Influência de diferentes sistemas e curvas de congelamento na congelabilidade e fertilidade do sêmen equino / Influence of systems and freezing curve in equine semen freezability and fertility

Avaliou-se o efeito de curvas de congelação nos parâmetros espermáticos e na fertilidade, usando sêmen de alta e baixa congelabilidade. Experimento 1 - utilizou-se sêmen de quatro garanhões resistentes à congelação: grupo 1, palhetas refrigeradas até 5°C e congeladas com curva de -8°C/min; grupos 2 e 3, palhetas refrigeradas até 5°C (0,5°C/min.) e congeladas com curvas de -20°C/min e -10°C/min, respectivamente. Experimentos 2 e 3 - utilizaram-se cinco garanhões (Mangalarga Marchador), respectivamente, de alta e baixa congelabilidade: grupo 4, a mesma metodologia descrita no grupo 1; grupos 5 e 6, palhetas refrigeradas até 5°C (0,5°C/min) e congeladas com curva de -20°C/min, entre 5°C e -60°C, e -10°C/min, entre -60°C e -100ºC (grupo 5), e -25°C/min, de 5°C até -100°C (grupo 6). O sêmen foi avaliado após descongelamento pelo método computadorizado. No experimento 1, não houve diferença nos parâmetros avaliados. No experimento 2, os parâmetros motilidade total (MT) e motilidade progressiva foram superiores aos do grupo 6 em relação ao grupo 4. No experimento 3, a MT foi superior no grupo 6 em relação ao grupo 4. As curvas de congelação mais rápidas apresentaram melhores parâmetros de cinética espermática, após a descongelação, para o sêmen de garanhões da raça Mangalarga Marchador.(AU)

The effect of freezing curves on sperm parameters and fertility, using resistant and sensitive semen to cryopreservation, was evaluated. In experiment 1, Semen from 4 stallions resistant to freezing was used: Group 1, straws were cooled to 5°C and frozen with a curve of - 8°C/min; Groups 2 and 3, straws were cooled to 5°C (0.5°C/min) and frozen with curves of - 20°C / min and - 10°C/min, respectively. In experiments 2 and 3, 5 stallions (Mangalarga Marchador) presenting respectively resistant and sensitive sperm to cryopreservation were used: Group 4, same methodology described for Group 1 was performed; Groups 5 and 6, straws were cooled to 5°C (0.5°C/min) and frozen with a curve of - 20°C/min. between 5°C and - 60°C and -10°C/min. between - 60°C and - 100°C (Group 5) and - 25°C/min. 5°C to - 100°C (Group 6). Thawed-semen was evaluated by the computerized method CASA. In Experiment 1, there was no difference in the evaluated parameters. In Experiment 2, total motility (MT) and progressive motility (PM) were higher in Group 6 compared to Group 4. In Experiment 3, TM was higher in Group 6 than Group 4. The faster freezing curves showed better parameters of sperm kinetics after thawing, for the Mangalarga Marchador stallion semen.(AU)

Development of Sperm Separation System using Electrical Current for Bull / Jurnal Sains Kesihatan Malaysia

@#A novel electrophoretic separation system has been successfully applied for the preparation of human sperm prior to the execution of assisted reproductive techniques (ARTs). This new system is designed to overcome the generation of reactive oxygen species (ROS) through centrifugation in conventional sperm preparation. Since the previous study showed favorable outcomes in humans, this study intends to implement this new system for animal sperm preparation particularly in bull. Fresh semen from adult bulls were used. Optimization of the electrophoretic system for optimum bull sperm separation involved different strength of voltage and separation time. The voltages applied were 10V, 20V, 30V, 40V, 50V, and 60V. For each voltage applied, the system was operated for a duration of 12 min. An average of 10 µl fractionalized semen was taken out at the collection site at every 2-min interval. Every fractionated sperm was then evaluated for percentage of viability, motility, and DNA damage assessment. Result showed that electrophoresis at 20V and 6 min yielded more than 80% viable and more than 70% motile sperm population with the lowest DNA damage. In conclusion, the system was able to fractionate high quality bull sperm at 20V and 6 min.

Cryopreservation of boar semen in 0. 5 mL straws at low spermatozoa concentration is better than high concentration to maintain sperm viability / A manutenção da viabilidade do sêmen suíno criopreservado em palhetas de 0, 5 mL em baixas concentrações espermáticas é melhor do que em altas concentrações

To date, no studies have been performed evaluating the effect of boar spermatozoa concentration in 0.5mL freezing straws, leading us to examine this question. Each sperm-rich fraction of the ejaculate (n=25) was diluted at five different sperm concentrations (100, 200, 300, 600 and 800 x 106 spermatozoa/mL), packaged in 0.5mL straws, and subsequently frozen. After thawing, the sperm from all of treatment groups were analyzed to determine motility characteristics using a sperm class analyzer (SCA-CASA), and their plasma and acrosomal membrane integrity, mitochondrial membrane potential, sperm membrane lipid peroxidation and fluidity were analyzed by flow cytometry. An increase in spermatozoa concentration above 300x106 spermatozoa/mL in a 0.5mL straw impaired (p<0.05) the total and progressive motility, curvilinear velocity, straight-line velocity, linearity and beat cross frequency. However, the plasma and acrosomal membrane integrity, mitochondrial membrane potential, membrane lipid peroxidation and fluidity were not influenced (p>0.05) by high spermatozoa concentrations at freezing. Therefore, to increase spermatozoa survival and total and progressive motility after thawing, boar spermatozoa should be frozen at concentrations up to 300x106 spermatozoa/mL.(AU)

Até o momento, não foram realizados estudos que avaliassem o efeito da concentração de espermatozoides/mL em palhetas (0,5mL) para a criopreservação, levando-nos a analisar esta questão. Cada fração-rica do ejaculado (n=25) foi diluída em cinco diferentes concentrações de espermatozoides (100, 200, 300, 600 e 800x106 espermatozoides/mL), envasadas em palhetas de 0,5mL e posteriormente congeladas. Após a descongelação, os espermatozoides de todos os tratamentos foram avaliados a fim de determinar as características de motilidade usando um sistema de análise computadorizada dos espermatozoides (SCA-CASA). A integridade das membranas plasmática e acrosomal, o potencial de membrana mitocondrial, a peroxidação lipídica e a fluidez da membrana foram analisadas por citometria de fluxo. O aumento na concentração de espermatozoides acima de 300x106 espermatozoides/mL diminuiu (p<0,05) a motilidade total e progressiva, velocidade curvilínea, velocidade linear, linearidade e frequência de batimento. No entanto, a integridade da membrana plasmática e acrosomal, potencial de membrana mitocondrial, peroxidação lipídica e fluidez de membrana não foram influenciados (p>0,05) por altas concentrações de espermatozoides durante a criopreservação. Portanto, a fim de melhorar a sobrevivência dos espermatozoides suínos e a motilidade total e progressiva após a descongelação, os espermatozoides suínos devem ser congelados a concentrações não superiores a 300x106 espermatozoides/mL.(AU)

Correlation between the level of presenilin-associated rhomboid-like protein and the sperm motility, survival rate and deformity rate of sperm in workers exposed to aluminum in Guangxi / 国际检验医学杂志

Objective To investigate the correlation between the level of presenilin-associated rhomboidlike protein (PARL) and the sperm motility,survival rate and deformity rate of sperm in workers exposed to aluminum.Methods A total of 162 male workers exposed to aluminum in a large aluminum enterprise in Guangxi were selected as the exposure group,and the 162 staff members of the service company were selected as the control group by matching the age and length of service.The concentration of aluminum in the working environment and the aluminum content in the blood and urine were determined by graphite furnace atomic absorption spectrometry and high performance liquid chromatography.Enzyme linked immunosorbent assay was used to measure the level of PARL protein in sperm and sperm function was evaluated.Results The average concentrations of aluminum in the batching,electrolysis and casting of aluminum exposure group were (6.72± 1.45),(7.23± 1.50) and (7.35± 1.72)mg/m3,which were significantly higher than those of the control group (F=8.314,P＜0.001).The content of aluminum in blood and urine of exposed group were significantly higher than those of control group (P＜0.05),while the levels of PARL of exposed group were significantly lower than those of control group (P＜0.05).Moreover,the sperm survival rate and sperm motility of exposed group were significantly lower than those of the control group,and the sperm deformity rates of exposed group were significantly higher than those of the control group(P＜0.05).Pearson correlation analysis showed that there was a positive correlation of sperm motility,sperm survival rate and protein level of PARL (rsprm motility=0.713,P=0.012;rsperm survival rate =0.628,P=0.008);while the sperm deformity rate and protein level of PARL showed a significant negative correlation (rsperm deformity rate =0.953,P =0.002).Conclusion The sperm function was significantly impaired in aluminum exposed workers,and the changes of sperm motility,survival rate and malformation rate were closely related to the protein level of PARL.

Yam polysaccharide improves sperm viability and protects sperm DNA integrity in vitro / 中华男科学杂志

Objective@#To clarify the roles of yam polysaccharide (YPS) in improving sperm viability and protecting sperm DNA integrity in vitro and provide a new approach to the treatment of oligoasthenozoospermia.@*METHODS@#We collected samples by masturbation from 36 normal fertile males aged 27－39 years. Each sample was divided into six groups: blank control or treated with normal saline, vitamin C solution, and YPS solution at low (0.25 mg/ml), medium (1.0 mg/ml) or high concentration (5.0 mg/ml). Using eosin-Y staining, sperm hypotonic swelling (HOS) and sperm chromatin diffusion (SCD) test, we observed the effects of different concentrations of YPS on sperm viability, membrane integrity and nuclear DNA.@*RESULTS@#After 24 and 48 hours of treatment, sperm viability was markedly reduced in the vitamin C (［28.5 ± 3.1］ and ［6.5 ± 1.2］%), low-YPS (［31.3 ± 3.5］ and ［6.5 ± 2.2］%), medium-YPS (［37.1 ± 3.5］ and ［9.5 ± 2.8］%) and high-YPS groups (［38.3 ± 3.3］ and ［9.0 ± 3.2］%) as compared with the blank control (［17.3 ± 2.1］ and ［3.2 ± 1.3］%) (P 0.05).@*CONCLUSIONS@#Yam polysaccharide can improve sperm viability and protect sperm DNA integrity in vitro.

Efecto in vitro del pentóxido de vanadio (V2O5) sobre la calidad espermática / In vitro effect of vanadium pentoxide (V2O5) on sperm quality

El vanadio es un metal pesado considerado como un contaminante ambiental, producto de la manipulación antropogénica, que incide sobre el potencial reproductivo masculino, especialmente sobre la movilidad espermática. Esta investigación tuvo como objetivo determinar el efecto in vitro del pentóxido de vanadio sobre la calidad espermática. Se incubaron suspensiones de espermatozoides humanos por 24 horas bajo concentraciones de 0, 1 y 2 ppm V2O5, se realizó la valoración de la movilidad y vitalidad espermática, la integridad de membrana, la reacción acrosómica y la integridad de la cromatina espermática. El V2O5 alteró de manera significativa la movilidad espermática, específicamente los patrones de movilidad espermática moderado (p < 0.01), lento (p = 0.01) e inmóvil (p < 0.01). Los grupos tratados con V2O5 presentaron un porcentaje de espermatozoides vivos significativamente mayor al grupo control (p < 0.01). La integridad de membrana y reacción acrosómica no resultaron afectadas por la exposición de espermatozoides humanos a V2O5 (p > 0.05). De igual modo, no se observaron alteraciones del material nuclear (p > 0.05). El vanadio es capaz de alterar la movilidad y vitalidad espermática sin inducir cambios sobre la integridad de membrana y la cromatina espermática.

Vanadium is a heavy metal considered an environmental pollutant product of anthropogenic manipulation; it influences the masculine reproductive potential, especially the sperm motility. This research had the objective of determining the effect of vanadium pentoxide on sperm quality in vitro. Spermatozoa were incubated for 24 hours under 0, 1 and 2 ppm V2O5 concentrations, and sperm motility and vitality, membrane integrity, acrosome reaction and sperm chromatin integrity were assessed. V2O5 altered sperm motility in a significant way, specifically moderated (p < 0.01), slow (p = 0.01), and non motile (p < 0.01) sperm motility patterns. The groups treated with V2O5 had a major percentage of live spermatozoa than the control group (p < 0.01). Membrane integrity and acrosome reaction were not affected by human sperm exposition to V2O5 (p > 0.05). Similarly, we did not observe nuclear material alterations. Vanadium is able to alter sperm motility and viability without inducing changes on membrane and chromatin integrity.

Dois diluentes comerciais na criopreservação de espermatozoides do epidídimo de touros / Two commercial extenders for cryopreservation of epididymal bull sperm

A recuperação e a criopreservação de espermatozoides do epidídimo constituem alternativas viáveis para a preservação de material genético de animais valiosos. O objetivo deste estudo foi comparar o desempenho de dois diluentes comerciais Botu-Bov(r) (BB) e Bovimix(r) (BV), sobre a viabilidade pós-descongelação de espermatozoides do epidídimo de touros Tabapuã (Bos taurus indicus) pós-castração. Os espermatozoides foram colhidos da cauda de 20 epidídimos utilizando a técnica de fluxo retrógrado, centrifugados e diluídos com BB ou BV para posterior criopreservação a -196°C. Após a descongelação, as amostras foram avaliadas utilizando a análise computadorizada (CASA) e por análises microscópicas para a determinação da integridade de membranas plasmáticas, acrossomal e morfologia espermática. A avaliação estatística dos dados foi realizada pela análise de variância (ANOVA) com o pós-teste de comparações múltiplas de Tukey-Kramer, com nível de significância (P<0,05). Os resultados do movimento espermático avaliado pelo CASA, não diferiram para o diluente BB e BV. Também não foi observada diferença significativa entre os grupos no percentual de espermatozoides morfologicamente deformados, defeitos de acrossoma e espermatozoides com membrana plasmática íntegra após o descongelamento. Conclui-se que ambos os diluentes (BB e BV) são eficientes e podem ser utilizados na tecnologia do congelamento de espermatozoides colhidos da cauda do epidídimo de touros, não apresentando diferença na viabilidade espermática para os parâmetros estudados.

Recovery and cryopreservation of epididymal sperm is a viable alternative for preservation of genetically valuable animals. The aim of this study was to verify and to compare the effect of two commercial extenders for conventional semen on post-thawing viability of bovine epididymal sperm. For this purpose, the spermatozoa was recovered from the tail of 20 epididymis of Tabapuã bulls (Bos Taurus indicus) using retrograde flow method. After sperm recovery, the cells were centrifuged and divided for dilution with the diluents Botu-Bov(r) (BB) or Bovimix(r) (BV) for cryopreservation at -196°C. After thawing, all samples were evaluated using computer assisted sperm analysis (CASA), and by microscopic analysis for determination of integrity of plasma and acrossomal membrane and morphology. Statistical evaluation was performed by analysis of variance (ANOVA) with post-test for multiple comparisons, the Tukey-Kramer test, with significance level (P<0.05). The results of the sperm movement for diluent BB and BV evaluated with CASA, showed no difference for both (P>0.05). There was also no difference between the percentage of deformed sperm, acrosome defects and the sperm with intact plasma membrane after thawing with BB or BV. We conclude that both extenders (BB and BV) are efficient and can be used for freezing sperm collected from the epididymis of bulls, showing no difference for all the parameters studied.

Spermatozoa characterization in the one-sided livebearing Jenynsia multidentata (Cyprinodontiformes: Anablepidae) / Caracterización de parámetros espermáticos del pez vivíparo Jenynsia multidentata (Cyprinodontiformes: Anablepidae)

Several sperm parameters have been employed as useful tools to evaluate fish fertility. Within teleosts, approximately 3% of fish species are known to be viviparous. The Order Cyprinodontiformes includes several species with internal fertilization, and within this group most of the studies about sperm quality have been mainly focused on the Poeciliidae family. The livebearing fish Jenynsia multidentata (Anablepidae) inhabits an extensive area of the Neotropical region and it has been used as a useful fish laboratory model to evaluate the effects of xenobiotics through different biomarkers. The present work characterized the sperm of this species through a simple protocol of semen collection. Sperm population showed linearity greater than 89% and 70% of fish have a straight line and curvilinear velocity valued between 50 and 100µm/s. Although 85% of individuals showed a proportion of live sperm higher than 60%, the male population had a high degree of heterogeneity in its sperm count. Morphometry analyses showed a total sperm and head lengths of 46.66±2.06µm and 3.46±0.41mm, respectively. A rather long midpiece region (9.12±0.65µm) was registered, which may indicate high energy-producing capabilities of the spermatozoa. This study established basic parameter values which could be useful for evaluating reproductive potential of J. multidentata populations.

Diversos parámetros espermáticos han sido utilizados para evaluar la fertilidad de peces. Dentro de los peces teleósteos, aproximadamente el 3% de las especies son vivíparas. El orden Cyprinodontiformes incluye varias especies con fecundación interna. Dentro de este orden la mayor parte de los estudios sobre la calidad del esperma se han centrado principalmente en la familia Poeciliidae. El pez vivíparo Jenynsia multidentata (Anablepidae) habita una extensa área de la región Neotropical y ha sido utilizado como un exitoso modelo de laboratorio. El objetivo del presente trabajo fue caracterizar los espermatozoides de esta especie a través de un simple protocolo de recolección de esperma. La población de espermatozoides mostró una linealidad superior al 89% y el 70% de los peces tienen una velocidad lineal y curvilineal entre 50 y 100µm/s. Aunque el 85% de los individuos mostró una proporción de espermatozoides vivos de más del 60%, se observó una alta heterogeneidad en el recuento espermático. Los análisis morfométricos mostraron una longitud total de espermatozoides de 46.66±2.06µm y una longitud de la cabeza de 3.46±0.41µm. Los espermatozoides presentan una pieza media larga (9.12±0.65µm) lo que puede indicar una alta capacidad de producción de energía. El presente estudio establece valores básicos de parámetros que pueden ser útiles para evaluar el potencial reproductivo de las poblaciones de J. multidentata.

La calidad espermática evaluada mediante metodologías no convencionales / Sperm quality analysis by non-conventional methodologies

Existen diferentes metodologías para evaluar la calidad seminal, siendo la valoración de la movilidad y de la morfología espermática los indicadores más comúnmente utilizados, sin embargo, los espermatozoides poseen ciertas características que no siempre pueden analizarse a través del examen tradicional. En esta revisión de la literatura se describen algunas metodologías alternativas empleadaspara observar y evaluar las características seminales. La movilidad, la viabilidad y la morfología espermática pueden evaluarse empleando metodologías manuales y análisis asistidos por computador. Otras características evaluables de la biología espermática son la producción de especies reactivas del oxígeno, la calidad mitocondrial y el ADN espermático. Esta revisión demuestra que existe una amplia disponibilidad de metodologías para el análisis seminal, sin embargo, cada día se siguen implementando nuevas técnicas, lo que impactará en el entendimiento de la fisiología espermática. En un futuro estas herramientas diagnósticas podrán incidir en el beneficio de los pacientes con infertilidad.

There are different methodologies for assessing semen quality assessment of mobility and sperm morphology are the most commonly used indicators. However, sperm have certain characteristics that cannot always be analyze through the traditional examination. In this review are describe some alternativemethodologies to observe and assess the seminal characteristics. Motility, viability and sperm morphology can be evaluated using manual methodologies and computational analysis. Other quantifiable characteristics of sperm biology are the production of reactive oxygen species, mitochondrial and DNA sperm quality. Here is shown that there are many methodologies for seminal analysis, however, each day are going to implementing new techniques, which will impact on the understanding of sperm physiology and in the future, they may improve the diagnosis of individuals.

Cooling of ejaculated and epididymal stallion sperm / Refrigeração de espermatozoides do ejaculado e epidídimo em garanhões

After a serious injury or sudden death, epididymis cauda sperm recovery and cryopreservation may present as the last opportunity to obtain genetic material from a valuable stallion. This study evaluated the viability of cooled equine sperm collected by three different methods: sperm of ejaculated (G1), sperm recovered from the epididymal cauda immediately after orchiectomy (G2) and sperm recovered from the epididymal cauda after storage for 24 hours at 5°C (G3). To obtain G1 sperm, two ejaculates were collected. After 1 week, all stallions underwent a bilateral orchiectomy, and one of the removed epididymides was flushed to obtain G2 sperm. The contralateral epididymis was stored at 5°C for 24 hours before being flushed to obtain G3 sperm. The sperm samples were evaluated immediately after the addition of the refrigeration extender, and after 24 and 48 hours of storage at 5°C. After 24 and 48 hours of storage, the epididymal sperm demonstrated higher motility traits when compared to the ejaculated sperm (P<0.05). These results indicate that sperm recovered from the epididymal cauda of stallions are more resistant to the cooling process, with higher kinetic parameters and plasma membrane integrity when compared to the ejaculated sperm.

A recuperação de espermatozoides da cauda do epidídimo pode ser a última chance para preservação do germoplasma quando ocorre morte súbita ou lesão grave em garanhões de alto valor genético. O presente trabalho comparou a viabilidade após refrigeração dos espermatozoides do ejaculado (G1), recuperados da cauda do epidídimo imediatamente após a orquiectomia (G2) e recuperados após armazenamento do epidídimo por 24 horas a 5ºC (G3). No G1 foram colhidos dois ejaculados. Uma semana após a colheita dos ejaculados os garanhões foram submetidos à orquiectomia bilateral e realizada a colheita dos espermatozoides da cauda do epidídimo de um testículo de cada garanhão (G2). O testículo contralateral permaneceu a 5°C por 24 horas, antes da recuperação espermática (G3). A análise das amostras foi realizada imediatamente após a adição do meio de refrigeração, e após 24 e 48 horas de armazenamento a 5°C. Após 24 e 48 horas de armazenamento, os espermatozoides do epidídimo demonstraram características de cinética maiores que os do ejaculado (P<0.05). Estes resultados indicam que espermatozoides recuperados da cauda do epidídimo foram mais resistentes ao processo de refrigeração, com maiores parâmetros de cinética espermática e integridade da membrana plasmática quando comparados aos espermatozoides do ejaculado.

Espermatozoides caprinos criopreservados em meio à base de leite desnatado acrescido de glutationa reduzida / Goat spermatozoa after freezing in skimmed-milk extender supplemented with reduced glutathione

Visando avaliar o efeito da adição de glutationa reduzida (GSH) ao diluente de congelação de sêmen caprino à base de leite desnatado, utilizou-se sêmen de cinco reprodutores Boer. Após colheita e avaliação, procedeu-se à formação do pool dos ejaculados e diluição em leite desnatado e glicerol 7 por cento, acrescido de antioxidantes: G1) Controle; G2) GSH 2mM mL-1; G3) GSH 5mM mL-1 e G4) GSH 7mM mL-1. As amostras foram congeladas em palhetas (0,25mL) e armazenadas a -196°C. Após descongelação, avaliou-se a integridade de membrana plasmática (iMP) e acrossomal (iAc), potencial de membrana mitocondrial (PMM), cinética e ultraestrutura. Os grupos Controle e GSH (2, 5 e 7mM mL-1) não diferiram (P>0,05) em iMP, iAc, PMM e cinética. Na análise ultraestrutural, os porcentuais de membrana plasmática (cabeça e cauda) e acrossoma íntegros não diferiram (P>0,05) entre grupos. Todavia, o grupo Controle apresentou maior porcentual (P<0,05) de gametas com axonema íntegros do que os de GSH (2, 5 e 7mM mL-1). Maior porcentagem (P<0,05) de espermatozoides com mitocôndrias íntegras foi observada no grupo Controle do que nos de GSH (5 e 7 mM mL-1). Conclui-se que a adição de GSH (2, 5 e 7mM mL-1) em diluente de congelação de sêmen caprino, à base de leite desnatado, não preserva a integridade dos espermatozoides.

Aiming to evaluate in vitro effect of different concentrations of glutathione reduced (GSH) in skimmed-milk and glycerol 7 percent it was used semen from five Boer bucks. After collect and evaluation, a pool of samples was diluted in skimmed-milk and glycerol 7 percent plus antioxidant: G1) Control; G2) GSH 2mM mL-1; G3) GSH 5mM mL-1 and G4) GSH 7mM mL-1. Samples were frozen in straws (0.25mL) and stored at -196°C. After thawing, samples were subjected to integrity of the plasma membrane (iMP) and acrosomal (iAc), mitochondrial membrane potential (MMP), kinematic and ultrastructure analysis. Control and GSH (2, 5 and 7mM mL-1) groups did no differ (P>0.05) in iMP, iAc, PMM and kinematic parameters. In the ultrastructural analysis, percentages of acrosome and plasma membrane (tail and head region) intact did not differ (P>0.05) between groups. However, Control group had higher percentage (P<0.05) of gametes with intact axonemes than those of GSH (2, 5 and 7mM mL-1) groups. Higher percentage (P<0.05) of sperms with intact mitochondrias were observed on Control group than those of GSH (5 and 7mM mL-1). It can be concluded that the GSH (2, 5 and 7mM mL-1) addition in skimmed-milk diluent to freeze goat semen did not preserve sperm integrity.

Análisis de los parámetros del eyaculado en la rata Wistar de laboratorio: descripción de la técnica / Analysis of the parameters of the ejaculate in the laboratory Wistar rat: technical description

The number of spermatozoa, weight and size of the seminal plug has been quantified in the ejaculate of the laboratory rat and sperm viability, sperm mobility and sperm concentration in samples obtained from the epididymis. The reference values for the rat ejaculate have not been determined maybe due to the difficulty to obtain it directly from the male. Nevertheless, an approach can be obtained analyzing the ejaculate collected from the inseminated female. The objective of this study was to evaluate and propose the values of the macroscospic and microscopic parameters of the semen and seminal plug obtained from the female. The seminal analysis of the Wistar rats was performed adapting the methods used for other species. After one ejaculatory series, semen contained in the uterine horns was obtained, as well as the seminal plug from the vagina. In more than one hundred ejaculates, 94.54% were off-white with 2.35 ± 0.06 mm of viscosity and 8.13 ± 0.02 of pH. The sperm concentration was 16.3 ± 0.59 millions of spermatozoa per mL, 0.72 ± 0.01 mobility index, 64.4 ± 0.7% viability and 99.11 ± 0.20% normal morphology. From 99% of seminal plugs, 92.7% were hardened, they weighed 115.63 ± 1.54 mg, measured 12.41 ± 0.13 and 5.31 ± 0.05 mm of length and width respectively, and volume 87.70 ± 1.74 mm3 . In conclusion, the used method to obtain and to evaluate the parameters of the ejaculate is reliable, for that reason, it is suggested that the results obtained could be considered as indicative values for semen and seminal plug for the Wistar rat.

Se ha cuantificado en el eyaculado de la rata de laboratorio, el número de espermatozoides, peso y tamaño del tapón seminal, y en muestras obtenidas de epidídimo, la viabilidad, movilidad y concentración espermáticas. Los valores de referencia para el eyaculado de la rata no se han determinado quizá por la dificultad para obtenerlo directamente del macho. No obstante, puede obtenerse una aproximación al analizar el eyaculado recolectado de una hembra recién inseminada. El objetivo de este estudio fue evaluar y proponer los valores de los parámetros macroscópicos y microscópicos del semen y tapón seminal obtenidos de la hembra. El análisis del semen de ratas Wistar se realizó adecuando los métodos usados para otras especies. Después de una serie eyaculatoria, se obtuvo el semen contenido en los cuernos uterinos y el tapón seminal de la vagina. En más de 100 eyaculados, 94.54% fueron blanquecinos, con 2.35 ± 0.06 mm de viscosidad y 8.13 ± 0.02 de pH. La concentración espermática fue de 16.3 ± 0.59 millones de espermatozoides por mL, 0.72 ± 0.01 de índice de movilidad, 64.4 ± 0.7% de viabilidad y 99.11 ± 0.20% de morfología normal. Del 99% de los tapones seminales, 92.7% fueron endurecidos, pesaron 115.63 ± 1.54 mg, midieron 12.41 ± 0.13 y 5.31 ± 0.05 mm de largo y ancho, respectivamente, y de volumen 87.70 ± 1.74 mm³. En conclusión, el método utilizado para obtener y evaluar los parámetros del eyaculado es confiable, por ello se sugiere que los resultados obtenidos podrían considerarse como valores indicativos para el semen y tapón seminal de la rata Wistar.

Effects of Murine Cytomegalovirus Infection on Sperm Viability in Mice / 华中科技大学学报（医学）（英德文版）

In order to explore the effects of testicular infection of murine cytomegalovirus (MCMV) on mature sperm viability at different periods following MCMV inoculation in mice, 91 BALB/c mice without MCMV infection were randomly divided into two groups: an experimental group (n=56) and a control group (n= 35). The mice in the experimental group were treated by inoculating MCMV intratesticularly, while those in the controlled group were directly inoculated with DMEM without MCMV. The mice in both groups were sacrificed separately on the day 1,1.5, 2, 4, 6, 9 and 14 post-inoculation (D1, 1.5,2, 4, 6, 9 and 14 PI). The MCMV M83 mRNA gene was detected in the testis by in situ hybridization (ISH) with MCMV late-mRNA probe labeled with digoxin.Sperm viability of mature sperm in the epididymis cauda was measured. The results demonstrated the positive signal of ISH of MCMV was found mainly in the cytoplasm of the testicular interstitial cells and spermatogenic cells in the experimental group. Compared with that in the controlled group, the sperm viability in the experimental group was decreased significantly on D1 PI and D1.5PI (P＜ 0.05). No statistically significant difference in the sperm viability was found after D2 PI between two groups (P＞0.05). This suggested that sperm viability in mice might be descended significantly shortly after MCMV infection and might return to normal with time, indicating that MCMV acute infection might temporarily degrade sperm quality and influence procreation transiently.

Relations of sperm morphology with sperm vitality and sperm viability / 吉林大学学报(医学版)

Objective To study the relationships between sperm morphology and sperm vitality,sperm viability.Methods The sperm morphology and the sperm vitality were analyzed by automated sperm morphology analyzer(ASMA) and the sperm viability was analyzed using eosin staining.Results The normal morphology sperm rate of abnormal vitality group was lower than that of normal vitality group(P

Effect of Homologous Human Semen Mixtures on Viability of Sperm / 대한비뇨기과학회지

The causes of male infertility have been well delineated in numerous textbooks and articles. These are divided into six major categories such as failure of spermatogenesis, failure of sperm maturation, failure of sperm transportation, failure of semen composition, failure of hormonal system and failure of ejaculation. Besides these, the motile activity of sperm has been regarded as an important factor for the impregnation This clinical study has been undertaken to examine the effects of hemologous human semen mixtures upon the motile time of the spermatozoa, and the result are presented as follows; Group 1, Mixture of Normospermia and Normospermia: 10 mixtures Normospermia-A: 471. 9 minutes of aver age sperm motile time Normospermia-B: 369. 4 minutes of average sperm motile time Normo-A+Normo-B mixture: 452. 5 minutes of average sperm motile time Group 2, Mixture of Normospermia and Oligospermia: 10 mixtures Normospermia: 445.3 minutes of average sperm motile time Oligospermia: 376.2 minutes of average sperm motile time Normo+Oligo mixture: 456. 1 minutes of average sperm motile time Group 3, Mixture of Normospermia and Azoospermia; 8 mixtures Normospermia: 433. 3 minutes of average sperm motile time Azoospermia: Normo.+Azoo. mixture: 455. 1 minutes of average sperm motile time Group 4, Mixture of Oligospermia and Azoospermia; 6 mixtures Oligospermia: 343. 6 minutes of average sperm motile time Azoospermia: Oligo.+Azoo. mixture: 348. 1 minutes of average serum motile time In In conclusion, 1. no mutual spermicidal effect but tendency toward enhancement of sperm motile time 2. no sperm agglutinating phenomenon nor sperm immobilizing effect were noted in various semen mixtures 3. posibilities of clinical application such as AID arc considerable and 4. further studies are needed in conjunction with the serum autoimmune mechanism.