Liquid storage of Ostrich (Struthio camelus) semen at 5 °C through intermediate dilution.

Dilution rate, dilution temperature and storage time have been recognized as vital steps in the processing of semen for storage before artificial insemination. The objective of this study was to determine optimal dilution and dilution temperature with an ostrich-specific semen extender for chilled storage. Four preselected ostrich (Struthio camelus var. domesticus) males, known for their ease of collection and specific semen quality parameters, were collected using the "dummy" female method. Dilution of 384 semen samples, at rates of 1:1, 1:2, 1:4 and 1:8 semen/diluent ratio with a diluent set at 5, 21 and 38 °C was performed and stored for 48 h at 5 °C. In vitro sperm function tests were conducted to evaluate treated semen during different storage intervals of 1, 5, 24 and 48 h. Motility and kinematic parameters were measured by the Sperm Class Analyzer®, the percentage live sperm measured by fluorescence SYBR14®/PI (LIVE/DEAD®), the percentage of sperm able to resist the hypo-osmotic swelling (HOS) stress test and sperm morphology determined by Nigrosin-Eosin staining. Progressive motility (PMOT), motility (MOT), sperm kinematics, LIVE and HOS were best (P < 0.05) maintained at a higher dilution of 1:4-1:8. The beneficial effect (P < 0.05) of a higher dilution temperature (21 °C) was prominent in terms of PMOT at a higher dilution. Storage of chilled semen at 5 °C requires dilution, at interpolated rates of 1:6-1:7, together with an extender temperature of 21 °C, to maintain optimal sperm function with minimal loss over a 48 h storage period.