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RESEARCH ARTICLE
Contents Vol 20(7)

Effect of density gradient centrifugation with trypsin on the in vivo fertilising capability of bovine spermatozoa

Brock A. Blevins A C , Morne de la Rey B and Naida M. Loskutoff A
+ Author Affiliations
- Author Affiliations

A The Bill and Berniece Grewcock Center for Conservation and Research, Henry Doorly Zoo, Omaha, NB 68107, USA.

B Embrio Plus, Brits, North West 0250, RSA.

C Corresponding author. Email: repro@omahazoo.com

Reproduction, Fertility and Development 20(7) 784-788 https://doi.org/10.1071/RD07197
Submitted: 25 October 2007  Accepted: 8 June 2008   Published: 1 August 2008

Abstract

The present study investigated the effect of a novel density gradient centrifugation (DGC) treatment using recombinant trypsin on the in vivo fertilising capability of bovine spermatozoa compared with a standard method. In Trial 1, semen collected from Boran and Ankole (Bos indicus) bulls was treated either with a silane-coated silica particle colloid formulated for humans with a recombinant trypsin or processed using a standard method (dilution in an egg yolk-based diluent). Semen processed by the two methods was used to artificially inseminate (AI) superovulated cattle. Day 7 embryos were flushed and assessed for fertilisation rates and embryo quality. Trial 2 used a trypsinised silane-coated silica particle colloid formulated specifically for bovine semen. Trial 1 resulted in significantly higher fertilisation rates using the trypsinised human DGC treatment than cows inseminated using the standard method (75.2% v. 67%, respectively; P < 0.01), but the numbers of transferable-quality Day 7 embryos did not differ between the two groups (P > 0.05). Results for Trial 2 indicated that cows inseminated with the trypsinised bovine DGC treatment had significantly increased fertilisation rates compared with the standard method (88.4% v. 63.1%, respectively; P < 0.01) and had significantly higher numbers of transferable-quality embryos (70.3% v. 51.8%, respectively; P < 0.01). In summary, bovine sperm treatment before AI by DGC and recombinant trypsin increases fertilisation rates and can result in more transferable-quality embryos compared with standard methods.

Additional keywords: embryo, recombinant trypsin, sperm processing, washing.


Acknowledgements

The authors thank Dr Paul Holmes (Nidacon International) for providing the PureSperm and BoviPure products used in this investigation. The authors also acknowledge Bernie Devlin for his assistance with data analysis. Special thanks to the veterinarians and hands at Ol Pejeta Ranch, Kenya.


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