Effect of antibiotic supplementation on the quality of cryopreserved fish sperm of silver barb (Barbodes gonionotus): Sperm motility and viability, bacterial quality and fertilization

Abstract

Disease prevention is a key aspect in developing cryopreservation procedures for fish sperm and in improving the reproductive biotechnology for commercially important aquatic species. The purpose of this study was to evaluate the effects of an antibiotic supplementation (0.25% penicillin–streptomycin, PS and 0.25% penicillin–gentamicin, PG) on sperm motility and viability, bacterial profile and fertilization capacity of cryopreserved silver barb (Barbodes gonionotus) semen. The experimental protocol involved three treatments: addition of PS alone; addition of PG alone; and no addition of antibiotics (Control). Semen samples were frozen and cryostored for 12-mo. Administration of 0.25% PS significantly (P<0.05) improved sperm motility and viability and reduced (P<0.05) total heterotrophic bacteria, Gram negative bacteria and pseudomonads bacteria in cryopreserved semen. Post-thawed semen treated with 0.25% PS did not contain contaminating bacteria including Bacillus subtilis subsp. inaquosorum, Bacillus safensis, Aeromonas punctata subsp. caviae, Serratia plymuthica, Pseudomonas azotoformans and Pseudomonas sp. Post-thawed semen supplemented with 0.25% PG showed degenerative changes in motility and viability of sperm. Eggs fertilized with 0.25% PS or antibiotic-free cryopreserved semen had similar fertilization rates, lower (P<0.05) than those of fresh semen. Incorporation of 0.25% PS was suitable for cryopreservation of silver barb semen based on the presence of good quality of post-thawed sperm and elimination of bacterial contaminants: B. subtilis subsp. inaquosorum, B. safensis, A. punctata subsp. caviae, Ser. plymuthica, P. azotoformans and Pseudomonas sp. This is the first study to investigate the antibiotic effect on the number of bacteria and their profile in cryopreserved semen of fish.