Development of immune-reproductive biomarkers for improved breeding functionality in domesticated wolffish (Anarhichas minor)
1 Purpose: The experiment is dedicated to developing biomarkers related to the immune-reproductive mechanisms in the spotted wolffish broodstocks. Also, the hormonal administration for spawning to increase the production of gametes will be investigated.
2 Distress: Moderate distress will result as a consequence of repetitive management throughout the entire process. However, each time the animals are manipulated in a monthly basis, distress is minimal.
3 Expected benefit: (1) Optimize the existing short and long term sperm preservation protocols in wolffish, (2) develop an effective hormone administration regime to enhance the egg viability, quality, and quantity, (3) generate an immune-reproductive catalog of marker genes using biochemical and transcriptomic approaches, and (4) establish reliable welfare indicators for the domestication and management of the broodstock.
4 Number of animals, and what kind: The broodstock available is 48 spotted wolffish with an average age of 10 years each. For the experimental purposes proposed, The blood profile of eight animals will be tracked in an annual cycle to determine immune, nutritional, and biochemical changes as direct reporters on the regulation of the reproductive behavior. For the hormonal treatment, 30 animals divided into five individuals per treatment group will be used in the span of four weeks. This trial will be done twice within the 36 months requested experimentation.
5 How to adhere to 3R: We will follow the NORECOPA recommendations to improve fish welfare. Our experimental set-up minimizes the induction of stress (i.e., minimal handling and proper use of anesthetic) to the fish, and the personnel has adequate knowledge and training, especially in collecting biological samples (e.g., blood) and in the spawning induction (e. g., hormonal injection). Besides, we will use the minimum number of animals and treatments possible that will provide biological and statistical significance. Before performing any procedures, the fish will be anaesthetized using the commercial anesthetic (MS-222, Sigma-Aldrich) at a suitable dose of 500 ppm that has been previously optimized for this species. Moreover, the dissolved oxygen saturation level will be maintained >90% by continuous aeration, and the water temperature at 4–6°C by using the flow of the main pipe. The netting effort will be reduced only to hold the animals out of the tanks. After the procedures are finished, the fish will be returned immediately to their rearing environment and monitored continuously until their recovery.
- LeFrançois et al., 2021. Spotted Wolffish Broodstock Management and Egg Production: Retrospective, Current Status, and Research Priorities. Animals 11,2849.
- Superio et al., 2021. Spawning kinetics and parentage contribution of European sea bass (Dicentrarchus labrax) broodstocks, and influence of GnRHa-induced spawning. Aqua. Rep. 21:100766.
-Castejon et al., 2021. Vaccination of Gilthead Seabream After Continuous Xenoestrogen Oral Exposure Enhances the Gut Endobolome and Immune Status via GPER1. Front Immunol. 12:742827
*The full list of references is available on request.
2 Distress: Moderate distress will result as a consequence of repetitive management throughout the entire process. However, each time the animals are manipulated in a monthly basis, distress is minimal.
3 Expected benefit: (1) Optimize the existing short and long term sperm preservation protocols in wolffish, (2) develop an effective hormone administration regime to enhance the egg viability, quality, and quantity, (3) generate an immune-reproductive catalog of marker genes using biochemical and transcriptomic approaches, and (4) establish reliable welfare indicators for the domestication and management of the broodstock.
4 Number of animals, and what kind: The broodstock available is 48 spotted wolffish with an average age of 10 years each. For the experimental purposes proposed, The blood profile of eight animals will be tracked in an annual cycle to determine immune, nutritional, and biochemical changes as direct reporters on the regulation of the reproductive behavior. For the hormonal treatment, 30 animals divided into five individuals per treatment group will be used in the span of four weeks. This trial will be done twice within the 36 months requested experimentation.
5 How to adhere to 3R: We will follow the NORECOPA recommendations to improve fish welfare. Our experimental set-up minimizes the induction of stress (i.e., minimal handling and proper use of anesthetic) to the fish, and the personnel has adequate knowledge and training, especially in collecting biological samples (e.g., blood) and in the spawning induction (e. g., hormonal injection). Besides, we will use the minimum number of animals and treatments possible that will provide biological and statistical significance. Before performing any procedures, the fish will be anaesthetized using the commercial anesthetic (MS-222, Sigma-Aldrich) at a suitable dose of 500 ppm that has been previously optimized for this species. Moreover, the dissolved oxygen saturation level will be maintained >90% by continuous aeration, and the water temperature at 4–6°C by using the flow of the main pipe. The netting effort will be reduced only to hold the animals out of the tanks. After the procedures are finished, the fish will be returned immediately to their rearing environment and monitored continuously until their recovery.
- LeFrançois et al., 2021. Spotted Wolffish Broodstock Management and Egg Production: Retrospective, Current Status, and Research Priorities. Animals 11,2849.
- Superio et al., 2021. Spawning kinetics and parentage contribution of European sea bass (Dicentrarchus labrax) broodstocks, and influence of GnRHa-induced spawning. Aqua. Rep. 21:100766.
-Castejon et al., 2021. Vaccination of Gilthead Seabream After Continuous Xenoestrogen Oral Exposure Enhances the Gut Endobolome and Immune Status via GPER1. Front Immunol. 12:742827
*The full list of references is available on request.