSerTyr-D-Ser(But)-LeuArgPro ethylamide) and 10 mg benzyl alcohol), synthetic, Receptal, Intervet, Germany}, 8, 16
ml kg1 pimozide tablets (Orap Forte, Cilag, Belgium)
were powdered and then dissolved in physiological
saline and dimethyl sulfoxide, respectively [33] and the
following combination of GnRH with pimozide: 20 g 4
mg, 30 g 8 mg and 40 g 16 mg kg1 BW
(Table 1).
After injection fish were placed in well-aerated tanks
with recirculated water and temperature of 28 1 C.
The first examination for ovulation was carried out 6 h
after injection and repeated every hour. So when ovulation was observed, the eggs were stripped manually
after the latency period (mean time between injection
and ovulation [23]) and weighed from each female
separately, the weight being expressed as percentage of
female BW. The number of eggs released was calculated following the gravimetric method [34]. Assessment of ovulation was carried out by determining the
ovulation success (number of ovulated females/number
of injected) 100% and by ovulation index (OI) weight
1711
1712
Table 2
The effect of different treatments on latency period (h), ovulatory egg diameter (mm), ovulation ratio (%), ovulation index (%), Fertilization
success (%) and hormone cost of Clarias gariepinus.
Treatment
groups
Primary oocyte
diameter (m)
Latency
period (h)
Ovulated egg
diameter (m)
Ovulation
ratio%
Ovulation
index%
Fertilization
ratio%
Cost a (LE)
Control
Ovaprim
G[20]
G[40]
P8
P16
GP(20 4)
GP(30 8)
GP(40 16)
836.93 147.08
929.33 64.0
727.63 49.01
780.47 87.17
708.93 63.62
677.10 73.04
710.40 90.21
895.77 72.16
682.13 54.04
986.67 24.04
1033.3 44.1
1071.0 72.3
1026.7 89.7
966.7 60.1
973.3 99.6
943.3 64.4
1010.0 51.3
983.3 44.1
0
9.9c 0.23
11.6a 0.2
11.5a 0.24
0
0
11ab 0.36
10.5bc 0.37
11ab 0.41
0
1140.9ab 64.3
1050.5b 28.8
1190.5ab 21.9
0
0
1154.0ab 55.0
1154.2ab 39.1
1239.9a 115.9
0
100a
20d
60c
0
0
90b
98a
100a
0
73.1a 1.6
62.1b 1.2
66.8b 1.8
0
0
62.2b 2.2
73.4a 2.6
77.1a 2.5
0
84.0 0.6
77.3 2.8
79.3 1.2
0
0
81.0 1.5
81.5 0.6
83 1.7
0
30.0
2.5
5.0
2.0
4.0
3.5
5.75
9.0
The same superscript in the same column is not significantly different at P 0.05.
a
$1.0 LE 6.0.
4. Discussion
The present study showed the effect of administration of synthetic hypothalamus hormones combined
with the antidopaminergic antagonist PIM on spawning
in C. gariepinus. The failure of fish to ovulate after
treatment with GnRHa or PIM and the control vehicles
may suggest that plasma gonadotropin (Gth) levels of
those fish had remained low. A surge in Gth can initiate
final events of oocyte maturation and ovulation in C.
macrocephalus, as in C. gariepinus [42]. In other fish,
administration of LHRHa or PIM resulted in higher Gth
levels than the controls, but the ability to cause oocyte
maturation and ovulation depended on the dose, species
[14,17,19] and the maturational stage of the oocytes
14
C
va
O
on
tr
ol
d
C
cd
cd
C
)
P(
40
+1
6)
P(
30
+8
G
P(
20
+4
P(
16
)
P(
8)
G
(4
0)
G
(2
0)
va
B
cd
)
P(
40
+1
6)
bcd
cd
P(
30
+8
abcd
AB
abc
bc
bcd
A
P(
20
+4
bcd
P(
16
)
cd
abcd
B
AB
ab
P(
8)
AB
AB
G
(4
0)
AB a
G
(2
0)
AB
12 h
10
T (ng/L)
12 h
6h
12
6h
10
9
8
7
6
5
4
3
2
1
0
on
tr
ol
E2 (ng/L)
Treatments
Treatments
1713
1714
[9]
[10]
[11]
[12]
[13]
Acknowledgments
The author acknowledges all the members in Animal
Production and Fish Resources Department and Fish
Research Center, Suez Canal University, Ismailia,
Egypt for their assistance in fish husbandry and sampling. The author also thanks the central laboratory for
aquaculture research Abbassa, Abo-Hammad, Sharkia,
Egypt.
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