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Artificial insemination of bitches with sexed semen by density gradient

Abstract

Different techniques are described for semen sexing in bovine. The semen centrifugation in discontinuous density gradient method is described as effective in some species, although it has questionable accuracy in most cases. However, this method has reduced costs when compared to flow cytometry. In dogs, there are no studies about sexing sperm in the literature, despite the demand by dog breeders for a particular sex that favors the sale of animals and the high growth of the pet industry. Therefore, this study aims to study a technique compatible with the reality of dog breeders, affordably and without the need for sophisticated equipment. The study will be divided into two stages, the Experiment I will evaluate between three protocols of density gradient centrifugation (Percoll, Nycoprep associated with Percoll and Ficoll). Sperm cells are evaluated pre- and post-centrifugation for motility by CASA (Computer-Aided Sperm Analyzer), concentration of the ejaculate, sperm morphology, plasma and acrosomal membrane integrity, and mitochondrial function. The samples before and after centrifugation will be evaluated by RTqPCR to identify the method that can separate increased number of sperm containing the X chromosome. After determining the most appropriate method, Experiment II will test the in vivo fertility of sperm cells separated by the method which separated much sperm X, using artificial insemination. It is expected to get more females and litter size compatible with the race when the results are compared to control. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MOTHE, GABRIELE BARROS; SCOTT, CAROLINE; SICHERLE, CARMEN CECILIA; DE FREITAS GUAITOLINI, CARLOS RENATO; FREITAS DELL'AQUA, CAMILA DE PAULA; MALOSSI, CAMILA DANTAS; ARAUJO-JUNIOR, JOAO PESSOA; DE SOUZA, FABIANA FERREIRA. Sperm sexing with density gradient centrifugation in dogs. Animal Reproduction Science, v. 199, p. 84-92, . (12/20670-8)

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