Optimization of Nile Tilapia Artificial Breeding Using Human Chorionic Gonadotropin (hCG) Hormone
Abstract
1. Introduction
2. Experimental Design
Materials
- Mature male and mature female Nile tilapia (Oreochromis niloticus);
- Human chorionic gonadotropin (Merck, Darmstadt, Germany);
- NaCl (sodium chloride 98.80%) (Maldon, Essex, UK);
- Nitrate-Nitrite (0–40 mg/L NO3−-N, 0.015–0.60 mg/L NO2−-N) and ammonia test kit (0–2.4 mg/L NH3-N) (product numbers: 1408100 and 2428700, Hach, Loveland, CO, USA);
- 100 L plastic buckets;
- Heater (EHEIM thermocontrol 150 and 300 Watt, cat-4011708011713 and 4011708011744, Eheim GmbH & Co., Deizisau, Germany);
- Aerator (TetraTec APS 120-400, 400 L/h 4.5 watts, Tetra GmbH, Melle, Germany);
- Syringe (0.3 mL, Beckton-Dickinson, Oslo, Norway);
- Needle (Gauge 22, WVR, Radnor, PA, USA);
- Borosilicate glass beaker (500 mL);
- Aquarium tanks (500 L);
- Fine mesh net (7.5 cm)
- Lamp (Unilite SLR-3000, Worcestershire, UK);
- Timer (Bachmann, Bremen, Germany);
- Egg rocker (5 L capacity) (item number 1407, FIAP GmbH, Ursensollen, Germany);
- Air pump (Eheim Compacton 600 Pump, Deizisau, Germany);
- EBI temperature probe (item no. 620-1860P, WVR, Radnor, PA, USA);
- pH meter (item no. 665-0499P, WVR, Radnor, PA, USA);
- Oxygen meter (H05: Polaris C, Oxyguard, Farum, Denmark);
- Leica MZ16 FA dissecting stereomicroscope (Leica Microsystems GmbH, Wetzlar, Germany).
3. Procedure
3.1. Selection of Female Tilapia
3.2. Hormone Induction
3.3. Post-Injection Care
3.4. Selection of Male Tilapia
3.5. Egg Collection
3.6. Sperm Collection and Fertilization
3.7. Washing Fertilized Eggs
3.8. Incubation
3.9. Water Quality Parameters
3.10. Photoperiod
3.11. Monitoring the Fertilized Eggs
3.12. Data Collection
4. Results and Discussion
- Breeders should be maintained in a controlled wet lab environment for proper evaluation, ensuring the selection of superior females. Housing selected males and females in separate aquarium systems facilitates female maturation for spawning and allows more effective monitoring and management.
- Throughout the incubation process, we carefully monitored the eggs, removing any dead embryos to prevent deterioration of water quality.
- Sperm was stripped from the male and directly added to the eggs to ensure its viability. This immediate mixing helps maximize the chances of successful fertilization.
- It is essential to keep the eggs in motion at the bottom of the egg rockers to ensure successful development.
- The water in the tanks was changed once daily and replaced (75%) with freshwater manually using a bucket.
- When calculating the concentration of hormones used during breeding, particular attention should be given, as even slight variations in hormone dosage can have a significant impact on fertilization rates and larval survival. Proper dosing is crucial to ensuring successful reproduction and optimal outcomes in the breeding process.
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
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Group | Replicate | Total Eggs | Dead Eggs | Fertilized Eggs | Fertilization Rate (%) | Hatch-Out Larvae | Hatching Rate (%) | Total Surviving Larvae | Survival Rate (%) |
---|---|---|---|---|---|---|---|---|---|
32 °C | Rep 1 | 230 | 26 | 204 | 88.7 | 135 | 66.2 | 113 | 83.7 |
Rep 2 | 230 | 24 | 206 | 89.6 | 129 | 62.6 | 122 | 94.6 | |
Rep 3 | 230 | 20 | 210 | 91.3 | 127 | 60.5 | 116 | 91.3 | |
28 °C | Rep 1 | 230 | 22 | 208 | 90.4 | 140 | 67.3 | 128 | 91.4 |
Rep 2 | 230 | 32 | 198 | 86.1 | 145 | 73.2 | 131 | 90.3 | |
Rep 3 | 230 | 25 | 205 | 89.1 | 148 | 72.2 | 133 | 89.9 | |
24 °C | Rep 1 | 230 | 22 | 208 | 90.4 | 147 | 70.7 | 137 | 93.2 |
Rep 2 | 230 | 31 | 199 | 86.5 | 151 | 75.9 | 138 | 91.4 | |
Rep 3 | 230 | 40 | 190 | 82.6 | 153 | 80.5 | 136 | 88.9 |
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Rbbani, G.; Siriyappagouder, P.; Murshed, R.; Joshi, R.; Nedoluzhko, A.; Galindo-Villegas, J.; Fernandes, J.M.O. Optimization of Nile Tilapia Artificial Breeding Using Human Chorionic Gonadotropin (hCG) Hormone. Methods Protoc. 2025, 8, 57. https://doi.org/10.3390/mps8030057
Rbbani G, Siriyappagouder P, Murshed R, Joshi R, Nedoluzhko A, Galindo-Villegas J, Fernandes JMO. Optimization of Nile Tilapia Artificial Breeding Using Human Chorionic Gonadotropin (hCG) Hormone. Methods and Protocols. 2025; 8(3):57. https://doi.org/10.3390/mps8030057
Chicago/Turabian StyleRbbani, Golam, Prabhugouda Siriyappagouder, Riaz Murshed, Rajesh Joshi, Artem Nedoluzhko, Jorge Galindo-Villegas, and Jorge M. O. Fernandes. 2025. "Optimization of Nile Tilapia Artificial Breeding Using Human Chorionic Gonadotropin (hCG) Hormone" Methods and Protocols 8, no. 3: 57. https://doi.org/10.3390/mps8030057
APA StyleRbbani, G., Siriyappagouder, P., Murshed, R., Joshi, R., Nedoluzhko, A., Galindo-Villegas, J., & Fernandes, J. M. O. (2025). Optimization of Nile Tilapia Artificial Breeding Using Human Chorionic Gonadotropin (hCG) Hormone. Methods and Protocols, 8(3), 57. https://doi.org/10.3390/mps8030057