O que análises avançadas do sêmen podem contribuir de fato para a os resultados da IA em suínos? / What advanced semen analyzes can contribute to AI results in pigs?

O controle da qualidade seminal produzido por centrais e unidades de disseminação genética permite a maximização da produção de doses inseminantes por ejaculado, bem como organiza o acompanhamento da produtividade de cada macho. Este acompanhamento é crítico tanto quando o reprodutor jovem é introduzido na rotina de coleta de sêmen quanto na determinação do momento do descarte e reposição. Essencialmente, a avaliação do sêmen em suínos é a mesma há décadas, porém novas metodologias de análise individual da célula como a citometria de fluxo e de constituintes de tecidos e fluídos como a proteômica e metabolômica já estão trazendo avanços na andrologia animal. Na presente revisão, são abordados os aspectos técnicos, vantagens e limitações destas três análises avançadas aplicadas ao sêmen suíno, discutindo a sua implementação e impactos no manejo reprodutivo da espécie.(AU)

The quality control of the semen doses produced by artificial insemination centers and allows the maximization of the production of inseminating doses per ejaculate, as well as organizes the monitoring of the productivity of each male. This follow-up is critical when the young male is initiated in routine semen collection and in determining the moment of male replacement. Essentially, semen evaluation in pigs has been the same for decades, but new methodologies for individual cell analysis such as flow cytometry and tissue and fluid constituents such as proteomics and metabolomics are already bringing advances in animal andrology. In this review, the technical aspects, advantages, and limitations of these three advanced analyzes applied to swine semen are addressed, discussing their implementation and impacts on the reproductive management of the species.(AU)

Reproductive parameters of lambs immunocastrated with anti-GnRH vaccine

Abstract The objective of this study was to evaluate the testicular biometric, seminal, and plasma testosterone levels in lambs subjected to an anti-GnRH vaccine as a method of castration. Thirty entire, crossbred Santa Inês male lambs were randomly distributed into three treatment (T): T1 was the control group, with the administration of 1 mL of saline solution subcutaneously (SC); 1.0 and 0.5 mL of an anti-GnRH vaccine were administered SC in T2 and T3, respectively. Testicular biometric variables, physical and morphological variables of semen, and plasma testosterone concentrations were evaluated. At D60, there was a reduction in testicular length, width, thickness, and scrotal circumference of the immunocastrated animals regardless of the vaccine dose used (P < 0.05). A reduction in semen physical variables at both dosages (P < 0.05) was observed, with azoospermia, in 80% and 70% of animals in the T2 and T3 groups, respectively. At D60, the immunocastrated animals also showed an increase in spermatozoa defects (P < 0.05), whereas plasma testosterone concentration decreased (P < 0.05). Immunocastration of lambs using the Bopriva vaccine at doses of 1.0 and 0.5 mL is efficient in inducing azoospermia in up to 80% of animals, although two doses in a 30-day interval are necessary for it to be an effective and safe method. Efficacy was demonstrated through a reduction in serum testosterone levels, testicular biometry, and seminal fluid analysis. Considering the efficacy of both doses in this study, we recommend using the lower dose (0.5 mL), which will allow for a 50% reduction in vaccine costs.

Reproductive parameters of lambs immunocastrated with anti-GnRH vaccine

The objective of this study was to evaluate the testicular biometric, seminal, and plasma testosterone levels in lambs subjected to an anti-GnRH vaccine as a method of castration. Thirty entire, crossbred Santa Inês male lambs were randomly distributed into three treatment (T): T1 was the control group, with the administration of 1 mL of saline solution subcutaneously (SC); 1.0 and 0.5 mL of an anti-GnRH vaccine were administered SC in T2 and T3, respectively. Testicular biometric variables, physical and morphological variables of semen, and plasma testosterone concentrations were evaluated. At D60, there was a reduction in testicular length, width, thickness, and scrotal circumference of the immunocastrated animals regardless of the vaccine dose used (P < 0.05). A reduction in semen physical variables at both dosages (P < 0.05) was observed, with azoospermia, in 80% and 70% of animals in the T2 and T3 groups, respectively. At D60, the immunocastrated animals also showed an increase in spermatozoa defects (P < 0.05), whereas plasma testosterone concentration decreased (P < 0.05). Immunocastration of lambs using the Bopriva vaccine at doses of 1.0 and 0.5 mL is efficient in inducing azoospermia in up to 80% of animals, although two doses in a 30-day interval are necessary for it to be an effective and safe method. Efficacy was demonstrated through a reduction in serum testosterone levels, testicular biometry, and seminal fluid analysis. Considering the efficacy of both doses in this study, we recommend using the lower dose (0.5 mL), which will allow for a 50% reduction in vaccine costs.(AU)

Purificação de fração enriquecida de binder of sperm protein 1 (BSP 1) do fluido seminal bovino através da técnica de precipitação por sulfato de amônio / Purification of enriched fraction of binder of sperm protein 1 (BSP1) from bovine seminal fluid by ammonium sulphate precipitation technique

Objetivou-se no presente estudo realizar purificação parcial da proteína binder of sperm protein1 (BSP1), proteína de 15 kDa presente nos fluidos das glândulas vesiculares de bovinos, por meio deprecipitação com sulfato de amônio. O fluido seminal foi extraído das glândulas vesiculares obtidas dedez touros bos indicus em abatedouro comercial de Fortaleza, Ceará. As glândulas foram transportadas eprocessadas no Laboratório de Fisiologia Animal da Universidade Federal do Ceará. Foi utilizado umcoquetel inibidor de proteases, com posterior centrifugação. Um pool de todas as amostras foiarmazenado a -20 ºC e uma alíquota foi reservada para determinar a concentração de proteína. Para aprecipitação de proteínas, foram adicionadas diferentes concentrações de sulfato de amônio. Foramrealizados SDS-PAGE e Western blot com anticorpo específico. Obtivemos a presença de BSP1. Foiobservada presença da proteína em todas as frações, porém, a fração 40-60% foi a que apresentou a maiorconcentração de BSP1, em média 98%. A técnica utilizada foi altamente eficaz e a purificação dessasproteínas possibilita uma melhoria na qualidade seminal in vitro e pode ser utilizada para obtenção deamostras altamente enriquecidas de BSPs, além de avaliar o potencial de fertilidade de reprodutores.(AU)

The objective of the present study was to partially purify binder of sperm protein 1 (BSP1)protein, a 15 kDa protein present in bovine vesicular gland fluids, by precipitation with ammoniumsulfate. Seminal fluid was extracted from the vesicular glands obtained from ten bos indicus bulls in acommercial slaughterhouse in Fortaleza, Ceará. The glands were transported and processed at theAnimal Physiology Laboratory of the Federal University of Ceará. A protease inhibitor cocktail wasused, with subsequent centrifugation. A pool of all samples was stored at -20 ° C and an aliquot was setaside to determine protein concentration. For protein precipitation, different concentrations ofammonium sulfate were added. SDS-PAGE and Western blot were performed with specific antibody. Weobtained the presence of BSP1. Protein presence was observed in all fractions, however, the 40-60%fraction presented the highest BSP1 concentration, on average 98%. The technique used was highlyeffective and the purification of these proteins allows an improvement in seminal quality in vitro and canbe used to obtain highly enriched samples of BSPs, besides evaluating the fertility potential of breeders.(AU)

Purificação de fração enriquecida de binder of sperm protein 1 (BSP 1) do fluido seminal bovino através da técnica de precipitação por sulfato de amônio / Purification of enriched fraction of binder of sperm protein 1 (BSP1) from bovine seminal fluid by ammonium sulphate precipitation technique

Objetivou-se no presente estudo realizar purificação parcial da proteína binder of sperm protein1 (BSP1), proteína de 15 kDa presente nos fluidos das glândulas vesiculares de bovinos, por meio deprecipitação com sulfato de amônio. O fluido seminal foi extraído das glândulas vesiculares obtidas dedez touros bos indicus em abatedouro comercial de Fortaleza, Ceará. As glândulas foram transportadas eprocessadas no Laboratório de Fisiologia Animal da Universidade Federal do Ceará. Foi utilizado umcoquetel inibidor de proteases, com posterior centrifugação. Um pool de todas as amostras foiarmazenado a -20 ºC e uma alíquota foi reservada para determinar a concentração de proteína. Para aprecipitação de proteínas, foram adicionadas diferentes concentrações de sulfato de amônio. Foramrealizados SDS-PAGE e Western blot com anticorpo específico. Obtivemos a presença de BSP1. Foiobservada presença da proteína em todas as frações, porém, a fração 40-60% foi a que apresentou a maiorconcentração de BSP1, em média 98%. A técnica utilizada foi altamente eficaz e a purificação dessasproteínas possibilita uma melhoria na qualidade seminal in vitro e pode ser utilizada para obtenção deamostras altamente enriquecidas de BSPs, além de avaliar o potencial de fertilidade de reprodutores.

The objective of the present study was to partially purify binder of sperm protein 1 (BSP1)protein, a 15 kDa protein present in bovine vesicular gland fluids, by precipitation with ammoniumsulfate. Seminal fluid was extracted from the vesicular glands obtained from ten bos indicus bulls in acommercial slaughterhouse in Fortaleza, Ceará. The glands were transported and processed at theAnimal Physiology Laboratory of the Federal University of Ceará. A protease inhibitor cocktail wasused, with subsequent centrifugation. A pool of all samples was stored at -20 ° C and an aliquot was setaside to determine protein concentration. For protein precipitation, different concentrations ofammonium sulfate were added. SDS-PAGE and Western blot were performed with specific antibody. Weobtained the presence of BSP1. Protein presence was observed in all fractions, however, the 40-60%fraction presented the highest BSP1 concentration, on average 98%. The technique used was highlyeffective and the purification of these proteins allows an improvement in seminal quality in vitro and canbe used to obtain highly enriched samples of BSPs, besides evaluating the fertility potential of breeders.

Intervalos de coletas de sêmen em reprodutores suínos no início da vida reprodutiva: avaliação dos parâmetros seminais / Intervals of semen collection from boars during early reproductive life: evaluation of seminal fluid parameters

Background: In swine production, good reproduction rates can be achieved through genetic selection and reproductivebiotechnologies. One of these biotechnologies is artificial insemination, which contributes to disseminate genes and optimizebreeding boars, thus improving the quality of insemination doses. This study focused on evaluating the intervals betweensemen collection from boars at the beginning of their reproductive maturity vis-à-vis the viability of insemination doses.Materials, Methods & Results: Twenty 9-month-old boars of the genetic lineage AGPIC 337 (Agroceres PIC) were used inthis study. The experimental design used here was completely randomized, and the randomly selected males were dividedinto four treatment groups, which were named according to the interval between semen sample collections: T1: 2 days; T2:3 days; T3: 4 days and T4: 7 days. Each treatment comprised 5 animals, and at the end of the 90 days of this study, a totalof 150 ejaculates were obtained in T1, 110 in T2, 90 in T3 and 60 in T4. The values of total motility, volume and spermconcentration of the ejaculates were evaluated, as was oxidative stress by means of thiobarbituric acid reactive substances(TBARS), the latter after 0, 72, 120 and 168 h of the study. Membrane integrity was evaluated at 0, 72, 120 and 168 h usingthe eosin-nigrosin staining procedure. Sperm heat resistance was tested after 120 h, and sperm morphology after 72, 120 and168 h. Sperm concentrations differed, with T3 showing 27.04% and 29.65% higher concentrations (P < 0.05) than groupsT2 and T1, respectively. Total motility in group T4 was 0.56%, 1.98% and 3.28% higher (P < 0.05) than in T3, T2 and T1,respectively, indicating that the 7-day interval produced the best result. The heat...

Intervalos de coletas de sêmen em reprodutores suínos no início da vida reprodutiva: avaliação dos parâmetros seminais / Intervals of semen collection from boars during early reproductive life: evaluation of seminal fluid parameters

Background: In swine production, good reproduction rates can be achieved through genetic selection and reproductivebiotechnologies. One of these biotechnologies is artificial insemination, which contributes to disseminate genes and optimizebreeding boars, thus improving the quality of insemination doses. This study focused on evaluating the intervals betweensemen collection from boars at the beginning of their reproductive maturity vis-à-vis the viability of insemination doses.Materials, Methods & Results: Twenty 9-month-old boars of the genetic lineage AGPIC 337 (Agroceres PIC) were used inthis study. The experimental design used here was completely randomized, and the randomly selected males were dividedinto four treatment groups, which were named according to the interval between semen sample collections: T1: 2 days; T2:3 days; T3: 4 days and T4: 7 days. Each treatment comprised 5 animals, and at the end of the 90 days of this study, a totalof 150 ejaculates were obtained in T1, 110 in T2, 90 in T3 and 60 in T4. The values of total motility, volume and spermconcentration of the ejaculates were evaluated, as was oxidative stress by means of thiobarbituric acid reactive substances(TBARS), the latter after 0, 72, 120 and 168 h of the study. Membrane integrity was evaluated at 0, 72, 120 and 168 h usingthe eosin-nigrosin staining procedure. Sperm heat resistance was tested after 120 h, and sperm morphology after 72, 120 and168 h. Sperm concentrations differed, with T3 showing 27.04% and 29.65% higher concentrations (P < 0.05) than groupsT2 and T1, respectively. Total motility in group T4 was 0.56%, 1.98% and 3.28% higher (P < 0.05) than in T3, T2 and T1,respectively, indicating that the 7-day interval produced the best result. The heat...(AU)

Use of a colloid to optimize centrifugation in the selection of bovine sperm for IVF / Uso de um coloide para otimizar a centrifugação na seleção de espermatozoides bovinos para FIV

This study aimed to evaluate the effect of the force and duration of centrifugation and the impact of cushioned centrifugation on sperm selection by Percoll gradient, on sperm quality and development kinetics of in vitro produced bovine embryos. Two experiments were performed. In Experiment I, a pool of semen was selected by Percoll gradients and the pellet was divided into four groups and distributed in a 2 × 2 factorial, with two forces (2200 × g or 9000 × g) and two durations (1 min or 3 min) of centrifugation. In Experiment II, semen was divided into two groups and selected by Percoll gradient with Cushion Fluid (CF) or without CF (Control) in the second centrifugation. The morphofunctionality, biochemical characteristics and fertilizing capacity of the selected sperms were evaluated. In addition, the development of the resulting bovine embryos was monitored for 48 h post-insemination. Duncan and Chi-square tests (P < 0.05) were used to compare the means. In Experiment I, there was a significant increase in sperm vigor (P < 0.05) after sperm selection in all treatments. The force and duration of centrifugation did not have any effect on sperm motility, vigor, and recovery rate among the different treatments (P > 0.05). In Experiment II, the recovery rate and reactive oxygen species (ROS) production in semen were similar among treatments (P > 0.05) although a higher ROS production was observed in the CF fertilization medium. Total fertilization rate was superior in the CF group (65.4 ± 5.3%) compared to that in Control (39.6 ± 4.9%). However, the normal fertilization and cleavage rate did not differ between the Control (94 ± 6.3% and 58.3 ± 8.3%) and CF (89 ± 7.1% and 75.0 ± 7.3%) groups. The reduction in the force and duration of centrifugation did not decrease the sperm recovery during selection by the Percoll gradient and the use of CF in the second centrifugation did not affect the normal fertilization and development of bovine IVF embryos up to 48 h.(AU)

Esse estudo objetivou avaliar o efeito da força e duração da centrifugação, e o impacto da centrifugação amortecida na seleção espermática por gradientes de Percoll, na qualidade espermática e cinética do desenvolvimento de embriões bovinos produzidos in vitro. Dois experimentos foram realizados. No experimento I, um pool de sêmen foi selecionado por gradientes de Percoll e o pellet dividido em quatro grupos e distribuído em um fatorial 2 x 2, com duas forças (2200 e 9000 X g) e dois tempos (1 e 3 min) de centrifugação. No Experimento II, o sêmen foi dividido em dois grupos e selecionado com (CF) ou sem CushionFluid (Controle) na segunda centrifugação. A morfofuncionalidade, características bioquímicas e capacidade fecundante dos espermatozoides selecionados foram avaliadas. Além disso, o desenvolvimento dos embriões bovinos resultantes foi monitorado por 48 horas pós-inseminação. Os testes Duncan e Qui-quadrado (P<0,05) foram usados para comparar as médias. No Experimento I, houve um aumento significativo no vigor após a seleção espermática para todos os tratamentos. A força e a duração da centrifugação não tiveram nenhum efeito na motilidade, vigor e recuperação espermática entre os diferentes tratamentos (P > 0.05). No Experimento II, a taxa de recuperação e a produção de espécies reativas de oxigênio (EROs) no sêmen foram similares entre os tratamentos (P>0,05), embora a maior produção de EROs foi observada no meio de fecundação do grupo CF. A taxa de fecundação total, foi superior no grupo CF (65.4±5.3%) comparada com o Controle (39.6±4.9%). Entretanto a fecundação normal e a taxa de clivagem não diferiram entre os grupos Controle (94±6.3% e 58.3±8.3%) e CF (89±7.1% and 75.0±7.3%). A redução na força e duração da centrifugação não diminuiu a recuperação espermática durante a seleção por gradientes de Percoll e o uso de CF na segunda centrifugação não influenciou a fecundação normal e o desenvolvimento de embriões bovinos PIV até 48 horas.(AU)

Aspectos Da Técnica De Eletroforese Do Plasma Seminal Em Bovinos / Aspects Of The Technique Of Seminal Plasma Electrophoresis In Bovine / Aspectos De La Técnica De Eletroforesis Del Plasma Seminal En Bovinos

O plasma seminal contém fatores protéicos específicos que provocam importantes efeitos tanto na capacidade de fertilização do espermatozoide quanto na fisiologia reprodutiva da fêmea. O isolamento e caracterização dos componentes do plasma seminal possibilitam a avaliação de sua influência sobre as células espermáticas, o aprimoramento das técnicas de armazenamento e a identificação precoce de animais férteis em relação àqueles subférteis. Por meio das técnicas de biologia molecular, muitas proteínas foram identificadas e caracterizadas e o estudo dessas proteínas poderá contribuir para uma melhor compreensão dos processos fisiológicos ligados à fertilidade. Estes auxiliam o diagnóstico de algumas patologias ou diferenciação de animais quanto ao grau de sua fertilidade frente a alterações de clima e manejo, associadas a patologias do trato reprodutivo masculino ou do espermatozoide. O objetivo da presente revisão de literatura é de abordar alguns aspectos do uso da eletroforese do plasma seminal em bovinos para fins de auxílio na seleção de animais para a reprodução.(AU)

Seminal plasma contains specific protein factors that have important effects on both sperm fertilization capacity and female reproductive physiology. The isolation and characterization of the components of the seminal plasma allows the evaluation of their influence on sperm cells, the improvement of storage techniques and the early identification of fertile animals in relation to the subfertile. Through molecular biology techniques, many proteins have been identified and characterized, and the study of these proteins may contribute to a better understanding of physiological processes related to fertility. These help the diagnosis of some pathologies or differentiation of animals as to the degree of their fertility in the face of changes in climate and management, associated with pathologies of the male reproductive tract or spermatozoa. The objective of the present literature review is to address some aspects of the use of seminal plasma electrophoresis in cattle for the purpose of aiding the selection of animals for reproduction.(AU)

El plasma seminal contiene factores proteicos específicos que provocan importantes efectos tanto en la capacidad de fertilización del espermatozoide y en la fisiología reproductiva de la hembra. El aislamiento y caracterización de los componentes del plasma seminal posibilitan la evaluación de su influencia sobre las células espermáticas, el perfeccionamiento de las técnicas de almacenamiento y la identificación precoz de animales fértiles en relación a aquellos subfértiles. A través de las técnicas de biología molecular, muchas proteínas han sido identificadas y caracterizadas y el estudio de estas proteínas puede contribuir a una mejor comprensión de los procesos fisiológicos relacionados con la fertilidad. Estos auxilian el diagnóstico de algunas patologías o diferenciación de animales en cuanto al grado de su fertilidad frente a alteraciones de clima y manejo, asociadas a patologías del tracto reproductivo masculino o del espermatozoide. El objetivo de la presente revisión de literatura es abordar algunos aspectos del uso de la electroforesis del plasma seminal en bovinos para fines de auxilio en la selección de animales para la reproducción.(AU)

Aspectos Da Técnica De Eletroforese Do Plasma Seminal Em Bovinos / Aspects Of The Technique Of Seminal Plasma Electrophoresis In Bovine / Aspectos De La Técnica De Eletroforesis Del Plasma Seminal En Bovinos

O plasma seminal contém fatores protéicos específicos que provocam importantes efeitos tanto na capacidade de fertilização do espermatozoide quanto na fisiologia reprodutiva da fêmea. O isolamento e caracterização dos componentes do plasma seminal possibilitam a avaliação de sua influência sobre as células espermáticas, o aprimoramento das técnicas de armazenamento e a identificação precoce de animais férteis em relação àqueles subférteis. Por meio das técnicas de biologia molecular, muitas proteínas foram identificadas e caracterizadas e o estudo dessas proteínas poderá contribuir para uma melhor compreensão dos processos fisiológicos ligados à fertilidade. Estes auxiliam o diagnóstico de algumas patologias ou diferenciação de animais quanto ao grau de sua fertilidade frente a alterações de clima e manejo, associadas a patologias do trato reprodutivo masculino ou do espermatozoide. O objetivo da presente revisão de literatura é de abordar alguns aspectos do uso da eletroforese do plasma seminal em bovinos para fins de auxílio na seleção de animais para a reprodução.

Seminal plasma contains specific protein factors that have important effects on both sperm fertilization capacity and female reproductive physiology. The isolation and characterization of the components of the seminal plasma allows the evaluation of their influence on sperm cells, the improvement of storage techniques and the early identification of fertile animals in relation to the subfertile. Through molecular biology techniques, many proteins have been identified and characterized, and the study of these proteins may contribute to a better understanding of physiological processes related to fertility. These help the diagnosis of some pathologies or differentiation of animals as to the degree of their fertility in the face of changes in climate and management, associated with pathologies of the male reproductive tract or spermatozoa. The objective of the present literature review is to address some aspects of the use of seminal plasma electrophoresis in cattle for the purpose of aiding the selection of animals for reproduction.

El plasma seminal contiene factores proteicos específicos que provocan importantes efectos tanto en la capacidad de fertilización del espermatozoide y en la fisiología reproductiva de la hembra. El aislamiento y caracterización de los componentes del plasma seminal posibilitan la evaluación de su influencia sobre las células espermáticas, el perfeccionamiento de las técnicas de almacenamiento y la identificación precoz de animales fértiles en relación a aquellos subfértiles. A través de las técnicas de biología molecular, muchas proteínas han sido identificadas y caracterizadas y el estudio de estas proteínas puede contribuir a una mejor comprensión de los procesos fisiológicos relacionados con la fertilidad. Estos auxilian el diagnóstico de algunas patologías o diferenciación de animales en cuanto al grado de su fertilidad frente a alteraciones de clima y manejo, asociadas a patologías del tracto reproductivo masculino o del espermatozoide. El objetivo de la presente revisión de literatura es abordar algunos aspectos del uso de la electroforesis del plasma seminal en bovinos para fines de auxilio en la selección de animales para la reproducción.

Exame andrológico de bubalinos / Andrological examination of buffalos

A revisão tem como objetivo reavivar as particularidades da espécie bubalina visando a avaliaçãoandrológica. São abordadas as particularidades e anormalidades de desenvolvimento do sistema genital domacho, aspectos da puberdade e maturidade sexual, particularidades do comportamento reprodutivo,características do espermatozoide, liquido seminal e do ejaculado de búfalos.(AU)

The literature review focuses particularities of the buffalo specie and is directed to veterinarians aimingthe breeding soundness evaluation of bulls. Anatomical and abnormal developmental aspects of the genital tractare approached; puberty and sexual maturity, particularities of the reproductive behavior, characteristics of thesperm cells, seminal fluid and ejaculate are discussed.(AU)

Exame andrológico de bubalinos / Andrological examination of buffalos

A revisão tem como objetivo reavivar as particularidades da espécie bubalina visando a avaliaçãoandrológica. São abordadas as particularidades e anormalidades de desenvolvimento do sistema genital domacho, aspectos da puberdade e maturidade sexual, particularidades do comportamento reprodutivo,características do espermatozoide, liquido seminal e do ejaculado de búfalos.

The literature review focuses particularities of the buffalo specie and is directed to veterinarians aimingthe breeding soundness evaluation of bulls. Anatomical and abnormal developmental aspects of the genital tractare approached; puberty and sexual maturity, particularities of the reproductive behavior, characteristics of thesperm cells, seminal fluid and ejaculate are discussed.

The epididymis and its role on sperm quality and male fertility

The epididymis is a complex organ where spermatozoa acquire motility and ability to fertilize theegg. Epididymal maturation lasts 1 or 2 weeks and exposes the immature spermatozoa to a sequentially modified milieu, promoting intense interactions with secretions by the epididymal epithelium. Sperm surface modifications in response to interactions with epididymal secretions are key steps to achieve fertility ability. However, the precise molecular mechanisms that convert an immotile and infertile gamete into a highly motile cellcapable of fusion with an oocyte are still unknown. Recent data on proteomics and transcriptomics of epididymal fluid and epididymosomes brought new ideas of the physiological roles of proteins and miRNAs in epididymal maturation in spermatozoa. This review focuses on the recent discoveries on epididymal fluid composition and its role on sperm maturation and preservation, linking to their survival and fertility potential.

The epididymis and its role on sperm quality and male fertility

The epididymis is a complex organ where spermatozoa acquire motility and ability to fertilize theegg. Epididymal maturation lasts 1 or 2 weeks and exposes the immature spermatozoa to a sequentially modified milieu, promoting intense interactions with secretions by the epididymal epithelium. Sperm surface modifications in response to interactions with epididymal secretions are key steps to achieve fertility ability. However, the precise molecular mechanisms that convert an immotile and infertile gamete into a highly motile cellcapable of fusion with an oocyte are still unknown. Recent data on proteomics and transcriptomics of epididymal fluid and epididymosomes brought new ideas of the physiological roles of proteins and miRNAs in epididymal maturation in spermatozoa. This review focuses on the recent discoveries on epididymal fluid composition and its role on sperm maturation and preservation, linking to their survival and fertility potential.(AU)

Condição metabólica e integridade das membranas, plasmática e acrossomal, da célula espermática bovina após criopreservação em diferentes meios diluidores acrescidos com diferentes tipos de insulina / Metabolic condition and integrity of membranes, and plasma acrosomal, cell bovine sperm after cryopreservation in different extenders greater with different kinds of insulin

Seminal plasma is a fluid with essential role in sperm functions in vivo, from ejaculation to fertilization.Among the hormones present in this medium, insulin stands out, due to its key role in the structure and motility,favoring fertilization. Insulin acts as preservation factor not able sperm and may be the crux of spermpreservation when using cryogenic processes. This study aimed to evaluate the metabolic and structuralcondition of bovine sperm cells after freezing using extenders plus different types of insulin and different types ofegg yolk. Our results showed that sperm motility was not affected by treatments used (insulin and yolk). Inconclusion, the type of egg yolk, the paw (Anas platyrhynchos), increased the viability of cryopreserved spermcells, regardless of the type of insulin used.(AU)

Condição metabólica e integridade das membranas, plasmática e acrossomal, da célula espermática bovina após criopreservação em diferentes meios diluidores acrescidos com diferentes tipos de insulina / Metabolic condition and integrity of membranes, and plasma acrosomal, cell bovine sperm after cryopreservation in different extenders greater with different kinds of insulin

Seminal plasma is a fluid with essential role in sperm functions in vivo, from ejaculation to fertilization.Among the hormones present in this medium, insulin stands out, due to its key role in the structure and motility,favoring fertilization. Insulin acts as preservation factor not able sperm and may be the crux of spermpreservation when using cryogenic processes. This study aimed to evaluate the metabolic and structuralcondition of bovine sperm cells after freezing using extenders plus different types of insulin and different types ofegg yolk. Our results showed that sperm motility was not affected by treatments used (insulin and yolk). Inconclusion, the type of egg yolk, the paw (Anas platyrhynchos), increased the viability of cryopreserved spermcells, regardless of the type of insulin used.

A study on vas deference and seminal vesicle of male goats (Capra hircus)

Background: Reproductive activity of male animal is strongly associated with development of the testicle, the conductor channels and accessory sex gland. Vas deference may be considered the extra-testicular continuation from cauda of the epididymis, and it is the portion of the reproductive tract fundamentally associated with transportation of the sperm-containing fluid from each epididymis to the urethra for their finishing discharge. Non-ampulated part is further divided into: extra-abdominal portion along the caudal border of testes, up to the vaginal ring and abdominal portion started from the vaginal ring to ampullae. Accessory sex glands including vesicular gland and ampullae of vas deferens are characteristically essential for reproductive process. Gross morphometric features of anatomical structures of male reproductive tract in an animal make available a very valuable mechanism in understanding of several physiological and reproductive phenomena. The current study aim at documenting baseline data on gross morphometric aspects of vas deference and seminal vesicle in adult male local nondescript goats in Pakistan ecology that could be utilized as reference values in evaluating their congenital defects and gross pathological abnormalities.Materials, Methods & Results: Experiments were carried out on ampullated and non-ampullated segment of vas deference and seminal vesicle. A total of n = 100 local male goats of 2-3 years of age were selected for this study. Vas deference and seminal vesicles (accessory sex gland) were collected immediately after slaughter from local abattoirs. The non-ampullated segment of vas deference was further divided into abdominal and extra-abdominal segment. Specimens were dissected and washed with normal saline. Standard procedure was adopted for these morphometric features of vas deference and seminal vesicle by using vernier caliper, scale, non-stretchable thread and electronic weighing balance.[...](AU)

A study on vas deference and seminal vesicle of male goats (Capra hircus)

Background: Reproductive activity of male animal is strongly associated with development of the testicle, the conductor channels and accessory sex gland. Vas deference may be considered the extra-testicular continuation from cauda of the epididymis, and it is the portion of the reproductive tract fundamentally associated with transportation of the sperm-containing fluid from each epididymis to the urethra for their finishing discharge. Non-ampulated part is further divided into: extra-abdominal portion along the caudal border of testes, up to the vaginal ring and abdominal portion started from the vaginal ring to ampullae. Accessory sex glands including vesicular gland and ampullae of vas deferens are characteristically essential for reproductive process. Gross morphometric features of anatomical structures of male reproductive tract in an animal make available a very valuable mechanism in understanding of several physiological and reproductive phenomena. The current study aim at documenting baseline data on gross morphometric aspects of vas deference and seminal vesicle in adult male local nondescript goats in Pakistan ecology that could be utilized as reference values in evaluating their congenital defects and gross pathological abnormalities.Materials, Methods & Results: Experiments were carried out on ampullated and non-ampullated segment of vas deference and seminal vesicle. A total of n = 100 local male goats of 2-3 years of age were selected for this study. Vas deference and seminal vesicles (accessory sex gland) were collected immediately after slaughter from local abattoirs. The non-ampullated segment of vas deference was further divided into abdominal and extra-abdominal segment. Specimens were dissected and washed with normal saline. Standard procedure was adopted for these morphometric features of vas deference and seminal vesicle by using vernier caliper, scale, non-stretchable thread and electronic weighing balance.[...]

Perfil proteico do sêmen de cães: em busca de um antígeno espermático para contracepção

Vacinas contraceptivas têm sido propostas como uma forma mais rápida e menos invasiva para esterilização de cães. Este estudo objetivou descrever a relação entre as proteínas das células espermáticas ejaculadas e do epidídimo, do plasma seminal e do fluído epididimário, produzindo base experimental para o design destas vacinas. Para isso, utilizou-se 5 cães (n = 5), dos quais foram obtidos o ejaculado e, após a orquiectomia, o produto da recuperação da cauda do epidídimo. As amostras foram submetidas a avaliação da concentração, morfologia, cinética espermática, citometria de fluxo e teste de ligação em membrana perivitelínica de ovo, sendo separadas em 4 grupos: plasma seminal (PS), fluido epididimário (FE), espermatozoides do ejaculado (EJAC) e do epidídimo (EPID). Para análise proteômica a digestão das proteínas dos 4 grupos foi feita in gel e avaliadas em espectrômetro de massas Q-Tof. Os parâmetros de motilidade progressiva, retilinearidade, linearidade, coeficiente de oscilação, produção de H2O2 e percentagem de membrana plasmática íntegra e acrossomal lesada foram mais expressivos (P< 0,01) no grupo EJAC do que no EPID. Foram encontradas 177 proteínas entre EJAC e PS (16 up-regulated no EJAC e 14 no PS); 241 proteínas entre EPID e FE (27 up-regulated no EPID e 14 no FE); e 210 proteínas entre EJAC e EPID (3 up-regulated no EJAC e 17 no EPID).

Contraceptive vaccines has been proposed as a faster and less invasive way to sterilize dogs. This study aimed to describe the relation between proteins of ejaculated sperm cells and epididymis, seminal plasma and epididymal fluid, producing an experimental basis for designing these vaccines. For this, 5 dogs (n = 5) were used, from which the semen and, after orchiectomy, the epididymis tail recovery product was obtained. Samples were submitted to concentration, morphology, sperm kinetics, flow cytometry and egg perivitellin membrane binding test analisys, and separated into 4 groups: seminal plasma (PS), epididymal fluid (EF), ejaculate spermatozoa (EJAC) and epididymis spermatozoa (EPID). For proteomic analysis the protein digestion of the 4 groups was performed in gel and evaluated by Q-Tof mass spectrometer. The parameters of progressive motility, rectilinearity, linearity, wobble, H2O2 production and percentage of plasma membrane integrity and damaged acrosomal membrane were more expressive (p <0.01) in the EJAC group than in EPID group. 177 proteins were found between EJAC and PS (16 up-regulated in EJAC and 14 in PS); 241 proteins between EPID and FE (27 up-regulated in EPID and 14 in FE); and 210 proteins between EJAC and EPID (3 up-regulated in EJAC and 17 in EPID).

Sistema reprodutor masculino em Porcellanidae: reconstrução do caráter ancestral com base na morfologia, histoquímica e ultraestrutura dos espermatóforos e espermatozoides.

Os caranguejos da família Porcellanidae tem seu sistema reprodutor masculino desconhecido do ponto de vista histológico e pouco estudado no aspecto ultraestrutural dos espermatozoides. Nesta dissertação, apresentamos o padrão anato-histológico do sistema reprodutor masculino, além da ultraestrutura dos espermatóforos e espermatozoides de Porcellanidae, buscando entender as relações de parentesco e a evolução do sistema reprodutor e suas células germinativas entre as superfamílias da infraordem Anomura. Foram coletados com auxílio de um barco de pesca camaroeira, além de coleta manual na região entre marés e por meio de mergulho livre e autônomo (3m), machos adultos dos gêneros Pachycheles, Petrolisthes, Pisidia, Porcellana e Megalobrachium em Ubatuba e São Vicente (SP), Brasil, e em Valdivia, Chile. As amostras foram processadas seguindo rotinas para histologia, microscopia eletrônica de trasmissão e varredura. Análises do fluido seminal, empacotamento dos espermatozoides e formação dos espermatóforos, apresentaram o mesmo padrão para todas as espécies estudadas, e semelhante a outros Anomuras já descritos na literatura, sendo simplificado e sem diferenciações macroscopícas. Os espermatóforos de Porcellanidae seguem o padrão tripartido pedunculado como descrito para a maioria dos Anomura, porém se diferenciam por apresentarem um pedúnculo diminuto, característica compartilhada com Galatheidae e Munidae. A ultraestrutura dos espermatozoides dos Porcellanidae é caracterizada por uma vesícula acrossomal saliente ao espermatozoide, possuindo camadas concêntricas e diferente dos outros Galatheoidea por apresentarem: opérculo centralmente perfurado, cone perforatorial eletrondenso, e anel perforatorial posterior. Porcellanidae possuem caracteristicas compartilhadas entre as espécies mais proximamente relacionadas e algumas exclusivas de cada gênero. Pisidia brasiliensis e Porcellana sayana, possuem um ombro na região apical do acrossoma. Porcellana sayana e Megalobrachium roseum o cone perforatorial se estende até a abertura opercular. No gênero Petrolisthes, ocorre uma estrutura excusiva, o anel tubular eletrondenso. Pisidia brasiliensis possui botão apical. Assim, a ultraestrutura do espermatozoide destas espécies de Porcellanidae representam um novo conjunto de caracteres os quais são claramente relacionados filogeneticamente, quando traçados por meio do uso da filogenia mais recente da infraordem.

The crabs of the Porcellanidae family have their male reproductive system unknown from the historical point of view and little studied in the ultrastructural aspect of sperm. In this dissertation, we present the histological pattern of the male reproductive system, in addition to the ultrastructure of sperm and sperm from porcellanids, seeking to understand how kinship relationships and the evolution of the reproductive system and its germ cells between the superfamilies of Anomura's infrastructure. Were collected with the aid of a camaro fishing boat, in addition to manual collection in the region between tides and free and autonomous diving medium (3m), adult males of the genera Pachycheles, petrolitic, pisis, porcellana and megalobrachium in Ubatuba and São Vicente (SP ), Brazil, and in Valdivia, Chile. As the samples were processed following routines for histology, transmission electron microscopy and scanning. Analysis of seminal fluid, sperm packaging and sperm formation, analysis or the same pattern for all studied species, and similar to other Anomuras already applied in the literature, being simplified and without macroscopic differences. Porcellanid spermatozoa follow the pedunculated tripartite pattern as described for most Anomura, but they differ by presenting a tiny peduncle, resources shared with Galatheidae and Munidae. An ultrastructure of porcelain spermatozoa is characterized by an acrosomal vesicle protruding from the sperm, having concentric layers and different from the other Galatheoidea in that they have: centrally perforated operculum, perforated electron- condensed cone and perforated posterior ring. Porcellanidae has characteristics shared between the closest species and some exclusive to each genus. Pisidia brasiliensis and Porcellana sayana, have a "shoulder" in the apical region of the acrosome. Porcellana sayana and Megalobrachium roseum or perforated cone extend to the opercular opening. In the Petrolisthes genus, there is an excusive structure or an electronic tubular ring. Pisidia brasiliensis has an apical bud. Thus, an ultrastructure of the sperm of these species of Porcellanidae represents a new set of characters, which are clearly related phylogenetically, when traced using the most recent phylogeny of the infraorder.