ABSTRACT
The action of substances with non-permeable cryoprotectant potential, besides glucose, has not yet been studied for the species Prochilodus brevis. The objective of this work was to evaluate the action of four non-permeable cryoprotectants on this species sperm cryopreservation. Five pools were cryopreserved in a solution of 5% glucose and 10% dimethyl sulfoxide (Me2SO) associated or not (control) with cryoprotectants egg yolk (5, 10 or 12%), soy lecithin (2.5, 7.5 or 10%), sucrose (5, 10 or 20%) and lactose (5, 8 or 15%). After thawing, samples were evaluated for sperm kinetics (total motility, motility duration, velocities, and wobble - WOB), morphology and membrane and DNA integrity. The treatments containing egg yolk improved significantly (P<0.05) results when compared the control for the membrane integrity parameter. When compared to other treatments, egg yolk, at any concentration, presented higher results (P<0.05) for membrane integrity, total motility, curvilinear velocity (VCL) and average path velocity (VAP) parameters. Egg yolk also showed the best results for WOB, but it did not differ from 5% and 8% lactose and 5% and 20% sucrose. Soy lecithin had the lowest percentages of morphologically normal sperm (P<0.05), while the other treatments did not differ from each other. There was no difference regarding DNA integrity data. Thus, 5% egg yolk is indicated as a non-permeable cryoprotectant for P. brevis, in association with 5% glucose and 10% Me2SO.
ABSTRACT
The action of substances with non-permeable cryoprotectant potential, besides glucose, has not yet been studied for the species Prochilodus brevis. The objective of this work was to evaluate the action of four non-permeable cryoprotectants on this species sperm cryopreservation. Five pools were cryopreserved in a solution of 5% glucose and 10% dimethyl sulfoxide (Me2SO) associated or not (control) with cryoprotectants egg yolk (5, 10 or 12%), soy lecithin (2.5, 7.5 or 10%), sucrose (5, 10 or 20%) and lactose (5, 8 or 15%). After thawing, samples were evaluated for sperm kinetics (total motility, motility duration, velocities, and wobble - WOB), morphology and membrane and DNA integrity. The treatments containing egg yolk improved significantly (P<0.05) results when compared the control for the membrane integrity parameter. When compared to other treatments, egg yolk, at any concentration, presented higher results (P<0.05) for membrane integrity, total motility, curvilinear velocity (VCL) and average path velocity (VAP) parameters. Egg yolk also showed the best results for WOB, but it did not differ from 5% and 8% lactose and 5% and 20% sucrose. Soy lecithin had the lowest percentages of morphologically normal sperm (P<0.05), while the other treatments did not differ from each other. There was no difference regarding DNA integrity data. Thus, 5% egg yolk is indicated as a non-permeable cryoprotectant for P. brevis, in association with 5% glucose and 10% Me2SO.(AU)
Subject(s)
Animals , Fishes , Cryopreservation , Characidae , Cryoprotective AgentsABSTRACT
Abstract The action of substances with non-permeable cryoprotectant potential, besides glucose, has not yet been studied for the species Prochilodus brevis. The objective of this work was to evaluate the action of four non-permeable cryoprotectants on this species sperm cryopreservation. Five pools were cryopreserved in a solution of 5% glucose and 10% dimethyl sulfoxide (Me2SO) associated or not (control) with cryoprotectants egg yolk (5, 10 or 12%), soy lecithin (2.5, 7.5 or 10%), sucrose (5, 10 or 20%) and lactose (5, 8 or 15%). After thawing, samples were evaluated for sperm kinetics (total motility, motility duration, velocities, and wobble - WOB), morphology and membrane and DNA integrity. The treatments containing egg yolk improved significantly (P<0.05) results when compared the control for the membrane integrity parameter. When compared to other treatments, egg yolk, at any concentration, presented higher results (P<0.05) for membrane integrity, total motility, curvilinear velocity (VCL) and average path velocity (VAP) parameters. Egg yolk also showed the best results for WOB, but it did not differ from 5% and 8% lactose and 5% and 20% sucrose. Soy lecithin had the lowest percentages of morphologically normal sperm (P<0.05), while the other treatments did not differ from each other. There was no difference regarding DNA integrity data. Thus, 5% egg yolk is indicated as a non-permeable cryoprotectant for P. brevis, in association with 5% glucose and 10% Me2SO.
ABSTRACT
Prochilodus brevis is a fish species with an important ecological and economic role. Therefore, its captive breeding, together with sperm cryopreservation, come as an alternative to improve its availability. To perform sperm cryopreservation with quality results, it is necessary to develop studies on cryomedia composition, such as non-permeable cryoprotectants. The aim of the study was to test the efficacy of two non-permeable cryoprotectants on post-thawed kinetic parameters of Prochilodus brevis sperm. Twenty reproductive mature P. brevis males were selected and induced to sperm with carp pituitary extract. The sperm was collected, and five pools were made. Pools were diluted in 5% glucose and 10% DMSO added or not with three different concentrations of egg yolk (5; 10 or 12%). The samples were placed on French straws, frozen in dry shipper and stored in liquid nitrogen. After thawing, the kinetic analyses of computer assisted sperm analysis (CASA) were performed. There were no significant differences between the total motility of the control (60.48%) and treatments (49.0; 47.8 and 51.68%, respectively). The other kinetic parameters also showed no statistical difference. Thus, there is no need to use egg yolk in the freezing of P. brevis sperm when using a glucose and DMSO extender.
Subject(s)
Male , Animals , Cryopreservation/methods , Cryopreservation/veterinary , Egg Yolk/adverse effects , FishesABSTRACT
Prochilodus brevis is a fish species with an important ecological and economic role. Therefore, its captive breeding, together with sperm cryopreservation, come as an alternative to improve its availability. To perform sperm cryopreservation with quality results, it is necessary to develop studies on cryomedia composition, such as non-permeable cryoprotectants. The aim of the study was to test the efficacy of two non-permeable cryoprotectants on post-thawed kinetic parameters of Prochilodus brevis sperm. Twenty reproductive mature P. brevis males were selected and induced to sperm with carp pituitary extract. The sperm was collected, and five pools were made. Pools were diluted in 5% glucose and 10% DMSO added or not with three different concentrations of egg yolk (5; 10 or 12%). The samples were placed on French straws, frozen in dry shipper and stored in liquid nitrogen. After thawing, the kinetic analyses of computer assisted sperm analysis (CASA) were performed. There were no significant differences between the total motility of the control (60.48%) and treatments (49.0; 47.8 and 51.68%, respectively). The other kinetic parameters also showed no statistical difference. Thus, there is no need to use egg yolk in the freezing of P. brevis sperm when using a glucose and DMSO extender.(AU)
Subject(s)
Animals , Male , Cryopreservation/methods , Cryopreservation/veterinary , Egg Yolk/adverse effects , FishesABSTRACT
A presente pesquisa visou determinar a dose inseminante para a fertilização assistida da carpa comum, e o registro em imagens e o tempo de desenvolvimento de cada fase embrionária desta espécie em latitude equatorial. Verificou-se que a porcentagem de fertilização aumentou de forma linear, atingindo um platô em 45,5% na proporção de 208.295 espermatozoides/oócito. Assim, recomenda-se o uso da dose inseminante de aproximadamente 200.000 espermatozoides/oócito na rotina de fertilização assistida dessa espécie, no nordeste brasileiro. Além disso, o desenvolvimento embrionário da carpa em latitude equatorial segue a cronologia semelhante ao relatado para a espécie em clima temperado.(AU)
The present research aimed at determining insemination dose for artificial fertilization of common carp, and record images and time in the development of each embryonic stage of the species in equatorial latitude. The fertilization rate increased linearly up to the proportion of 208.295 spermatozoa/oocyte, and, from this point, the fertilization rate was maintained at 45.5%. Therefore, we recommend the use of the insemination dose of approximately 200.000 sperm/oocyte in the artificial fertilization routine of common carpin brazilian northeast. Moreover, embryonic development of carp in equatorial latitude follows the chronology similar to that reported for the species in temperate zones.(AU)
Subject(s)
Animals , Carps/embryology , Reproductive Techniques, Assisted/veterinary , Reproductive Control Agents/administration & dosage , Reproduction , Embryonic DevelopmentABSTRACT
A presente pesquisa visou determinar a dose inseminante para a fertilização assistida da carpa comum, e o registro em imagens e o tempo de desenvolvimento de cada fase embrionária desta espécie em latitude equatorial. Verificou-se que a porcentagem de fertilização aumentou de forma linear, atingindo um platô em 45,5% na proporção de 208.295 espermatozoides/oócito. Assim, recomenda-se o uso da dose inseminante de aproximadamente 200.000 espermatozoides/oócito na rotina de fertilização assistida dessa espécie, no nordeste brasileiro. Além disso, o desenvolvimento embrionário da carpa em latitude equatorial segue a cronologia semelhante ao relatado para a espécie em clima temperado.
The present research aimed at determining insemination dose for artificial fertilization of common carp, and record images and time in the development of each embryonic stage of the species in equatorial latitude. The fertilization rate increased linearly up to the proportion of 208.295 spermatozoa/oocyte, and, from this point, the fertilization rate was maintained at 45.5%. Therefore, we recommend the use of the insemination dose of approximately 200.000 sperm/oocyte in the artificial fertilization routine of common carpin brazilian northeast. Moreover, embryonic development of carp in equatorial latitude follows the chronology similar to that reported for the species in temperate zones.
Subject(s)
Animals , Carps/embryology , Reproductive Control Agents/administration & dosage , Reproductive Techniques, Assisted/veterinary , Embryonic Development , ReproductionABSTRACT
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 parameter...(AU)
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...(AU)
Subject(s)
Animals , Characiformes/embryology , Characiformes/physiology , Cryopreservation , Sperm Retrieval , Cryoprotective AgentsABSTRACT
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 parameter...
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...
Subject(s)
Animals , Characiformes/embryology , Characiformes/physiology , Cryopreservation , Cryoprotective Agents , Sperm RetrievalABSTRACT
The objective of the current study was to observe the performance kinetics (motilities and velocities) of the spermatozoa from Prochilodus brevis (curimatã), Colossoma macropomum (tambaqui) and Piaractus brachypomus (pirapitinga) species in different times post-activation. The sperm of P. brevis, C. macropomum and P. brachypomus species were collected after hormonal induction with carp pituitary extract. The samples with not contamination with water, urine or feces had motility subjective, morphology, osmolality and concentration analyzed. The samples selected were analyzed with Sperm Class Analyzer. Spermatozoa motility and velocities were captured at 10, 30, 60 and 120 s postactivation. No significant differences in total motility of P. brevis spermatozoa were observed between 10 s and 30 s post-activation. However, significant reduction was observed in 60 s. This reduction was more accentuated after 120 s. The same pattern of spermatozoa motility decline happened for C. macropomum and P. brachypomus. Velocities also followed the same pattern for the three species. There was significant reduction in velocities after 30 s; this reduction was more significant after 60 s. There was no significance difference between 60 s and 120 s post-activation. Sperm of C. macropomum and P. brachypomus show satisfactory sperm quality up to 60 s after activation. On the other hand, sperm of P. brevis up to 120 s after activation. These findings show that the rate of sperm motility in different times post activation is change for each species tested.(AU)
O objetivo do presente trabalho foi investigar o desempenho cinético (motilidade e velocidades espermáticas) de Prochilodus brevis (curimatã), Colossoma macropomum (tambaqui) e Piaractus brachypomus (pirapitinga) em diferentes tempos pós ativação. O sêmen de P. brevis, C. macropomum e P. brachypomus foi coletado após indução hormonal com extrato hipofisário de carpa. As amostras não contaminadas com sangue, fezes ou urina tiveram motilidade subjetiva, morfologia, osmolaridade e concentração analisadas. As amostras selecionadas foram analisadas com o Sperm Class Analyzer. A motilidade e velocidade dos espermatozoides foram mensuradas a 10, 30, 60 e 120 s pós ativação. Nenhuma diferença significativa na motilidade total do sêmen de P. brevis foi observada entre 10 s e 30 s pós ativação. No entanto, uma redução significativa foi observada aos 60 s. Essa redução foi mais acentuada após 120 s. O mesmo padrão de declínio da motilidade espermática ocorreu para C. macropomum e P. brachypomus. As velocidades espermáticas seguiram o mesmo padrão para as três espécies. Houve redução significativa nas velocidades após 30 s; essa redução foi mais significativa após 60 s. Não houve diferença significativa entre 60 s e 120 s pós ativação. Os espermatozoides de C. macropomum e P. brachypomus apresentam satisfatotia qualidade espermática até 60 s pós ativação. Por outro lado, os espermatozoides de P. brevis apresentam satisfatória qualidade espermática até 120 s pós ativação. Esses achados mostram que a taxa de motilidade espermática em diferentes tempos pós ativação muda para cada espécie testada.(AU)
Subject(s)
Animals , Fishes , Characiformes , Sperm Motility , Semen Analysis/veterinaryABSTRACT
The objective of the current study was to observe the performance kinetics (motilities and velocities) of the spermatozoa from Prochilodus brevis (curimatã), Colossoma macropomum (tambaqui) and Piaractus brachypomus (pirapitinga) species in different times post-activation. The sperm of P. brevis, C. macropomum and P. brachypomus species were collected after hormonal induction with carp pituitary extract. The samples with not contamination with water, urine or feces had motility subjective, morphology, osmolality and concentration analyzed. The samples selected were analyzed with Sperm Class Analyzer. Spermatozoa motility and velocities were captured at 10, 30, 60 and 120 s postactivation. No significant differences in total motility of P. brevis spermatozoa were observed between 10 s and 30 s post-activation. However, significant reduction was observed in 60 s. This reduction was more accentuated after 120 s. The same pattern of spermatozoa motility decline happened for C. macropomum and P. brachypomus. Velocities also followed the same pattern for the three species. There was significant reduction in velocities after 30 s; this reduction was more significant after 60 s. There was no significance difference between 60 s and 120 s post-activation. Sperm of C. macropomum and P. brachypomus show satisfactory sperm quality up to 60 s after activation. On the other hand, sperm of P. brevis up to 120 s after activation. These findings show that the rate of sperm motility in different times post activation is change for each species tested.
O objetivo do presente trabalho foi investigar o desempenho cinético (motilidade e velocidades espermáticas) de Prochilodus brevis (curimatã), Colossoma macropomum (tambaqui) e Piaractus brachypomus (pirapitinga) em diferentes tempos pós ativação. O sêmen de P. brevis, C. macropomum e P. brachypomus foi coletado após indução hormonal com extrato hipofisário de carpa. As amostras não contaminadas com sangue, fezes ou urina tiveram motilidade subjetiva, morfologia, osmolaridade e concentração analisadas. As amostras selecionadas foram analisadas com o Sperm Class Analyzer. A motilidade e velocidade dos espermatozoides foram mensuradas a 10, 30, 60 e 120 s pós ativação. Nenhuma diferença significativa na motilidade total do sêmen de P. brevis foi observada entre 10 s e 30 s pós ativação. No entanto, uma redução significativa foi observada aos 60 s. Essa redução foi mais acentuada após 120 s. O mesmo padrão de declínio da motilidade espermática ocorreu para C. macropomum e P. brachypomus. As velocidades espermáticas seguiram o mesmo padrão para as três espécies. Houve redução significativa nas velocidades após 30 s; essa redução foi mais significativa após 60 s. Não houve diferença significativa entre 60 s e 120 s pós ativação. Os espermatozoides de C. macropomum e P. brachypomus apresentam satisfatotia qualidade espermática até 60 s pós ativação. Por outro lado, os espermatozoides de P. brevis apresentam satisfatória qualidade espermática até 120 s pós ativação. Esses achados mostram que a taxa de motilidade espermática em diferentes tempos pós ativação muda para cada espécie testada.