Insights on sperm assays and cryopreservation in six Neotropical pit vipers

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dc.contributorLab. Herpetologiapt_BR
dc.contributor.authorBlank, M.H.pt_BR
dc.contributor.authorNovaes, G.A.pt_BR
dc.contributor.authorLosano, J.D.A.pt_BR
dc.contributor.authorSant'Anna, Sávio Stefaninipt_BR
dc.contributor.authorVieira, Samira Emanuela Mariapt_BR
dc.contributor.authorGrego, Kathleen Fernandespt_BR
dc.contributor.authorPereira, R.J.G.pt_BR
dc.identifier.citationBlank M.H., Novaes G.A., Losano J.D.A., Sant'Anna SS, Vieira SEM, Grego KF, et al. Insights on sperm assays and cryopreservation in six Neotropical pit vipers. Cryobiology. 2022 June;106:55-65. doi:10.1016/j.cryobiol.2022.04.003.pt_BR
dc.description.abstractBreeding snakes in captivity has become more and more relevant due not only to the growing interest on their venoms but also to the increasing number of endangered species worldwide. Unfortunately, studies on the formation of germplasm banks for these reptiles do not follow the same pace, and literature on sperm cryopreservation remains in its infancy when compared to other taxa. Herein, we first validated a sperm-egg binding assay (using chicken egg perivitelline membrane – EPM) and some nonfluorescent staining techniques for semen analysis of two pit viper genera (Bothrops and Crotalus), and then we investigated the protective effects of dimethylacetamide (DMA), dimethylformamide (DMF), and dimethylsulfoxide (DMSO) at different concentrations (3, 6 and 12%) throughout the freezing process in five species of lancehead and one of rattlesnake. Our validation process showed high correlations among sperm functional tests (including sperm-binding to EPM) and motion parameters. A total of 166 fresh ejaculates were acquired from 233 collection attempts, and 63.9% of these samples exhibited minimal motility for freezing (≥20%). During cryopreservation we observed that post-thaw motility and quality was improved by higher levels of cryoprotectants (CPA), regardless the CPA type. Lower concentrations of CPA were less harmful to sperm motility and progressive motility following the equilibrium phase, but were ineffective in protecting these cells from the freeze-thaw cycle. Likewise, higher CPA concentrations increased post-thaw integrity of the acrosome and plasma membrane for most species, except for rattlesnakes in which only 12% DMSO produced better outcomes.pt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.description.sponsorship(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorpt_BR
dc.rightsRestricted accesspt_BR
dc.titleInsights on sperm assays and cryopreservation in six Neotropical pit viperspt_BR
dc.contributor.external(USP) Universidade de São Paulopt_BR
dc.subject.keywordreproductive technologiespt_BR
dc.subject.keywordsemen analysispt_BR
dc.subject.keywordsperm freezingpt_BR
dc.identifier.citationabntv. 106, p. 55-65, jun. 2022pt_BR
dc.identifier.citationvancouver2022 June;106:55-65pt_BR
dc.contributor.butantanSant'anna, Sávio Stefanini|:Pesquisador|:Lab. Herpetologiapt_BR
dc.contributor.butantanVieira, Samira Emanuela Maria|:Técnico|:Lab. Herpetologiapt_BR
dc.contributor.butantanGrego, Kathleen Fernandes|:Pesquisador|:Lab. Herpetologiapt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2012/08621-1pt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2016/17,158-4pt_BR
dc.sponsorship.butantan(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior¦¦pt_BR
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