Seleção de reprodutores caprinos e ovinos para produção de sêmen / Selection of caprine and ovine sires for semen production

A presente revisão propôs analisar quais características são desejáveis para seleção de reprodutores caprinos e ovinos, visando à produção de sêmen. A escolha dos reprodutores deve ser feita de acordo com os padrões exigidos da raça, além da higidez reprodutiva dos animais. Biotecnologias reprodutivas oferecem oportunidades consideráveis para a produção animal, como a inseminação artificial, transferência de embriões e congelamento de sêmen. Análises da produção de espermatozoides são de grande importância, pois está diretamente relacionada com a atividade sexual. A genética do criatório é um fator crítico que influencia o resultado final e a saúde animal. A utilização de marcadores moleculares é uma ferramenta que temos para selecionar características desejáveis em uma prole, através da identificação de biomarcadores. Técnicas de genética molecular potencializam o efeito de biotécnicas reprodutivas e consequentemente a lucratividade da pecuária intensiva.(AU)

The present review proposed to analyze which characteristics are desirable for the selection of goat and sheep breeders, aiming at the production of semen. The choice of breeders must be made in accordance with the standards required for the breed, in addition to the reproductive health of the animals. Reproductive biotechnologies offer considerable opportunities for animal production, such as artificial insemination, embryo transfer and semen freezing. Analyzes of sperm production are of great importance, as it is directly related to sexual activity. Farm genetics is a critical factor that influences the end result and animal health. The use of molecular markers is a tool that we have to select desirable characteristics in an offspring, through the identification of biomarkers. Molecular genetic techniques enhance the effect of reproductive biotechniques and consequently the profitability of intensive livestock farming.(AU)

Goat semen freezing: the two faces of the coin / Congelação de sêmen caprino: as duas faces da moeda

Although the niche market for goat breeding products is on the rise, this activity is still carried out in family and disorganized systems. Thus, technification of this sector is necessary, including the introduction of reproduction biotechnology, such as semen cryopreservation and artificial insemination (AI). Semen freezing enables the breeder to improve the genetic quality of their herds at a lower cost, increasing the male potential without limit of time or space, and facilitating breeding management when in association with AI. As a consequence, the productivity and profitability of this sector is elevated, which could favor the transition from a livestock subsistence system to an industrial one. Nevertheless, the use of frozen semen of goats still presents some limitations, since the extender and procedures used for this biotechnology subject the gametes to numerous structural and functional injuries, both lethal and sublethal. These factors are reflected in the fertility rates, which are reduced after AI with frozen goat semen. In this perspective, the objective of the current review work is to report the positive and negative aspects of semen freezing biotechnology, with emphasis on the goat species.

O nicho consumidor de produtos da caprinocultura se encontra em plena ascensão. Apesar disso, tal atividade pecuária ainda é realizada em sistema familiar e de forma desorganizada. Deste modo, a tecnificação do setor é necessária, inclusive pela introdução de biotécnicas da reprodução, como a criopreservação de sêmen e a inseminação artificial (IA). O uso do sêmen congelado possibilita ao produtor melhorar a qualidade genética de seu rebanho a um menor custo, ampliar o potencial do reprodutor sem limite de tempo ou espaço e facilitar o manejo das criações, quando em associação a IA. Como consequência, são elevadas a produtividade e lucratividade do setor, o que inclusive favorece a transição de um sistema pecuário de subsistência para o industrial. Contudo, a congelação do sêmen caprino e o seu uso ainda apresenta certas limitações, visto que os diluidores e procedimentos empregados para esta biotécnica submetem os gametas à inúmeras injúrias estruturais e funcionais, letais e subletais. Tais fatos se refletem nas taxas de fertilidades, as quais se mostram reduzidas após a IA com sêmen congelado caprino. Diante dessa perspectiva, foi objetivado com este trabalho de revisão expor os pontos positivos e negativos da biotécnica da congelação de sêmen, com ênfase para a espécie caprina.

Goat semen freezing: the two faces of the coin / Congelação de sêmen caprino: as duas faces da moeda

Although the niche market for goat breeding products is on the rise, this activity is still carried out in family and disorganized systems. Thus, technification of this sector is necessary, including the introduction of reproduction biotechnology, such as semen cryopreservation and artificial insemination (AI). Semen freezing enables the breeder to improve the genetic quality of their herds at a lower cost, increasing the male potential without limit of time or space, and facilitating breeding management when in association with AI. As a consequence, the productivity and profitability of this sector is elevated, which could favor the transition from a livestock subsistence system to an industrial one. Nevertheless, the use of frozen semen of goats still presents some limitations, since the extender and procedures used for this biotechnology subject the gametes to numerous structural and functional injuries, both lethal and sublethal. These factors are reflected in the fertility rates, which are reduced after AI with frozen goat semen. In this perspective, the objective of the current review work is to report the positive and negative aspects of semen freezing biotechnology, with emphasis on the goat species.(AU)

O nicho consumidor de produtos da caprinocultura se encontra em plena ascensão. Apesar disso, tal atividade pecuária ainda é realizada em sistema familiar e de forma desorganizada. Deste modo, a tecnificação do setor é necessária, inclusive pela introdução de biotécnicas da reprodução, como a criopreservação de sêmen e a inseminação artificial (IA). O uso do sêmen congelado possibilita ao produtor melhorar a qualidade genética de seu rebanho a um menor custo, ampliar o potencial do reprodutor sem limite de tempo ou espaço e facilitar o manejo das criações, quando em associação a IA. Como consequência, são elevadas a produtividade e lucratividade do setor, o que inclusive favorece a transição de um sistema pecuário de subsistência para o industrial. Contudo, a congelação do sêmen caprino e o seu uso ainda apresenta certas limitações, visto que os diluidores e procedimentos empregados para esta biotécnica submetem os gametas à inúmeras injúrias estruturais e funcionais, letais e subletais. Tais fatos se refletem nas taxas de fertilidades, as quais se mostram reduzidas após a IA com sêmen congelado caprino. Diante dessa perspectiva, foi objetivado com este trabalho de revisão expor os pontos positivos e negativos da biotécnica da congelação de sêmen, com ênfase para a espécie caprina.(AU)

The interference of ozone gas in kinects and mitochondrial potential of equine sperm submitted on cryopreservation

The objective of this study was to evaluate the effects of the addition of different concentrations of ozone to quarter horse semen submitted to cryopreservation. Six ejaculates from four stallions were collected and were divided in four experimental groups: a control group (BotuCRIO® extender) and three other groups with BotuCRIO® ozonized at concentrations of 6, 8 and 12 μg of O3/mL. The semen samples were diluted (200 x 106 spermatozoa/mL), filled in straws and frozen. After thawing (37 ºC, 30s), the samples were evaluated at 0, 30 and 60 minutes of incubation regarding sperm kinetics by a computer-assisted sperm analysis (CASA), and plasma membrane integrity (PMI), acrosome integrity (ACi) and mitochondrial membrane potential (MMP) by fluorescent probes. There was a reduction in the kinetic parameters total motility (TM), progressive motility (PM), curvilinear velocity (VCL), straight line velocity (VSL) and average path velocity (VAP) in all groups during the thermoresistance test (TT), a pattern also found in PMI and MMP analyses (p<0.05). There was no difference (p>0.05) between the control and treatment (6, 8, and 12 μg of O3/mL) groups, in any of the evaluated times for the kinetic parameters TM, linearity (LIN), straightness (STR), wobble index (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF). Regarding the VCL, VSL and VAP parameters, the group treated with 6 μg did not differ from the control or from 8 μg, but was higher than 12 μg at 30 and 60 minutes. ACi and PMI did not differ between groups (p>0.05), but PMI was lower in groups 8 μg and 12 μg compared to the control and 6 μg (p<0.05). It was concluded that the addition of ozone does not present beneficial effects for cryopreservation of equine semen at the concentrations used and decreases important parameters of fertility.(AU)

The interference of ozone gas in kinects and mitochondrial potential of equine sperm submitted on cryopreservation

The objective of this study was to evaluate the effects of the addition of different concentrations of ozone to quarter horse semen submitted to cryopreservation. Six ejaculates from four stallions were collected and were divided in four experimental groups: a control group (BotuCRIO® extender) and three other groups with BotuCRIO® ozonized at concentrations of 6, 8 and 12 μg of O3/mL. The semen samples were diluted (200 x 106 spermatozoa/mL), filled in straws and frozen. After thawing (37 ºC, 30s), the samples were evaluated at 0, 30 and 60 minutes of incubation regarding sperm kinetics by a computer-assisted sperm analysis (CASA), and plasma membrane integrity (PMI), acrosome integrity (ACi) and mitochondrial membrane potential (MMP) by fluorescent probes. There was a reduction in the kinetic parameters total motility (TM), progressive motility (PM), curvilinear velocity (VCL), straight line velocity (VSL) and average path velocity (VAP) in all groups during the thermoresistance test (TT), a pattern also found in PMI and MMP analyses (p0.05) between the control and treatment (6, 8, and 12 μg of O3/mL) groups, in any of the evaluated times for the kinetic parameters TM, linearity (LIN), straightness (STR), wobble index (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF). Regarding the VCL, VSL and VAP parameters, the group treated with 6 μg did not differ from the control or from 8 μg, but was higher than 12 μg at 30 and 60 minutes. ACi and PMI did not differ between groups (p>0.05), but PMI was lower in groups 8 μg and 12 μg compared to the control and 6 μg (p<0.05). It was concluded that the addition of ozone does not present beneficial effects for cryopreservation of equine semen at the concentrations used and decreases important parameters of fertility.

Diluente quimicamente definido e antioxidantes polifenois na criopreservação de sêmen ovino / Chemically defined diluter and polyphenolic antioxidants in the cryopreservation of sheep semen

The objective of this study was to elaborate chemically defined extender based on casein, added with polyphenolic antioxidants, for the freezing of semen from ram breeders. To evaluate Tris-casein (5% glycerol), plus polyphenols, four semen pools from three rams sires were frozen (0µM; 10µM resveratrol; 5µM quercetin; 25µM catechin; 25µM catechin + 10µM resveratrol; 25µM catechin + 5µM quercetin; 10 µM resveratrol + 5µM quercetin; 25µM catechin + 10µM resveratrol + 5µM quercetin). After thawing, the samples were evaluated for sperm kinetics, plasma and acrosomal membrane integrity and mitochondrial membrane potential. There was no significant difference (p≥0.05) between the control group and the treatments for any of the evaluated parameters. However, it can be seen that the Tris-casein extender was efficient in cryopreserving ram semen. It is concluded that the Tris-casein extender can be used for freezing ram semen.

Efeito da adição de nanopartículas de Óxido de Zinco ao diluente de congelação do sêmen ovino / Effect of adding zinc oxide nanoparticles to the freezing extender of ram semen

The aim of this study was to evaluate the effect of adding zinc oxide nanoparticles to the ram semen cryopreservation diluter. Semen pools (n=6), from three Santa Inês breeders, were diluted in Tris-yolk (5% glycerol), supplemented with zinc oxide nanoparticles (0, 25, 75 and 150μg/mL) at a concentration of 200x10 6 sperm/mL. The samples were frozen in an automated system and stored in liquid nitrogen (-196 °C). At the timeof analysis, the semen samples were thawed (37 °C/30s) and evaluated for sperm kinetics, plasma and acrosomal membrane integrity and mitochondrial membrane potential. There was no significant difference (p>0.05) between the experimental groups in the parameters of kinetics, plasma membrane and acrosome integrity. As for the potential of mitochondrial membrane, all treated groups were significantly (p≤0.05) larger than the control. It was concluded that the zinc oxide nanoparticles increase the mitochondrial membrane potential of ram sperm submitted to the freezing/thawing process.

Diluente quimicamente definido e antioxidantes polifenois na criopreservação de sêmen ovino / Chemically defined diluter and polyphenolic antioxidants in the cryopreservation of sheep semen

The objective of this study was to elaborate chemically defined extender based on casein, added with polyphenolic antioxidants, for the freezing of semen from ram breeders. To evaluate Tris-casein (5% glycerol), plus polyphenols, four semen pools from three rams sires were frozen (0µM; 10µM resveratrol; 5µM quercetin; 25µM catechin; 25µM catechin + 10µM resveratrol; 25µM catechin + 5µM quercetin; 10 µM resveratrol + 5µM quercetin; 25µM catechin + 10µM resveratrol + 5µM quercetin). After thawing, the samples were evaluated for sperm kinetics, plasma and acrosomal membrane integrity and mitochondrial membrane potential. There was no significant difference (p≥0.05) between the control group and the treatments for any of the evaluated parameters. However, it can be seen that the Tris-casein extender was efficient in cryopreserving ram semen. It is concluded that the Tris-casein extender can be used for freezing ram semen.(AU)

Efeito da adição de nanopartículas de Óxido de Zinco ao diluente de congelação do sêmen ovino / Effect of adding zinc oxide nanoparticles to the freezing extender of ram semen

The aim of this study was to evaluate the effect of adding zinc oxide nanoparticles to the ram semen cryopreservation diluter. Semen pools (n=6), from three Santa Inês breeders, were diluted in Tris-yolk (5% glycerol), supplemented with zinc oxide nanoparticles (0, 25, 75 and 150μg/mL) at a concentration of 200x10 6 sperm/mL. The samples were frozen in an automated system and stored in liquid nitrogen (-196 °C). At the timeof analysis, the semen samples were thawed (37 °C/30s) and evaluated for sperm kinetics, plasma and acrosomal membrane integrity and mitochondrial membrane potential. There was no significant difference (p>0.05) between the experimental groups in the parameters of kinetics, plasma membrane and acrosome integrity. As for the potential of mitochondrial membrane, all treated groups were significantly (p≤0.05) larger than the control. It was concluded that the zinc oxide nanoparticles increase the mitochondrial membrane potential of ram sperm submitted to the freezing/thawing process.(AU)

Effect of green tea extract (Camellia sinensis) on the spermatic parameters of Wistar rats submitted or not to testicular heat shock

The aim of this study was to evaluate the effect of green tea extract (GTE) on the spermatic parameters of Wistar rats, submitted or not to testicular heat shock (HS). For this, 48 animals were treated according to the experimental groups (G1: not exposed to HS and untreated; G2: exposed to HS and untreated; G3: not exposed to HS and treated with GTE; G4: exposed to HS and treated with GTE). Subgroups of rats were euthanized on days 15, 30, and 60 to recover the spermatozoa. The total motility (TM), vigor, spermatic morphology and concentration, mitochondrial membrane potential, plasma membrane integrity, and acrosome integrity (ACi) were analyzed. The TM was higher in G1 and G3 than in G2 and G4 on day 30, and higher in G4 on day 60. The overall means of TM and vigor were higher in G1 and G3 than in G2 and G4, as well as TM on day 60. For the morphology, G2 and G4 were lower than G1 and G3 on day 15, and G4 was lower than G1 and G3 on day 30. Moreover, in G1 and G3 morphology was higher on days 15 and 30, and in G4 it was lower on day 30, with the overall means being higher in G1 and G3 than in G2 and G4, as well as on days 15 and 60 compared to day 30. The overall mean of ACi, on day 30, was lower than on days 15 and 60 for all the groups. Therefore, HS is shown to be widely deleterious to the gametes, and the daily administration of 100 mg/kg green tea extract does not improve the spermatic parameters of Wistar rats, submitted or not to testicular HS, although it leads to better recovery of spermatic motility and morphology at 60 days.

Effect of green tea extract (Camellia sinensis) on the spermatic parameters of Wistar rats submitted or not to testicular heat shock

The aim of this study was to evaluate the effect of green tea extract (GTE) on the spermatic parameters of Wistar rats, submitted or not to testicular heat shock (HS). For this, 48 animals were treated according to the experimental groups (G1: not exposed to HS and untreated; G2: exposed to HS and untreated; G3: not exposed to HS and treated with GTE; G4: exposed to HS and treated with GTE). Subgroups of rats were euthanized on days 15, 30, and 60 to recover the spermatozoa. The total motility (TM), vigor, spermatic morphology and concentration, mitochondrial membrane potential, plasma membrane integrity, and acrosome integrity (ACi) were analyzed. The TM was higher in G1 and G3 than in G2 and G4 on day 30, and higher in G4 on day 60. The overall means of TM and vigor were higher in G1 and G3 than in G2 and G4, as well as TM on day 60. For the morphology, G2 and G4 were lower than G1 and G3 on day 15, and G4 was lower than G1 and G3 on day 30. Moreover, in G1 and G3 morphology was higher on days 15 and 30, and in G4 it was lower on day 30, with the overall means being higher in G1 and G3 than in G2 and G4, as well as on days 15 and 60 compared to day 30. The overall mean of ACi, on day 30, was lower than on days 15 and 60 for all the groups. Therefore, HS is shown to be widely deleterious to the gametes, and the daily administration of 100 mg/kg green tea extract does not improve the spermatic parameters of Wistar rats, submitted or not to testicular HS, although it leads to better recovery of spermatic motility and morphology at 60 days.(AU)

Correlation between insemination dose and bovine sperm traits on the in vitro embryo production outcomes / Correlação da dose inseminante e características do sêmen bovino no sucesso da produção in vitro de embriões

In the present study we aimed to test the best insemination dose for in vitro embryo production (IVEP) and to correlate sperm traits in bovine. In vitro matured oocytes were inseminated with three different sperm concentrations of the same bull: G1 (1*106), G2 (2*106) and G3 (4*106) sperm/mL. At 18 h post-insemination (hpi), presumptive zygotes [G1 (n=114), G2 (n=139) and G3 (n=136)] were stained to evaluate the pronuclei numbers, or continued to in vitro culture [G1 (n=102), G2 (n=111) and G3 (n=106)]. Sperm kinetics were analyzed using Computer-Assisted Semen Analysis (CASA). Sperm plasma membrane, acrosome integrity and mitochondrial activity were analyzed using fluorescent probes. In vitro fertilization (IVF) and IVEP data were compared using chi-square (P<0.05) and correlated with CASA and fluorescence data using Person Correlation (P<0.05). The IVF efficiency, cleavage and total blastocyst rates did not show any significant difference (P>0.05) among the groups. In G3, the polyspermy rate was the highest (7.4%; P<0.05) without difference (P>0.05) between G1 (0%) and G2 (0%). In G1, the early blastocyst rate was the highest (7.8%; P<0.05), without significant difference (P>0.05) between G2 (1.8%) and G3 (0.9%). The IVF efficiency and total blastocyst rates were positively correlated with curvilinear velocity (VCL) (r≃+1; P<0.05). We concluded that the reduction of insemination dose may negatively affect embryo development and VCL may be used as a parameter to improve the IVEP outcomes.

O objetivo deste estudo foi testar a melhor dose inseminante para a produção de embriões in vitro (IVEP) e sua correlação com as características espermáticas na espécie bovina. Oócitos maturados in vitro foram inseminados com três concentrações diferentes de espermatozoides de único touro: G1 (1*106), G2 (2*106) e G3 (4*106) espermatozoides/mL. Às 18h pós-inseminação (hpi), os presumíveis zigotos [G1 (114), G2 (139) e G3 (136)] foram corados para avaliar o número de pronúcleos, ou continuaram para o cultivo in vitro [G1 (102), G2 (111) e G3 (106)]. Os parâmetros da cinética espermática foram analisados usando o Computer-Assisted Semen Analysis (CASA). A integridade de membrana plasmática espermática, acrossomal e a atividade mitocondrial foram analisadas usando sondas fluorescentes. Os dados da fertilização in vitro (FIV) e IVEP foram comparadas com qui-quadrado (P=0,05) e correlacionados com dados de CASA e Fluorescência usando Correlação de Pearson (r=±1; P<0,05). A eficiência da FIV, taxas de clivagem e blastocisto total não mostraram diferença significativa (P>0,05) entre os grupos. Em G3, a taxa de polispermia foi a maior (7,4%; P<0,05), sem diferença (P>0,05) entre G1 (0%) e G2 (0%). Em G1, a taxa de blastocisto inicial foi a maior (7,8%; P<0,05), sem apresentar diferença significativa (P>0,05) com G2 (1,8%) e G3 (0,9%). A eficiência de FIV e a taxa de blastocisto total foram positivamente correlacionadas com velocidade curvilinear (VCL) (P<0,05). Concluímos que a dose inseminante reduzida pode negativamente afetar o desenvolvimento embrionário e VCL pode ser usada como parâmetro para melhorar os resultados da PEIV.

Correlation between insemination dose and bovine sperm traits on the in vitro embryo production outcomes / Correlação da dose inseminante e características do sêmen bovino no sucesso da produção in vitro de embriões

In the present study we aimed to test the best insemination dose for in vitro embryo production (IVEP) and to correlate sperm traits in bovine. In vitro matured oocytes were inseminated with three different sperm concentrations of the same bull: G1 (1*106 ), G2 (2*106 ) and G3 (4*106 ) sperm/mL. At 18 h post-insemination (hpi), presumptive zygotes [G1 (n=114), G2 (n=139) and G3 (n=136)] were stained to evaluate the pronuclei numbers, or continued to in vitro culture [G1 (n=102), G2 (n=111) and G3 (n=106)]. Sperm kinetics were analyzed using Computer-Assisted Semen Analysis (CASA). Sperm plasma membrane, acrosome integrity and mitochondrial activity were analyzed using fluorescent probes. In vitro fertilization (IVF) and IVEP data were compared using chisquare (P0.05) among the groups. In G3, the polyspermy rate was the highest (7.4%; P0.05) between G1 (0%) and G2 (0%). In G1, the early blastocyst rate was the highest (7.8%; P0.05) between G2 (1.8%) and G3 (0.9%). The IVF efficiency and total blastocyst rates were positively correlated with curvilinear velocity (VCL) (r≃+1; P<0,05). We concluded that the reduction of insemination dose may negatively affect embryo development and VCL may be used as a parameter to improve the IVEP outcomes.(AU)

O objetivo deste estudo foi testar a melhor dose inseminante para a produção de embriões in vitro (IVEP) e sua correlação com as características espermáticas na espécie bovina. Oócitos maturados in vitro foram inseminados com três concentrações diferentes de espermatozoides de único touro: G1 (1*106 ), G2 (2*106 ) e G3 (4*106 ) espermatozoides/mL. Às 18 h pós-inseminação (hpi), os presumíveis zigotos [G1 (114), G2 (139) e G3 (136)] foram corados para avaliar o número de pronúcleos, ou continuaram para o cultivo in vitro [G1 (102), G2 (111) e G3 (106)]. Os parâmetros da cinética espermática foram analisados usando o ComputerAssisted Semen Analysis (CASA). A integridade de membrana plasmática espermática, acrossomal e a atividade mitocondrial foram analisadas usando sondas fluorescentes. Os dados da fertilização in vitro (FIV) e IVEP foram comparadas com qui-quadrado (P=0,05) e correlacionados com dados de CASA e Fluorescência usando Correlação de Pearson (r=±1; P0,05) entre os grupos. Em G3, a taxa de polispermia foi a maior (7,4%; P0,05) entre G1 (0%) e G2 (0%). Em G1, a taxa de blastocisto inicial foi a maior (7,8%; P0,05) com G2 (1,8%) e G3 (0,9%). A eficiência de FIV e a taxa de blastocisto total foram positivamente correlacionadas com velocidade curvilinear (VCL) (P<0,05). Concluímos que a dose inseminante reduzida pode negativamente afetar o desenvolvimento embrionário e VCL pode ser usada como parâmetro para melhorar os resultados da PEIV.(AU)

Correlation between insemination dose and bovine sperm traits on the in vitro embryo production outcomes / Correlação da dose inseminante e características do sêmen bovino no sucesso da produção in vitro de embriões

In the present study we aimed to test the best insemination dose for in vitro embryo production (IVEP) and to correlate sperm traits in bovine. In vitro matured oocytes were inseminated with three different sperm concentrations of the same bull: G1 (1*106 ), G2 (2*106 ) and G3 (4*106 ) sperm/mL. At 18 h post-insemination (hpi), presumptive zygotes [G1 (n=114), G2 (n=139) and G3 (n=136)] were stained to evaluate the pronuclei numbers, or continued to in vitro culture [G1 (n=102), G2 (n=111) and G3 (n=106)]. Sperm kinetics were analyzed using Computer-Assisted Semen Analysis (CASA). Sperm plasma membrane, acrosome integrity and mitochondrial activity were analyzed using fluorescent probes. In vitro fertilization (IVF) and IVEP data were compared using chisquare (P0.05) among the groups. In G3, the polyspermy rate was the highest (7.4%; P0.05) between G1 (0%) and G2 (0%). In G1, the early blastocyst rate was the highest (7.8%; P0.05) between G2 (1.8%) and G3 (0.9%). The IVF efficiency and total blastocyst rates were positively correlated with curvilinear velocity (VCL) (r≃+1; P<0,05). We concluded that the reduction of insemination dose may negatively affect embryo development and VCL may be used as a parameter to improve the IVEP outcomes.

O objetivo deste estudo foi testar a melhor dose inseminante para a produção de embriões in vitro (IVEP) e sua correlação com as características espermáticas na espécie bovina. Oócitos maturados in vitro foram inseminados com três concentrações diferentes de espermatozoides de único touro: G1 (1*106 ), G2 (2*106 ) e G3 (4*106 ) espermatozoides/mL. Às 18 h pós-inseminação (hpi), os presumíveis zigotos [G1 (114), G2 (139) e G3 (136)] foram corados para avaliar o número de pronúcleos, ou continuaram para o cultivo in vitro [G1 (102), G2 (111) e G3 (106)]. Os parâmetros da cinética espermática foram analisados usando o ComputerAssisted Semen Analysis (CASA). A integridade de membrana plasmática espermática, acrossomal e a atividade mitocondrial foram analisadas usando sondas fluorescentes. Os dados da fertilização in vitro (FIV) e IVEP foram comparadas com qui-quadrado (P=0,05) e correlacionados com dados de CASA e Fluorescência usando Correlação de Pearson (r=±1; P0,05) entre os grupos. Em G3, a taxa de polispermia foi a maior (7,4%; P0,05) entre G1 (0%) e G2 (0%). Em G1, a taxa de blastocisto inicial foi a maior (7,8%; P0,05) com G2 (1,8%) e G3 (0,9%). A eficiência de FIV e a taxa de blastocisto total foram positivamente correlacionadas com velocidade curvilinear (VCL) (P<0,05). Concluímos que a dose inseminante reduzida pode negativamente afetar o desenvolvimento embrionário e VCL pode ser usada como parâmetro para melhorar os resultados da PEIV.

Potencial biotecnológico da sexagem espermática na produção animal / Biotechnological potential of sex-sorting sperm in animal production

A predeterminação do sexo da progênie é um desejo entre os criadores, uma vez que permite o direcionamento do sexo dos animais nascidos, segundo a finalidade do sistema de produção. Com a descoberta dos cromossomos sexuais X e Y, estudos para a pré-seleção do sexo seguiram um direcionamento científico, a fim de desenvolver técnicas para a separação das populações de espermatozoides portadores destes cromossomos. Com esta finalidade foi desenvolvido e aprimorado o método de separação em citômetro de fluxo, que é de alta precisão, embora apresente custo elevado e cause injúrias aos espermatozoides. Por conseguinte, protocolos alternativos vêm sendo buscados para viabilizar a sexagem espermática, sem maiores prejuízos estruturais e funcionais aos gametas, no que se destacam os gradientes de densidade de Percoll®. Em decorrência do potencial biotecnológico da sexagem espermática, visando maximizar a produção animal, foi objetivado com esta revisão expor os pontos positivos e negativos das duas principais técnicas usadas para este fim.

The predetermination of progeny sex is a desire among breeders, since it allows the sexing of born animals according the purpose of the production system. With the discovery of the sex chromosomes X and Y, the sex pre-selection studies followed a scientific direction, in order to develop techniques for the separation of sperm populations with these chromosomes. For this purpose it was developed and improved the flow cytometer separation method, which is highly accurate, although presenting a high cost and causing sperm injuries. Therefore, alternative protocols have been sought to enable sperm sexing,without major structural and functional damage to gametes, with emphasis on Percoll® density gradients. Due to the biotechnological potential of sperm sexing, in order to maximize the animal production, it was objectified in this review expose the positive and negative points of the two main techniques used for this purpose.

Potencial biotecnológico da sexagem espermática na produção animal / Biotechnological potential of sex-sorting sperm in animal production

A predeterminação do sexo da progênie é um desejo entre os criadores, uma vez que permite o direcionamento do sexo dos animais nascidos, segundo a finalidade do sistema de produção. Com a descoberta dos cromossomos sexuais X e Y, estudos para a pré-seleção do sexo seguiram um direcionamento científico, a fim de desenvolver técnicas para a separação das populações de espermatozoides portadores destes cromossomos. Com esta finalidade foi desenvolvido e aprimorado o método de separação em citômetro de fluxo, que é de alta precisão, embora apresente custo elevado e cause injúrias aos espermatozoides. Por conseguinte, protocolos alternativos vêm sendo buscados para viabilizar a sexagem espermática, sem maiores prejuízos estruturais e funcionais aos gametas, no que se destacam os gradientes de densidade de Percoll®. Em decorrência do potencial biotecnológico da sexagem espermática, visando maximizar a produção animal, foi objetivado com esta revisão expor os pontos positivos e negativos das duas principais técnicas usadas para este fim.(AU)

The predetermination of progeny sex is a desire among breeders, since it allows the sexing of born animals according the purpose of the production system. With the discovery of the sex chromosomes X and Y, the sex pre-selection studies followed a scientific direction, in order to develop techniques for the separation of sperm populations with these chromosomes. For this purpose it was developed and improved the flow cytometer separation method, which is highly accurate, although presenting a high cost and causing sperm injuries. Therefore, alternative protocols have been sought to enable sperm sexing,without major structural and functional damage to gametes, with emphasis on Percoll® density gradients. Due to the biotechnological potential of sperm sexing, in order to maximize the animal production, it was objectified in this review expose the positive and negative points of the two main techniques used for this purpose.(AU)

Concentration of soybean lecithin affects short-term storage success of goat semen related with seminal plasma removal

The objective of this study was to investigate the need of seminal plasma removal for short-term cooling of buck semen in soybean lecithin (SL) based extender. Each pool was divided equally, and one half was subjected to centrifugation to remove seminal plasma (SP-), while the other half remained with seminal plasma (SP+). Then, both SP+ and SP- samples were diluted in two SL extenders (extender A = 1% SL; extender B = 2% SL), cooled to 5ºC and stored for 48 hours. The sperm kinetics, evaluated by CASA, and plasma membrane integrity (PMI), acrosomal integrity (ACI) and high mitochondrial membrane potential (HMMP), evaluated by epifluorescence microscopy, were determined within five minutes after reaching 5°C (T0), as well as after 24 (T24) and 48 (T48) hours of storage. Interactions (seminal plasma vs. extender vs. time;) were observed for all variables assessed. Total and progressive motility and other variables of sperm kinetics decreased after 24 hours of cooling in the SP+ group, and after 48 hours of storage, these same variables were lower in SP+/B compared to SP-/B groups. Furthermore, SP+ reduced PMI (extender B, T48), HMMP (A and B extenders, T48) and ACI (extender A, T0) compared to SP- samples. The interactions between seminal plasma and soybean lecithin phospholipids seemed to occur in a time-dependent manner. It was concluded that the removal of seminal plasma improves the quality of goat semen that was cooled in a soybean lecithin-based extender, especially when using 2% soybean lecithin.(AU)

Concentration of soybean lecithin affects short-term storage success of goat semen related with seminal plasma removal

The objective of this study was to investigate the need of seminal plasma removal for short-term cooling of buck semen in soybean lecithin (SL) based extender. Each pool was divided equally, and one half was subjected to centrifugation to remove seminal plasma (SP-), while the other half remained with seminal plasma (SP+). Then, both SP+ and SP- samples were diluted in two SL extenders (extender A = 1% SL; extender B = 2% SL), cooled to 5ºC and stored for 48 hours. The sperm kinetics, evaluated by CASA, and plasma membrane integrity (PMI), acrosomal integrity (ACI) and high mitochondrial membrane potential (HMMP), evaluated by epifluorescence microscopy, were determined within five minutes after reaching 5°C (T0), as well as after 24 (T24) and 48 (T48) hours of storage. Interactions (seminal plasma vs. extender vs. time;) were observed for all variables assessed. Total and progressive motility and other variables of sperm kinetics decreased after 24 hours of cooling in the SP+ group, and after 48 hours of storage, these same variables were lower in SP+/B compared to SP-/B groups. Furthermore, SP+ reduced PMI (extender B, T48), HMMP (A and B extenders, T48) and ACI (extender A, T0) compared to SP- samples. The interactions between seminal plasma and soybean lecithin phospholipids seemed to occur in a time-dependent manner. It was concluded that the removal of seminal plasma improves the quality of goat semen that was cooled in a soybean lecithin-based extender, especially when using 2% soybean lecithin.

Testicular and kinetics parameters of spermatozoa from ovines fed with growing levels of hay and alfafa (Medicago sativa) / Parâmetros testiculares e de cinética de espermatozoides de ovinos alimentados com níveis crescentes de feno de alfafa (Medicago sativa)

The objective was to evaluate the inclusion of alfalfa hay in diets for growing sheep on testicular parameters and sperm kinetics sheep. Forty non-castrated male sheep were used, with body weight around 26 kg, ± 1.85 and eight months of age, approximately, evaluated for 100 days (it being 44 days for adaptation at management more diets and 56 for fattening) in feedlot. Treatments were four levels of alfalfa hay 0%, 20%, 40% and 60 %, so that was maintained proportion 60% of roughage for all rations. The type of diet offered to sheep did not affect testicular weight and serum testosterone concentration. The kinetics of sperm ejaculated and epididymal semen recovered also not influenced by the experimental diets. The inclusion of alfalfa hay in sheep diet did not change testicular parameters as well as the kinetics of ejaculated sperm and recovered from epididymal sperm tail may be recommended for nutritional management of breeding.(AU)

Objetivou-se avaliar a inclusão de feno de alfafa em rações para ovinos em crescimento sobre parâmetros testiculares e de cinética espermática de ovinos. Utilizaram-se 40 ovinos em crescimento, com peso corporal médio inicial de 26 ± 1,85 kg e oito meses de idade, aproximadamente, avaliados durante 100 dias (sendo 44 dias para adaptação ao manejo e 56 dias para a terminação) em confinamento. Os tratamentos foram compostos por quatro níveis de feno alfafa: 0%, 20%, 40% e 60%, de modo que foi mantida proporção de 60% de volumoso para todas as rações. O tipo de dieta oferecida aos ovinos não influenciou o peso testicular, bem como a concentração sérica de testosterona.  A cinética espermática do ejaculado e do sêmen recuperado do epidídimo também não sofreu influência das dietas experimentais. A inclusão de feno de alfafa na dieta de ovinos não altera parâmetros testiculares, bem como a cinética espermática do ejaculado e também dos espermatozoides recuperados da cauda do epidídimo, podendo ser recomendado em manejo nutricional de reprodutores.(AU)

Testicular and kinetics parameters of spermatozoa from ovines fed with growing levels of hay and alfafa (Medicago sativa) / Parâmetros testiculares e de cinética de espermatozoides de ovinos alimentados com níveis crescentes de feno de alfafa (Medicago sativa)

The objective was to evaluate the inclusion of alfalfa hay in diets for growing sheep on testicular parameters and sperm kinetics sheep. Forty non-castrated male sheep were used, with body weight around 26 kg, ± 1.85 and eight months of age, approximately, evaluated for 100 days (it being 44 days for adaptation at management more diets and 56 for fattening) in feedlot. Treatments were four levels of alfalfa hay 0%, 20%, 40% and 60 %, so that was maintained proportion 60% of roughage for all rations. The type of diet offered to sheep did not affect testicular weight and serum testosterone concentration. The kinetics of sperm ejaculated and epididymal semen recovered also not influenced by the experimental diets. The inclusion of alfalfa hay in sheep diet did not change testicular parameters as well as the kinetics of ejaculated sperm and recovered from epididymal sperm tail may be recommended for nutritional management of breeding.

Objetivou-se avaliar a inclusão de feno de alfafa em rações para ovinos em crescimento sobre parâmetros testiculares e de cinética espermática de ovinos. Utilizaram-se 40 ovinos em crescimento, com peso corporal médio inicial de 26 ± 1,85 kg e oito meses de idade, aproximadamente, avaliados durante 100 dias (sendo 44 dias para adaptação ao manejo e 56 dias para a terminação) em confinamento. Os tratamentos foram compostos por quatro níveis de feno alfafa: 0%, 20%, 40% e 60%, de modo que foi mantida proporção de 60% de volumoso para todas as rações. O tipo de dieta oferecida aos ovinos não influenciou o peso testicular, bem como a concentração sérica de testosterona.  A cinética espermática do ejaculado e do sêmen recuperado do epidídimo também não sofreu influência das dietas experimentais. A inclusão de feno de alfafa na dieta de ovinos não altera parâmetros testiculares, bem como a cinética espermática do ejaculado e também dos espermatozoides recuperados da cauda do epidídimo, podendo ser recomendado em manejo nutricional de reprodutores.