Utility of carboxylated poly L-lysine for the liquid storage of bottlenose dolphin semen

Abstract

Assisted reproduction techniques are required to maintain a genetically diverse captive population of bottlenose dolphins. These techniques include semen preservation, and liquid storage has been proposed as a suitable alternative to cryopreservation, but the optimum conditions, in terms of temperature, duration, and media, have yet to be fully established. The aim of this study, therefore, was to determine the optimum temperature for the liquid storage of dolphin semen during a 14-day period and the usefulness of carboxylated poly-L-lysine (CPLL) as an additive to the semen extender used for the liquid storage. The semen was collected from a mature male dolphin housed at the Kagoshima Aquarium, Japan, transferred into a Beltsville (BF5F) extender, and analyzed for motility and characteristics after five-fold dilution. The optimum temperature was determined by evaluating sperm viability after liquid storage at 4, 17, or 36 °C, and the usefulness of CPLL was evaluated at concentrations of 0%, 0.5%, 1.0%, 2.0%, and 3.0% (v/v) at the optimum temperature. Sperm stored at 4 ℃ had a greater motility maintenance compared with samples stored at 17 or 36 ℃. The most efficacious storage regimen at various time points occurred when there was addition of CPLL at 1.0% (v/v) in terms of sperm motility and other relevant determinations, with this storage approach having greater efficacy that samples stored without CPLL. The most efficacious processes for preserving bottlenose dolphin sperm functions is storage at 4 °C and with there being semen extender supplementation of 1% CPLL.

Keywords: Bottlenose dolphin; Carboxylated poly-L-lysine; Cryopreservation; Semen extender; Semen storage.