Two semen extenders were compared for their ability to maintain viability of horse semen during 24 hours of cold preservation, and for the pregnancy rate after artificial insemination. In the experiment 1, five ejaculates from three stallions were split-diluted in either a skimmed milk-based extender (Kenney extender) or a glycine egg yolk-based extender (Foote extender) and cooled at 6-8 ºC for 24 hours. Semen samples stored in Kenney extender for 24 hours had higher motility and spermatic vigor compared with those stored in Foote extender. However, samples stored in Foote extender had higher number of reactive sperm by hypoosmotic test and greater viability by epifluorescence test compared with those in Kenney extender. In the experiment 2, 17 and 23 ejaculates from two stallions were split-diluted with Kenney extender and Foote extender. The sperm concentration in each extender was adjusted to 500 million viable sperms per insemination dose. Semen was cooled to 6-8 ºC and stored for 24 hours. Seventy-four cycles of crossbred mares were inseminated with either semen diluted in Kenney extender or semen diluted in Foote extender. The pregnancy rate was higher from semen diluted in Kenney extender than that from semen in Foote extender (0.553 vs. 0.306). The Kenney extender is effective in preserving the motility, vigor and fertility of stallion semen after 24 hours of cold storage, whereas the Foote extender is not acceptable.

O objetivo neste estudo foi avaliar diferentes osmolaridades e tempos de incubação durante o teste hiposmótico e suas correlações com a congelabilidade do sêmen de touros zebuínos. Foram utilizados 30 ejaculados de seis touros adultos da raça Nelore. No sêmen in natura foi feita avaliação física e morfológica, classificação quanto à coloração supravital e o teste hiposmótico. No teste hiposmótico foram utilizadas soluções com osmolaridades de 60, 100, 150 mOsm/kg e água destililada (19 mOsm/kg) om 15, 30 e 60 minutos de período de incubação a 37ºC. Após a criopreservação, as amostras foram descongeladas e avaliadas quanto aos testes hiposmótico, à coloração supravital, ao teste de termorresistência lento e à coloração fluorescente. Não foram detectadas interações entre os tempos e as osmolaridades. Não houve diferenças para os valores médios de espermatozoides reativos incubados nas soluções com diferentes osmolaridades para o sêmen congelado/ descongelado. Apenas a utilização de água destilada determinou diferença nos valores médios obtidos no teste hiposmótico realizado no sêmen in natura. Não houve diferença entre os valores médios de espermatozoides reativos incubados com diferentes tempos de incubação, tanto no sêmen in natura como no sêmen congelado/descongelado. O teste hiposmótico pode ser realizado com 15 minutos de incubação, e com uma solução que varie a osmolaridade de 60 a 150 mOsm/kg, tanto no sêmen in natura como no sêmen congelado/descongelado.

The objective of this study was to evaluate different osmolarities and incubation times during the hiposmotic test and its correlation with cryopreservation of semen of Zebu bulls. It was used 30 ejaculates from six mature Nelore bulls. It was performed physical and morphological evaluation, supravital staining classification and hiposmotic test in the fresh semen. In the hiposmotic test, solutions with osmolarities of 60, 100, 150 mOsm/kg and distilled water (19 mOsm/kg) were used at 15, 30 and 60 minutes of incubation at 37ºC. After cryopreservation, the samples were thawed and evaluated for hiposmotic test, supravital staining, slow termoresistance test and for fluorescent stain. No interactions were detected between incubation times and solution osmolarities. There were no differences in average values of reactive sperm incubated in different solutions with different osmolarities for frozen/thawed semen. Only the use of distilled water determined differences in the average values obtained in the hiposmotic test performed in fresh semen. There was no difference between the mean values of reactive spermatozoa incubated with different incubation time in both fresh and frozen/thawed semen. Hiposmotic test can be accomplished in 15 minutes of incubation time and with a solution which varies osmolarity from 60 to 150 mOsm/kg in both fresh and frozen/thawed semen.