CRISP protein expression in semen of the endangered Malayan tapir (Tapirus indicus)

The Malayan tapir is a large endangered herbivore native to South-east Asia with fewer than 2500 animals remaining in the wild. Although a small number of animals (183 animals held by 60 institutions) are managed in zoos and breeding centres, there is limited information on the fundamental reproductive biology of this species. The purpose of this present study was to evaluate the associations of reproductive protein biomarkers (CRISP2 and CRISP3) in the seminal plasma and spermatozoa with reproductive characteristics in male Malayan tapirs. Ejaculates were collected from zoo-housed animals by electroejaculation and assessed for sperm motility and quality traits. Seminal plasma and sperm pellets were analysed for CRISP protein expression by immunoblotting. The reproductive tract of a single animal was also analysed for CRISP2 and CRISP3 protein expression and localization by immunohistochemistry. Our results showed that both CRISP2 and CRISP3 are expressed in the seminal plasma and spermatozoa derived from Malayan tapirs. CRISP expression was positively correlated with semen quality, especially ejaculate volume, number of motile sperm, and acrosomal integrity. In addition, CRISP2 and CRISP3 protein expression were slightly high in males that had recently sired an offspring. The results suggest that CRISP proteins may serve as biomarkers for ejaculate quality and fertility in male Malayan tapirs. These findings may have significant implications for planning future breeding and re-introduction efforts for this species.