Preservation of Black Tiger Shrimp Penaeus monodon Spermatophores by Chilled Storage

Chilled storage of spermatozoa in fish has been extensively investigated for many years, but limited research was focused on crustacean species. Chilled storage of spermatophores of black tiger shrimp Penaeus monodon is needed to generate consistent and reliable supply of spermatozoa for subsequent use. The objective of this study was to develop a protocol for the chilled storage of black tiger shrimp spermatophores and to evaluate bacterial propagation during chilled storage of spermatophores. In the first experiment, spermatophores were selected and preserved using four different extenders, namely mineral oil, Ringer's solution, phosphate buffer and 0.85% sodium chloride, and stored at low temperature (2‐4 C) for 42 d without antibiotic supplementation. Results showed that mineral oil was the best extender for chilled storage of spermatophores, since the highest percentage of viable sperm (58.3 ± 2.9%) was observed with this extender at the end of experiment (day 42). Bacillus sp., Staphylococcus sp., and Pseudomonas aeruginosa were identified as the predominant bacteria occurring during chilled storage, and the total bacteria count gradually increased during the experiment. In the second experiment, spermatophores were preserved in the mineral oil with four concentrations of the antibiotic, penicillin‐streptomycin (0.1%,1%, 2%, and 3%). There was no significant difference (P 0.05) in the percentage of viable sperm among treatments with 0.1%,1 %, 2%, and 3% antibiotics. The total count of Bacillus sp., Staphylococcus sp., and P. aeruginosa in the antibiotic treated groups significantly decreased (P < 0.05) to undetectable levels by day 14 of the experiment. Fertility studies from artificial insemination indicated that P. monodon spermatophores preserved with mineral oil for 7‐8 d at 2‐4 C were capable of fertilizing eggs with hatching rates similar to the controls. This study suggests that chilled storage of spermatophores is a feasible approach for the management and spawning of black tiger prawn broodstock or other Invertebrate species that produce spermatophores.