Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Semina. Ciências Agrárias (Online) |
DOI: | 10.5433/1679-0359.2016v37n3p1643 |
Texto Completo: | https://ojs.uel.br/revistas/uel/index.php/semagrarias/article/view/22003 |
Resumo: | Prochilodus brevis is a migratory fish, an important component of its river ecosystem and an appreciated animal in northeastern cuisine. However, human activities have threatened its survival. Thus, researchers have become interested in developing genetic material storage protocols, such as seminal cryopreservation. Therefore, determination of the appropriate freezing media and thawing rate is a fundamental step toward the use of this biotechnology in the production of common curimatã and for reducing risks to the species’ survival. As such, the aim of the current study was to evaluate the effect of different freezing media and thawing rates on the quality of cryopreserved semen from P. brevis. For this study, males received a single dose of pituitary extract of carp 18 hours before semen collection. The semen samples were diluted in 5% glucose + 10% methyl glycol (MG), 5% glucose + 10% dimethyl sulfoxide (DMSO), 0.9% NaCl + 10% methyl glycol, and 0.9% NaCl + 10% dimethyl sulfoxide, loaded into 0.25-mL straws and frozen in liquid nitrogen vapor. The semen from each treatment was thawed at three different thawing rates: 25 °C for 30 sec, 30 °C for 16 sec and 40 °C for 12 sec. Motility, vitality and morphology analyses were performed by computer-assisted sperm analysis (CASA). The characteristics of the fresh sperm mostly resembled those found in the literature. For the parameters analyzed, fresh sperm presented higher sperm quality in comparison to all treatments with cryopreserved sperm (p < 0.05), except for the characteristic of normal morphology, for which the sperm cryopreserved in glucose and MG did not differ statistically from the fresh sperm. For the cryopreserved semen, the greatest results of total motility and curvilinear velocity (VCL) were obtained using glucose and DMSO, regardless of the thawing rate employed. For the straight-line velocity (VSL) and average path velocity (VAP), DMSO showed the best results, regardless of the diluent and thawing rate. With regard to vitality, the highest values were achieved when DMSO and thawing rates of 30 °C for 16 sec or 40 °C for 12 sec were used. In the morphological analysis, the greatest percentage of normal sperm cells was obtained using thawing rates of 25 °C for 30 sec and 40 °C for 12 sec, regardless of the freezing media. Sperm quality was found to suffer interference from the freezing media, as well as from interaction between its components (diluent and cryoprotectant) and the thawing rate used. Under the methodological conditions employed, the use of 5% glucose + 10% DMSO and a thawing rate of 30 °C for 16 seconds or 40 °C for 12 seconds is recommended for P. brevis semen cryopreservation. |
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Semina. Ciências Agrárias (Online) |
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Cryopreservation of Prochilodus brevis semen: freezing media and thawing ratesCriopreservação do sêmen de Prochilodus brevis: meios de congelação e taxas de descongelaçãoCryoprotective agentCurimatã comumSperm extenderSperm freezingSperm qualityThawing temperature.Agente crioprotetorCurimatã comum.Prochilodus brevis is a migratory fish, an important component of its river ecosystem and an appreciated animal in northeastern cuisine. However, human activities have threatened its survival. Thus, researchers have become interested in developing genetic material storage protocols, such as seminal cryopreservation. Therefore, determination of the appropriate freezing media and thawing rate is a fundamental step toward the use of this biotechnology in the production of common curimatã and for reducing risks to the species’ survival. As such, the aim of the current study was to evaluate the effect of different freezing media and thawing rates on the quality of cryopreserved semen from P. brevis. For this study, males received a single dose of pituitary extract of carp 18 hours before semen collection. The semen samples were diluted in 5% glucose + 10% methyl glycol (MG), 5% glucose + 10% dimethyl sulfoxide (DMSO), 0.9% NaCl + 10% methyl glycol, and 0.9% NaCl + 10% dimethyl sulfoxide, loaded into 0.25-mL straws and frozen in liquid nitrogen vapor. The semen from each treatment was thawed at three different thawing rates: 25 °C for 30 sec, 30 °C for 16 sec and 40 °C for 12 sec. Motility, vitality and morphology analyses were performed by computer-assisted sperm analysis (CASA). The characteristics of the fresh sperm mostly resembled those found in the literature. For the parameters analyzed, fresh sperm presented higher sperm quality in comparison to all treatments with cryopreserved sperm (p < 0.05), except for the characteristic of normal morphology, for which the sperm cryopreserved in glucose and MG did not differ statistically from the fresh sperm. For the cryopreserved semen, the greatest results of total motility and curvilinear velocity (VCL) were obtained using glucose and DMSO, regardless of the thawing rate employed. For the straight-line velocity (VSL) and average path velocity (VAP), DMSO showed the best results, regardless of the diluent and thawing rate. With regard to vitality, the highest values were achieved when DMSO and thawing rates of 30 °C for 16 sec or 40 °C for 12 sec were used. In the morphological analysis, the greatest percentage of normal sperm cells was obtained using thawing rates of 25 °C for 30 sec and 40 °C for 12 sec, regardless of the freezing media. Sperm quality was found to suffer interference from the freezing media, as well as from interaction between its components (diluent and cryoprotectant) and the thawing rate used. Under the methodological conditions employed, the use of 5% glucose + 10% DMSO and a thawing rate of 30 °C for 16 seconds or 40 °C for 12 seconds is recommended for P. brevis semen cryopreservation.O Prochilodus brevis é um peixe reofílico, importante componente do ecossistema fluvial e apreciado na culinária nordestina. No entanto, ações antrópicas têm ameaçado sua sobrevivência. Desta forma, surge, nos pesquisadores, o interesse no desenvolvimento de protocolos de conservação do material genético, como a criopreservação seminal. Logo, a determinação do meio de congelação e da taxa de descongelação adequados, são passos fundamentais que possibilitarão a utilização dessa biotecnologia na produção de curimatã comum, reduzindo os riscos à sua sobrevivência. Portanto, o objetivo desse trabalho foi avaliar o efeito de diferentes meios de congelação e taxas de descongelação sobre a qualidade do sêmen criopreservado de P. brevis. Para isso, 18 horas antes da coleta de sêmen, os machos receberam dose única de extrato hipofisário de carpa. Cada animal foi sedado com solução à base de eugenol e o sêmen foi coletado. As amostras foram diluídas em quatro meios de congelação (5% Glicose + Metilglicol 10%; 5% Glicose + DMSO 10%; 0,9% NaCl + Metilglicol 10%; 0,9% NaCl + DMSO 10%) envasadas em palhetas de 0,25 mL e congeladas em vapor de nitrogênio líquido. O sêmen foi descongelado após sete dias em três taxas de descongelação: 25 °C 30 s-1; 30 °C 16 s-1; 40 °C 12 s-1. Foram feitas as análises de motilidade, vitalidade e morfologia com auxílio de sistema automatizado de análise seminal (CASA). As características do sêmen in natura assemelharam-se, em sua maioria, às encontradas na literatura. Para os parâmetros analisados, o sêmen in natura apresentou qualidade seminal superior a todos os tratamentos com o sêmen criopreservado (p < 0,05), exceto para a morfologia normal, no sêmen criopreservado em Glicose e MG, que não diferiu estatisticamente do sêmen in natura (p > 0,05). Para o sêmen criopreservado, os maiores índices foram alcançados quando se utilizou DMSO e as taxas de descongelação de 30 °C 16 s-1 ou 40 °C 12 s-1. Quanto à análise morfológica, a maior porcentagem de espermatozoides normais foi obtida utilizando as taxas de 25 °C 30 s-1 e 40 °C 12 s-1, independente do meio de congelação. A qualidade seminal sofre interferência do meio de congelação, bem como da interação entre seus componentes (diluente e crioprotetor) e da taxa de descongelação empregada. Sob as condições metodológicas empregadas, recomenda-se a criopreservação do sêmen de P. brevis em 5% Glicose + DMSO 10% e descongelação a 30 °C durante 16 segundos ou 40 °C durante 12 segundos.UEL2016-06-22info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionPesquisa Empírica de Campoapplication/pdfhttps://ojs.uel.br/revistas/uel/index.php/semagrarias/article/view/2200310.5433/1679-0359.2016v37n3p1643Semina: Ciências Agrárias; Vol. 37 No. 3 (2016); 1643-1654Semina: Ciências Agrárias; v. 37 n. 3 (2016); 1643-16541679-03591676-546Xreponame:Semina. Ciências Agrárias (Online)instname:Universidade Estadual de Londrina (UEL)instacron:UELenghttps://ojs.uel.br/revistas/uel/index.php/semagrarias/article/view/22003/19048http://creativecommons.org/licenses/by-nc/4.0info:eu-repo/semantics/openAccessNunes, Larissa TeixeiraOliveira, Mayara SetubalLopes, Júlia TrugilioSouza, Maria Eduarda Magalhães dePinheiro, Romulo Roberto RibeiroCampello, Cláudio CabralSalmito-Vanderley, Carminda Sandra Brito2022-12-01T15:36:43Zoai:ojs.pkp.sfu.ca:article/22003Revistahttp://www.uel.br/revistas/uel/index.php/semagrariasPUBhttps://ojs.uel.br/revistas/uel/index.php/semagrarias/oaisemina.agrarias@uel.br1679-03591676-546Xopendoar:2022-12-01T15:36:43Semina. Ciências Agrárias (Online) - Universidade Estadual de Londrina (UEL)false |
dc.title.none.fl_str_mv |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates Criopreservação do sêmen de Prochilodus brevis: meios de congelação e taxas de descongelação |
title |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates |
spellingShingle |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates Nunes, Larissa Teixeira Cryoprotective agent Curimatã comum Sperm extender Sperm freezing Sperm quality Thawing temperature. Agente crioprotetor Curimatã comum. Nunes, Larissa Teixeira Cryoprotective agent Curimatã comum Sperm extender Sperm freezing Sperm quality Thawing temperature. Agente crioprotetor Curimatã comum. |
title_short |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates |
title_full |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates |
title_fullStr |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates |
title_full_unstemmed |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates |
title_sort |
Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates |
author |
Nunes, Larissa Teixeira |
author_facet |
Nunes, Larissa Teixeira Nunes, Larissa Teixeira Oliveira, Mayara Setubal Lopes, Júlia Trugilio Souza, Maria Eduarda Magalhães de Pinheiro, Romulo Roberto Ribeiro Campello, Cláudio Cabral Salmito-Vanderley, Carminda Sandra Brito Oliveira, Mayara Setubal Lopes, Júlia Trugilio Souza, Maria Eduarda Magalhães de Pinheiro, Romulo Roberto Ribeiro Campello, Cláudio Cabral Salmito-Vanderley, Carminda Sandra Brito |
author_role |
author |
author2 |
Oliveira, Mayara Setubal Lopes, Júlia Trugilio Souza, Maria Eduarda Magalhães de Pinheiro, Romulo Roberto Ribeiro Campello, Cláudio Cabral Salmito-Vanderley, Carminda Sandra Brito |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Nunes, Larissa Teixeira Oliveira, Mayara Setubal Lopes, Júlia Trugilio Souza, Maria Eduarda Magalhães de Pinheiro, Romulo Roberto Ribeiro Campello, Cláudio Cabral Salmito-Vanderley, Carminda Sandra Brito |
dc.subject.por.fl_str_mv |
Cryoprotective agent Curimatã comum Sperm extender Sperm freezing Sperm quality Thawing temperature. Agente crioprotetor Curimatã comum. |
topic |
Cryoprotective agent Curimatã comum Sperm extender Sperm freezing Sperm quality Thawing temperature. Agente crioprotetor Curimatã comum. |
description |
Prochilodus brevis is a migratory fish, an important component of its river ecosystem and an appreciated animal in northeastern cuisine. However, human activities have threatened its survival. Thus, researchers have become interested in developing genetic material storage protocols, such as seminal cryopreservation. Therefore, determination of the appropriate freezing media and thawing rate is a fundamental step toward the use of this biotechnology in the production of common curimatã and for reducing risks to the species’ survival. As such, the aim of the current study was to evaluate the effect of different freezing media and thawing rates on the quality of cryopreserved semen from P. brevis. For this study, males received a single dose of pituitary extract of carp 18 hours before semen collection. The semen samples were diluted in 5% glucose + 10% methyl glycol (MG), 5% glucose + 10% dimethyl sulfoxide (DMSO), 0.9% NaCl + 10% methyl glycol, and 0.9% NaCl + 10% dimethyl sulfoxide, loaded into 0.25-mL straws and frozen in liquid nitrogen vapor. The semen from each treatment was thawed at three different thawing rates: 25 °C for 30 sec, 30 °C for 16 sec and 40 °C for 12 sec. Motility, vitality and morphology analyses were performed by computer-assisted sperm analysis (CASA). The characteristics of the fresh sperm mostly resembled those found in the literature. For the parameters analyzed, fresh sperm presented higher sperm quality in comparison to all treatments with cryopreserved sperm (p < 0.05), except for the characteristic of normal morphology, for which the sperm cryopreserved in glucose and MG did not differ statistically from the fresh sperm. For the cryopreserved semen, the greatest results of total motility and curvilinear velocity (VCL) were obtained using glucose and DMSO, regardless of the thawing rate employed. For the straight-line velocity (VSL) and average path velocity (VAP), DMSO showed the best results, regardless of the diluent and thawing rate. With regard to vitality, the highest values were achieved when DMSO and thawing rates of 30 °C for 16 sec or 40 °C for 12 sec were used. In the morphological analysis, the greatest percentage of normal sperm cells was obtained using thawing rates of 25 °C for 30 sec and 40 °C for 12 sec, regardless of the freezing media. Sperm quality was found to suffer interference from the freezing media, as well as from interaction between its components (diluent and cryoprotectant) and the thawing rate used. Under the methodological conditions employed, the use of 5% glucose + 10% DMSO and a thawing rate of 30 °C for 16 seconds or 40 °C for 12 seconds is recommended for P. brevis semen cryopreservation. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-06-22 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Pesquisa Empírica de Campo |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://ojs.uel.br/revistas/uel/index.php/semagrarias/article/view/22003 10.5433/1679-0359.2016v37n3p1643 |
url |
https://ojs.uel.br/revistas/uel/index.php/semagrarias/article/view/22003 |
identifier_str_mv |
10.5433/1679-0359.2016v37n3p1643 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://ojs.uel.br/revistas/uel/index.php/semagrarias/article/view/22003/19048 |
dc.rights.driver.fl_str_mv |
http://creativecommons.org/licenses/by-nc/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
UEL |
publisher.none.fl_str_mv |
UEL |
dc.source.none.fl_str_mv |
Semina: Ciências Agrárias; Vol. 37 No. 3 (2016); 1643-1654 Semina: Ciências Agrárias; v. 37 n. 3 (2016); 1643-1654 1679-0359 1676-546X reponame:Semina. Ciências Agrárias (Online) instname:Universidade Estadual de Londrina (UEL) instacron:UEL |
instname_str |
Universidade Estadual de Londrina (UEL) |
instacron_str |
UEL |
institution |
UEL |
reponame_str |
Semina. Ciências Agrárias (Online) |
collection |
Semina. Ciências Agrárias (Online) |
repository.name.fl_str_mv |
Semina. Ciências Agrárias (Online) - Universidade Estadual de Londrina (UEL) |
repository.mail.fl_str_mv |
semina.agrarias@uel.br |
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1822182751314378752 |
dc.identifier.doi.none.fl_str_mv |
10.5433/1679-0359.2016v37n3p1643 |