Preliminary evidence on effect induced by HeiferPlus after in vitro exposure on functional parameters of buffalo's spermatozoa

Interest in buffalo breeding has largely increased worldwide as a dairy purpose animal. Since in a dairy farm heifer calves are significantly more valuable than bull calves, the birth of female animals prove to be an importantly aim. Recently, a new sexing agent (HeifersPlus™) for selecting X-chromosome bearing spermatozoa has been commercialized for the bovine. Up to now, no data are available on the use of HeifersPlus™ in the artificial insemination program in buffalo and on its influence on buffalo sperm quality after in vitro exposure. The aim of this study was to assess the influence of HeiferPlus™ treatment on some functional parameters of frozen-thawed buffalo spermatozoa. Semen of two Italian Mediterranean buffalo bull used in AI programme was used. Functional parameters assessed by cytofluorimetric analysis were: sperm viability (by using propidium iodide probe), acrosome reaction (by using PNA-FITC probe), sperm chromatin stability (by using acridine orange probe) and basal ROS determination (by using DCFH-DA probe). The treatment with HeiferPlus™ induced a significant (P≤0.05) reduction of sperm viability, whereas oxidative status and sperm chromatin stability were not affected by the treatment. Concerning acrosome reaction, the percentage of live spermatozoa at 0h and at 3h after thawing with and without Ca2+ exposure was significantly (P≤0.05) reduced after the treatment with HeifersPlus™. A significant reduction of reacted spermatozoa was observed at3h after exposure with HeifersPlus™ and Ca2+ treatment. From these data it seems that the treatment has adversely affected some of the functional parameter of buffalo spermatozoa after in vitro exposure.