250 DEHYDRATED COCONUT WATER FOR IN VITRO SPERM CAPACITATION IN SWINE
A.B. Nascimento A B , V.P. Oliveira B , M.G. Marques B , R. Toniolli A , R.P.C. Gerger B , A.R.S. Coutinho B , W.B. Feitosa B and J.A. Visintin BA Swine Reproduction and Sperm Technology Laboratory – Federal University of Ceara (UECE), Foraleza, Brazil
B Department of Animal Reproduction – University of Sao Paulo (USP), Sao Paulo, Brazil. Email: aballarotti@yahoo.com.br
Reproduction, Fertility and Development 17(2) 275-275 https://doi.org/10.1071/RDv17n2Ab250
Submitted: 1 August 2004 Accepted: 1 October 2004 Published: 1 January 2005
Abstract
Recently, progress has been achieved in reproduction biotechnology with coconut water as semen extender for domestic animals and in bovine oocyte in vitro maturation and embryo culture. The aim of this study was to evaluate the dehydrated coconut water (ACP250) as semen extender and its effect on in vitro sperm capacitation in swine. Eighteen ejaculates were collected from three crossbred boars of Landrace and Duroc. Semen samples were extended in ACP250 solution or Beltsville Thawing Solution (BTS) and submitted to sperm capacitation after extension (EXT) or after cooling (COO) at 16°C for 24 h. The semen was centrifuged at 400g for 8 min. After centrifugation, the samples were standardized at 2 × 107 spermatozoa/mL, subjected to sperm capacitation in TALP medium with 5 mM of caffeine, and incubated at 38.5°C and 5% of CO2 in high humidity for three h. Four treatments were tested, T1 (ACP250 + EXT), T2 (ACP250 + COO), T3 (BTS + EXT), and T4 (BTS + COO). Sperm capacitation was evaluated by Coomassie Blue G stain, as follows: capacitated spermatozoa were stained in tenuous blue and not capacitated in intense blue. The Coomassie Blue G staining technique was described by Larson and Miller (1999 Mol. Reprod. Dev. 52, 445–449), who demonstrated that sperm from a variety of mammalian species can be stained by this technique, and an adequate observation of the acrosomal status was possible as well. This procedure was described as simple, fast, inexpensive and reliable, and it requires no fluorescence or DIC optics. For statistical analysis, SAS (System for Windows; SAS Institute, Inc., Cary, NC, USA) was utilized (P < 0.05). Comparing the sperm capacitation rates in relation to fresh vs. cooled semen, there were significant differences between T1 (22.3%) and T2 (43.7%) and between T3 (23.4%) and T4 (48.1%) (P < 0.05). In relation to BTS or ACP250 extenders, there were no significant differences between T1 and T3 and between T2 and T4 (P > 0.05). In conclusion, the ACP250 solution is an option for in vitro sperm capacitation in swine, specially with chilled extended semen.
This work was supported by FAPESP 01/11931-8 and ACP Biotechnology – CE.
