Fundamental cryobiology of selected African mammalian spermatozoa and its role in biodiversity preservation through the development of genome resource banking

Fundamental cryobiological characteristics of spermatozoa from threatened or endangered species must be determined for successful cryopreservation techniques to be established. In this study, spermatozoa from four diverse species, impala (Aepyceros melampus), wart hog (Phacochoerus aethiopicus), elephant (Loxodonta africana), and lion (Panthera leo), were collected by electroejaculation or epididymal aspiration. Spermatozoal plasma membrane permeability to water (hydraulic conductivity, L(p)) and the osmotically inactive fraction of the sperm cell (V(b)) were determined from each species. Changes in cell volume were measured over time using an electronic particle counter. A Kedem-Katchalsky membrane transport model was used to theoretically characterize the data to determine L(p) and V(b) for each species. In addition to determining plasma membrane characteristics, spermatozoa were also studied to determine their sensitivity to low temperatures and to permeating cryoprotectant solutes. Cells maintained at room temperature (20-22°C) were slowly or rapidly exposed to cold temperatures (1-4°C), and percent motility was estimated to determine the sensitivity of the cells to cooling. Spermatozoa were also in media containing 1 M glycerol, dimethyl sulfoxide or ethylene glycol, and percent motility was measured at 15, 30 and 60 min intervals to determine the sensitivity of the cells to the cryoprotectant agent over time. Results indicate that sperm motility is significantly effected by decreased temperatures and the presence of cryoprotectant agents.