Validation of advanced tools to evaluate sperm function in barramundi (Lates calcarifer).

Abstract Barramundi (Lates calcarifer) is a tropical finfish species rapidly growing in popularity for aquaculture production. However, sperm quality tests have yet to be adapted to enable selection of highly fertile male broodstock in this species. Accordingly, in this study advanced tools were optimized to evaluate barramundi sperm function to facilitate the future study of male fertility and address some of the reproductive constraints currently observed in captive-bred broodstock. Sperm morphology data were used to calibrate and validate automated sperm counting and motility detection by computer-assisted sperm analysis (CASA; AndroVision, Minitube). Several parameters were examined to determine the optimum settings for accurate CASA sperm counting and were compared to manual haemocytometer methods including: sample dilution (1:1000, r = 0.87), minimum number of fields (n = 4, CV = 7.5%), and the effect of motile vs. immotile spermatozoa on automated counting (no effect, r = 0.99, P