Assessment of basic seminal characteristics, sperm cryopreservation and heterologous in vitro fertilisation in the fishing cat (Prionailurus viverrinus)

Abstract

Conservation of the fishing cat, a threatened south-east Asian felid, could benefit from effective ex situ genetic management and breeding programmes, including the use of assisted reproduction. The aims of the present study were to: (1) characterise basal seminal traits of fishing cats in Thailand zoos; and (2) investigate the effect of cryopreservation on sperm motility, acrosomal integrity and in vitro function. Seminal traits were evaluated in electroejaculates collected from eight males. Spermatozoa were diluted in n-tris(hydroxymethyl)-methyl-2-aminoethanesulfonic acid Tris (TEST)-yolk buffer (TYB) without glycerol, then diluted further with TYB with glycerol (4% final concentration) at either 25 degrees C or after slow cooling to 5 degrees C and frozen in straws over liquid nitrogen vapour. After thawing, sperm function was assessed by insemination of viable domestic cat oocytes. Fishing cat ejaculates averaged (+/- s.e.m.) 43.6 +/- 14.2 x 10(6) motile spermatozoa with 33.5 +/- 6.8% normal sperm morphology. Semen processing had a negligible effect (P > 0.05) on sperm motility and acrosomal integrity, but values were reduced (P < 0.05) after thawing. All thawed samples fertilised domestic cat oocytes, with 62.1% (36/58) of mature oocytes cleaving. Glycerol addition at 5 degrees C resulted in higher (P < 0.05) post-thaw motility and intact acrosomes than glycerol addition at 25 degrees C. In conclusion, good-quality ejaculates can be obtained from Thai fishing cats and their spermatozoa exhibit adequate function after cryopreservation for in vitro fertilisation procedures.

Cryopreservation of feline epididymal spermatozoa from dead and alive animals and its use in assisted reproduction

“…This fact is probably relevant also in felids, as in the fishing cat (Prionailurus viverrinus) postthaw motility and acrosome integrity were higher after glycerol addition at 5 • C than at 25 • C (Thiangtum et al, 2006).…”

Sperm cryopreservation in the Far‐Eastern wildcat (Prionailurus bengalensis euptilurus)

“…The common methods to test the viability of spermatozoa after freezing–thawing are fluorescent and light microscopy accompanied with staining; estimation of sperm concentration, the percentages of live and dead sperms, and sperm motility; and evaluation of sperm morphology (Amstislavsky et al, ; Ha et al, ). The ultimate test for semen fertilizing capacity is IVF followed by IVC (Kunkitti, Axnér, Bergqvist, & Sjunnesson, ; Thiangtum et al, ).…”

“…Along with the Far-Eastern wildcat, the genus Prionailurus includes such endangered species as the fishing cat (Prionailurus viverrinus) (Thiangtum et al, 2006) and the flat-headed cat (Prionailurus planiceps) (Wilting et al, 2010). At present, the sperm of these two species has been successfully frozen (Thiangtum et al, 2006;Thuwanut et al, 2011). With the only exception of Ha et al (2011), who cryopreserved spermatozoa of two leopard cat males of unknown subspecies, there were no attempts to freeze sperm of the Far-Eastern wildcat.…”

In vitro growth and steroidogenesis of dog follicles are influenced by the physical and hormonal microenvironment

“…For example, we could envision laboratories dedicated to rescuing follicles from ovaries removed expeditiously from valuable animals that die unexpectedly or undergo ovarioectomies for medical reasons. As sperm cryopreservation has become routine for many mammals, including a diversity of carnivores (Goodrowe et al 2000, Crosier et al 2006, Thiangtum et al 2006, Asa et al 2007), viable oocytes recovered from cultured follicles could be used to generate embryos in vitro . This capacity would enhance the management of precious genotypes representing dogs used as models for studying human disorders (Schneider et al 2008) or the conservation of a growing list of wild canid species threatened in nature (IUCN 2010).…”