Study of fertilization in fish through induced spawning under
laboratory conditions.
Induced spawning in fish:
Induced spawning is one of the common methods to stimulate ovulation of the fish in the hatchery. With this method, hormones
are playing an important role. Several natural and artificial hormones have been used to induce breeding of fish, while Ovaprim is
one of the most popular and effective solutions to stimulate the maturation of male and female broodfish. Ovaprim contains
combinations of salmon gonadotropin-releasing hormone (sGnRH) and domperidone. These hormones have been applied
successfully to induce spawning of fish.
Method
Study timeline and site
The experiments were carried out within the ethical guidelines in animal research. The study was conducted between April and
June 2019, at the Fish Breeding Laboratory, University of Education, Lahore. The broodfish ranged between 10 and 15 cm in
total length, and 18.42 and 31.15g in total weight. The fish were acclimatized for 24 hours prior to inducing with Ovaprim.
Hormone administration
The experiment was carried out in compliance with the ethical guidelines. Three dosages of Ovaprim were tested in this study:
for females, 0.3 ml kg-1 body weight (BW), 0.5 ml kg-1BW, and 0.7 ml kg-1BW; for males, 0.15 ml kg -1BW. The broodfish in
control groups were injected with physiological solution (0.9% NaCl) at the dosage of 0.25 ml kg -1BW.
Sperm and egg collection
The female broodfish were injected with their respective dosage two times; the first injection was at 8.00 PM with half of the
tested dosage, and the second injection was conducted 6 hours after the first injection (at 2.00 AM), with the remainder of the
tested dosage. The female broodfish was ovulated 6 hours after the second injection (at 8.00 AM). The eggs were collected by
gentle finger pressure to the abdomen. The collected eggs were put in a plastic jar and kept in an ice box at 4°C 6.
The males were injected with a single dose of 0.25 ml kg -1 BW at 8.00 PM. The male fish was sacrificed. The testes were removed
and washed with physiological solution and then perforated and chopped with scissors. The semen was gently squeezed out and
put in a tube, and kept in an icebox (4°C). The semen was mixed with physiological solution (0.9% NaCl) at dilution ratio 1: 100
(v/v).
Fertilization and incubation
A total of 1 ml eggs and 1 ml of diluted sperm were mixed homogenously in a plastic basin developed by the Laboratory of Fish.
The resulting mixture was left in contact for 5 minutes. The incubation basin was installed with 24-hour portable aerator at a
water temperature of 25–27°C in a water heater. Successful fertilization was observed 8 hours after incubation. Unfertilized eggs
were identified by their opacity; the unfertilized eggs were removed from the jar, while hatching rate was monitored at two-hour
intervals. The larvae were fed on day 5 after being hatched and reared in the same jar for 40 days.
Measured parameters and data analysis
The latency period, the relative number of ovulated eggs, egg diameter, fertilization rate, hatching rate, and survival rate of larvae
on day 40 after hatching were measured. The latency period was determined by calculating the time between the second injection
and ovulation. The relative number of ovulated eggs was calculated by dividing the total number of released eggs after Ovaprim
injection by the total body weight of the female broodfish.
Results:
The result showed that the latency period was faster at a dosage of 7 ml kg -1BW; this value was significantly different from other
dosages. A higher relative number of ovulated eggs and fertilization, hatching, and survival rates were also recorded at the
Ovaprim dosage 7 ml kg -1 BW; these values are significantly different from other dosages. In general, the relative number of
ovulated eggs, fertilization, hatching, and survival rates were increased with increasing Ovaprim dosage. However, there were no
significant differences between the values seen at 3 ml kg -1 BW and 5 ml kg-1 BW dosage.
Discussion
The results indicate a decreased latency period at increased Ovaprim dosage. On the contrary, the relative number of ovulated
eggs, fertilization, hatching, and survival rates were increased at increased dosage. The best results for all parameters were
recorded at the Ovaprim dosage of 7 ml kg -1 body weight. Therefore, the higher dosage of Ovaprim (7 ml kg -1 BW) was an
effective dosage in inducing spawning of the fish compared to the lower dosage (3 ml kg -1 and 5 ml kg-1). This is probably
because the combination of sGnRH and domperidon in Ovaprim solution at higher doses of 7 ml kg -1 BW stimulated
gonadotropin (GTH II or LH) secretion in the pituitary gland of the broodfish to a greater extent, and then induced gonad
maturation and ovulation. The higher levels of gonadotropin will induce ovulation faster . Besides being influenced by hormonal
factors, the hatching rate is also influenced by the temperature of the incubation media, dissolved oxygen, pH, and light intensity.
Conclusions
It is concluded that different doses of Ovaprim had a significant effect on the latency period, the relative number of ovulated
eggs, fertilization, hatching, and survival rates of fish.
