Cisplatin-induced azoospermia and testicular damage ameliorated by adipose-derived mesenchymal stem cells

BACKGROUND: The testes are highly susceptible to the adverse effects of chemotherapy and radiation at all stages of life. Exposure to these threats mainly occurs during cancer treatment and as an occupational hazard in radiation centers. The present study investigated the regenerative ability of adipose-derived mesenchymal stem cells (ADMSCs) against the adverse effects of cisplatin on the structure and function of the testes. METHODS: New Zealand white rabbits (N = 15) were divided into three groups of five: a negative control group (no treatment), a cisplatin group (single dose of cisplatin into each testis followed three days later by a PBS injection), and a cisplatin + ADMSCs group (cisplatin injection followed three days later by an ADMSC injection). On day 45 post-treatment, serum testosterone levels were evaluated, and the testes and epididymis were collected for histology, oxidative stress examination, and epididymal sperm analysis. RESULTS: Cisplatin caused damage to the testicular tissue and decreased serum testosterone levels, epididymal sperm counts, and oxidants. An antioxidant imbalance was detected due to increasing malondialdehyde (MDA) and reduced glutathione (GSH) levels in testicular tissue. The ADMSC-treated group displayed a moderate epididymal sperm count, adequate antioxidant protection, suitable hormone levels, and enhanced testicular tissue morphology. CONCLUSIONS: ADMSCs treatment repaired damaged testicular tissue, enhanced biochemical parameters, and modified pathological changes caused by cisplatin.

Classification of the sperm chromatin compaction alterations in bulls (Bos taurus) and its correlation with the efficiency of the in vitro production of embryos

This paper proposes to classify the sperm chromatin compaction alterations in bulls, according to the affected area location and its objective is evaluating the correlation of the intensity, the heterogeneity and these kinds of chromatin decompaction with the rates of cleavage and the formation of blastocysts of in vitro production of embryos (IVPE). It was used several subfertile animals sperm samples, which were evaluated using the toluidine blue staining and computer image analysis, making possible the categorization of the chromatin decompaction according to their location. The percentages of chromatin decompaction and heterogeneity were also evaluated. IVPEs were done and the rates of cleavage and of blastocysts were correlated with the chromatin characteristics. It made possible the classification of the chromatin decompaction according to the head affected part in at least four types: base decompaction, basal half decompaction, central axis decompaction, total decompaction. Based on the correlation, it can be implicated that each type of classification has different influences on the bull fertility. It made possible understanding that sperms amount with 5% or more of chromatin decompaction intensity interferes in the bull fertility and this condition can be featured as an uncompensable defect, while the heterogeneity of chromatin is not an important factor in the IVPE results.

Evaluation of stallion sperm motility with ImageJ using a cell phone camera and a light microscope / [Avaliação da motilidade espermática de garanhões com ImageJ usando-se câmera de celular e microscópio óptico]

This study aimed to determine the accuracy of assessing stallion sperm motility using a light microscope, a cell phone camera, and a free computer-assisted semen analysis (FCASA) package for ImageJ. The total motility of frozen (n=22) and cooled (n=48) equine semen was determined by FCASA and compared to the results of subjective visual analysis (SVA) by two technicians. Frozen samples were also evaluated by a commercial computer-assisted semen analysis (CCASA) system. The Friedman test revealed no significant differences (P>0.05) between cooled samples analyzed by FCASA (38.0) and SVA (technician 1, 40.0; technician 2, 40.0), nor between frozen samples analyzed by FCASA (23.36 ± 15.9), SVA (25.5 ± 18.8 and 25.8 ± 18.5), and CCASA (25.2 ± 18.3). However, mean FCASA results were underestimated by 7.2% compared with CCASA. The correlation between FCASA and CCASA was significant and strong (P<0.0001, r=0.95). Chi-squared tests indicated that FCASA provided similar results (P=0.14) to the reference method (CCASA), but SVA had lower accuracy (P=0.04). ImageJ analysis of cell phone videos captured under a light microscope can be used for estimation of stallion sperm motility with comparable accuracy to commercial systems.(AU)

O objetivo deste estudo foi testar as configurações necessárias para avaliar a motilidade espermática total de garanhões, mediante o uso de ImageJ, microscópio óptico e câmera de celular. Os valores de motilidade total das amostras de sêmen equino congeladas (22) e refrigeradas (48) foram comparados por análise visual (SVA) e pelo plugin do ImageJ (CASAF). Amostras congeladas também foram comparadas por um CASA comercial (CCASA). O teste de Friedman não resultou em diferença estatística (P>0,05) entre as 48 amostras analisadas com CASAF (38,0) e SVA de dois avaliadores (40,0 e 40,0). A comparação das 22 amostras congeladas entre CASAF (23,36±15,9), SVA (25,5±18,8 e 25,8±18,5) e CCASA (25,2±18,3) também não resultou em diferença estatística, sendo que a média dos resultados obtidos com CASAF subestimou a obtida com o CCASA em 7,2%. A correlação entre CASAF e CCASA foi significativamente elevada (r=0,95, P<0,0001). O teste de qui-quadrado resultou em proporção de acertos semelhantes entre o CASAF e o CCASA (P=0,14), enquanto SVA resultou em proporção diferente (P=0,04), indicando menor acurácia. O uso de microscópio óptico e câmera de celular foi útil para obter vídeos de sêmen de garanhões a serem analisados com ImageJ, proporcionando resultados de motilidade total equiparáveis a sistemas comerciais.(AU)

Padronização do Sperm Class Analyzer® (SCA®) para avaliação de espermatozoides equinos criopreservados / [Standardization of the Sperm Class Analyzer® (SCA®) for the evaluation of cryopreserved equine sperm]

Objetivou-se, no primeiro experimento, avaliar o efeito da velocidade de captura de imagens de 25Hz, 30Hz e 50Hz na cinética dos espermatozoides equinos criopreservados. Todas as velocidades mostraram-se adequadas para capturar o movimento espermático (P>0,05). No segundo experimento, objetivou-se avaliar o efeito da deposição de sêmen em lâmina sob lamínula, Leja®10 e 20, na cinética espermática. O uso de lâmina e lamínula foi superior às lejas para manter a LIN e o WOB (P<0,05). No terceiro experimento, objetivou-se avaliar o efeito das concentrações de 25, 50 e 100x106 na cinética espermática. As concentrações de 25 e 50 x106 foram superiores a 100x106 para preservar a LIN, a STR e a BCF e não afetar negativamente a motilidade (P<0,05). No quarto experimento, objetivou-se avaliar o efeito dos diluidores BotuCrio®, BotuSêmen®, TALP sperm e da solução fisiológica na cinética espermática. O BotuCrio® foi superior a todos os diluidores em preservar a BCF e os hiperativos (P<0,05). Conclui-se que o emprego da velocidade de captura entre 25 e 50Hz, a deposição do sêmen entre lâmina e lamínula e a rediluição em diluidor de congelação para atingir 25 a 50x106 de espermatozoides/mL são ideais para o SCA® avaliar, de forma fidedigna, o sêmen equino criopreservado.(AU)

The objective of the first experiment was to evaluate the effect of 25, 30 and 50Hz frame acquisition rate on equine cryopreserved sperm. All frame acquisition rates tested were adequate to capture the sperm movement (P>0.05). The aim of the second experiment was to evaluate the effect of chambers, slide-coverslip, Leja®10 and 20 on sperm movement. The use of slide-coverslip was superior to maintain LIN and WOB (P<0.05). The aim of the third experiment was to evaluate the effect of 25, 50 and 100x106 sperm/mL concentration on sperm movement. Concentrations of 25 and 50x106 sperm/mL were greater than 100x106 to preserve LIN, STR and BCF and did not adversely affect motility (P<0.05). The aim of the fourth experiment was to evaluate the effect of BotuCrio®, BotuSêmen®, TALP sperm and physiological solution on sperm movement. BotuCrio® was superior among other extenders in preserving BCF and hyperactive (P<0.05). It is concluded that the use of the frame acquisition rate between 25 and 50 Hz; the deposition of semen between slide and coverslip and new dilution in the freezing extender to 25-50x106 of sperm/mL is ideal to reliably evaluate cryopreserved equine semen by SCA®.(AU)

Internal quality control products for computer-assisted sperm analysis: Research and application / 中华男科学杂志

Objective@#To investigate the application of the self-made semen quality control (QC) product in internal QC of computer-assisted sperm analysis (CASA).@*METHODS@#CASA was calibrated with high- and low-concentration commercially available semen QC product and meanwhile 15 samples of self-made mixed semen QC product were placed in 75 cryotubes containing liquid nitrogen, followed by CASA of the concentration, motility, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), wobble (WOB) and straightness (STR) of the sperm using standard procedures and 50 days of continuous monitoring. The Makler counting plate was used to measure the concentration and motility of the self-made sperm.@*RESULTS@#The coefficients of variation (CV) of the commercially available semen QC product at high and low concentrations were 6.18% and 7.85%, respectively. CASA showed that the concentration of the self-made QC product was (25.97 ± 1.41) ×10⁶/ml, with a CV of 5.42%, and the sperm motility, VCL, VSL, VAP, LIN, WOB and STR were (22.15 ± 1.75)% (CV = 7.9%), (59.18 ± 2.05) μm/s (CV = 3.46%), (26.79 ± 1.2) μm/s (CV = 4.48%), (34.98 ± 1.4) μm/s (CV = 4.01%), 46.81 ± 1.55 (CV = 3.3%), 60.52 ± 1.3 (CV = 2.15%) and 76.46 ± 1.98 (CV = 2.59%), respectively. The concentration and motility of the self-made sperm detected with the Makler counting plate were (34.39 ± 2.37) ×10⁶/ml (CV = 6.89%) and (38.04 ± 1.69)% (CV = 4.44%), respectively. Levey-Jennings QC charts were plotted for the indicators using the means and standard deviation.@*CONCLUSIONS@#The self-made internal QC product by liquid nitrogen cryopreservation is feasible and effective for monitoring the accuracy and precision of CASA-derived sperm concentration and motion parameters, and it has a smaller CV than the commercially available QC product in measuring sperm concentration.

Diferentes protocolos de congelação de sêmen canino para melhoria de sua cinética e viabilidade / Different canine semen freesing protocols to improve kinetics and viability / Diferentes protocolos de congelación de semen canino para mejoría de su cinética y viabilidad

A maioria dos protocolos utilizados para a criopreservação de sêmen canino, se baseiam em metodologias descritas para outras espécies. Assim, este estudo tem como objetivo avaliar diferentes protocolos de congelação para sêmen desta espécie doméstica. Para tanto, foram utilizados 3 machos, adultos, da raça Buldogue Campeiro, com idades entre 2 a 5 anos e fertilidade comprovada. Foram realizadas 5 colheitas de sêmen de cada animal, pelo método de manipulação digital do bulbo peniano, priorizando a segunda fração do ejaculado. As amostras colhidas foram divididas em 2 grupos, com concentração de 100 x 106 espermatozoides por mL. No grupo 1, as amostras foram diluídas diretamente em meio de congelação comercial Botudog® (Botupharma Biotecnologia Animal). No grupo 2, as amostras foram centrifugadas a 600 g por 10 minutos e em seguida, o pellet foi ressuspendido em meio de congelação comercial Botudog®. As amostras foram envasadas em palhetas de 0,5 mL com concentração de 50 x 106 espermatozoides viáveis. Em seguida, as amostras permaneceram por 1 hora em estabilização a 5ºC. Logo após, transferidas para o vapor de nitrogênio durante 10 minutos, e por fim, mergulhadas em nitrogênio e armazenadas em botijão criogênico. As palhetas foram descongeladas a 46ºC por 15 segundos. Foram avaliados os parâmetros de cinética espermática e integridade de membrana plasmática e acrossomal (IMPA, %). Verificou-se que os parâmetros de motilidade total (%), velocidade linear progressiva (VSL; µm/s), velocidade curvilínea (VCL; µm/s), linearidade (%), percentagem de espermatozoides rápidos (%) e integridade de membrana plasmática e acrossomal avaliados por citometria de fluxo foram superiores no grupo 1, em que as amostras não foram centrifugadas. Estes dados demonstram que, o protocolo para congelação de sêmen canino, utilizando o diluente Botudog®, não preconiza a centrifugação do ejaculado, previamente a congelação.(AU)

Most protocols used for canine semen cryopreservation are based on methodologies described for other species. Thus, this study aims at evaluating different freezing protocols for semen of this domestic species. To this end, 3 adult Bulldog Campeiro males aged 2 to 5 years and proven fertility were used. Five semen samples were collected from each animal using the digital manipulation of the penile bulb method, prioritizing the second fraction of the ejaculate. The collected samples were divided into 2 groups, with a concentration of 100 x 106 sperm per mL. In group 1, the samples were diluted directly into Botudog® commercial freezing medium (Botupharma Animal Biotechnology). In group 2, the samples were centrifuged at 600 g for 10 minutes and then the pellet was resuspended in commercial Botudog® freezing medium. The samples were packed in 0.5 mL straws with a concentration of 50 x 106 viable sperm. Then, the samples remained for 1 hour in stabilization at 5ºC. Afterwards, they were transferred to nitrogen vapor for 10 minutes, and finally, dipped in nitrogen and stored in cryogenic cylinder. The straws were thawed at 46ºC for 15 seconds. Parameters for spermatic kinetics, and plasma and acrosomal membrane integrity (IMPA, %) were evaluated. Total motility (%), progressive linear velocity (VSL; µm/s), curvilinear velocity (VCL; µm/s), linearity (%), percentage of rapid sperm (%) and membrane integrity were found. Plasma and acrosomal samples evaluated by flow cytometry were higher in group 1, where samples were not centrifuged. These data demonstrate that the protocol for canine semen freezing using Botudog® diluent does not recommend centrifugation of the ejaculate prior to freezing.(AU)

La mayoría de los protocolos utilizados para la criopreservación de semen canino se basan en metodologías descritas para otras especies. Por lo tanto, esta investigación tiene como objetivo evaluar diferentes protocolos de congelación para el semen de esta especie doméstica. Con este fin, se utilizaron 3 machos, adultos, de la raza Bulldog Campero, con edades entre 2 a 5 años y fertilidad comprobada. Se realizaron cinco recolecciones de semen de cada animal mediante el método de manipulación digital del bulbo del pene, priorizando la segunda fracción de la eyaculación. Las muestras recolectadas se dividieron en 2 grupos, con una concentración de 100 x 106 espermatozoides por mL. En el grupo 1, las muestras se diluyeron directamente en medio de congelación comercial Botudog® (Botupharma Animal Biotechnology). En el grupo 2, las muestras fueron centrifugadas a 600 g durante 10 minutos y luego el pellet fue resuspendido en medio de congelación comercial Botudog®. Las muestras se envasaron en paletas de 0,5 mL con una concentración de 50 x 106 espermatozoides viables. Luego, las muestras permanecieron durante 1 hora en estabilización a 5ºC. Posteriormente, se transfirieron al vapor de nitrógeno durante 10 minutos y, finalmente, se sumergieron en nitrógeno y se almacenaron en un cilindro criogénico. Las paletas se descongelaron a 46ºC durante 15 segundos. Se han evaluado los parámetros de la cinética espermática y la integridad de la membrana plasmática acrosomal (IMPA, %). Se verificó que los parámetros de motilidad total (%), velocidad lineal progresiva (VSL; µm/s), velocidad curvilínea (VCL; µm/s), linealidad (%), porcentaje de espermatozoides rápidos (%) e integridad de la membrana plasmática y acrosomal evaluadas por citometría de flujo, fueron mayores en el grupo 1, donde las muestras no fueron centrifugadas. Estos datos demuestran que, el protocolo para la congelación de semen canino, usando el diluyente Botudog®, no preconiza la centrifugación del eyaculado, previamente a la congelación.(AU)

Hamilton-Thorne Integrated Visual Optical System Ⅰ (IVOSⅠ) versus IVOS Ⅱ for human sperm analysis / 中华男科学杂志

Objective@#To compare the computer-assisted sperm analysis (CASA) systems Hamilton-Thorne Integrated Visual Optical System Ⅰ (IVOSⅠ) and IVOS Ⅱ after verifying the performance of the latter so as to ensure the accuracy of the results of analysis.@*METHODS@#Based on the criteria established in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen (WHO 5th Ed), we compared the main semen parameters obtained from IVOSⅠ with those generated by IVOS Ⅱ, and examined the consistency between the results of the two sperm analyzers.@*RESULTS@#The linear relationship of the outlier test, bias estimation and scatter plot and the results of the outlier test of the two systems all met the requirements of comparison analysis and showed an obvious correlativity. The application scope of the results obtained from the apparatus indicated a reasonable value range, with r = 0.988 for sperm concentration, r = 0.975 for sperm progressive motility (PR), and r = 0.981 for total sperm motility. Evaluation of the acceptability of the predicted bias showed that the allowable total error (TEa) to be 6.67% with sperm concentration at 12 × 106 /ml and 2.34% with PR < 31%, their upper limit of the allowable error < 1/2. The results of IVOS Ⅱ conformed to the requirements of the WHO 5th Ed.@*CONCLUSIONS@#The main parameters derived from IVOSⅠ and IVOS Ⅱ are comparable and consistent, indicating that both can be used for the examination of semen samples.

Efeito da adição de antioxidantes sobre a viabilidade do sêmen bovino / Addition of antioxidants on the feasibility of bovine semen / Efecto de la adición de antioxidantes bajo la viabilidad del semen bovino

Os antioxidantes atuam no combate aos efeitos nocivos provocados pelos radicais livres, proporcionando melhor qualidade do sêmen fresco e criopreservado. As mudanças de temperatura provocadas durante os processos de congelação e descongelação são responsáveis pelos danos aos espermatozoides, alterando sua viabilidade e fertilidade. O estudo dos antioxidantes, os quais constituem a primeira linha de combate às espécies reativas de oxigênio (EROs), para a melhora da qualidade do sêmen criopreservado, torna-se de fundamental importância, para desenvolvimento de protocolos padrões e aplicáveis, uma vez que existem muitas controvérsias em relação ao tipo de molécula antioxidante e às doses, o momento adequado para ser acrescentado e possíveis alterações destes aos meios de criopreservação. Assim, este trabalho tem como objetivo estudar o efeito dos antioxidantes: catalase, α-tocoferol e piruvato de sódio adicionados em meio diluidor comercial para melhorar a viabilidade do sêmen bovino criopreservado. Inicialmente foram utilizados três touros, da raça Brahman, selecionados de acordo com o escore de condição corporal e exame andrológico. Oito ejaculados foram colhidos por eletroejaculação e imediatamente avaliados a motilidade e o vigor espermático. Em seguida, foram diluídos conforme os grupos experimentais em meio Botubov® (Botupharma Biotecnologia Animal) com adição do antioxidante piruvato de sódio nas concentrações de 1,5 µM, 3 µM e 5 µM e em seguida, as amostras foram congeladas. Numa segunda etapa, serão utilizados outros antioxidantes e concentrações, conforme descrito: 1) Diluidor BotuBov®; 2) Diluidor BotuBov® contendo 7% de crioprotetor e catalase nas concentrações de 20, 80 e 200 UI; 3) Diluidor BotuBov® contendo 7% de crioprotetor e α-tocoferol nas concentrações de 50 µM, 100 µM e 150 µM; 4) Diluidor BotuBov® contendo 7% de crioprotetor e piruvato de sódio nas concentrações de 1,5 µM; 3,5 µM e 5 µM. Após a descongelação do sêmen com a utilização do piruvato, as amostras foram analisadas quanto à motilidade total e progressiva e vigor espermático. Verificou-se que a adição de piruvato de sódio na concentração de 1,5 µM proporcionou uma melhora nos parâmetros de motilidade total e vigor espermático, demonstrando os benefícios de seu uso.(AU)

Antioxidants act in fighting the harmful effects caused by free radicals, providing better quality of both fresh and cryopreserved semen. Changes in temperature caused during the freezing and thawing processes are responsible for damages to the sperm, changing their viability and fertility. The study of antioxidants, which constitute the first line of combat against reactive oxygen species (ROS) to improve the quality of cryopreserved semen, is of utmost importance for the development of standard and applicable protocols, since there are many controversies regarding the type and the doses of antioxidant molecules, the timing for its addition and likely changes to the cryopreservation media. This paper aims at studying the effects of catalase, α-tocopherol, and sodium pyruvate when added to commercial diluent medium to improve the viability of cryopreserved bovine semen. Initially, three Brahman bulls were selected according to the body condition score and andrological examination. Eight ejaculates were collected by electroejaculation, with motility and spermatic vigor being immediately assessed. Subsequently, the samples were diluted according to the experimental groups in Botubov® (Botupharma Animal Biotechnology) medium with the addition of sodium pyruvate at concentrations of 1.5 µM, 3 µM and 5 µM and were subsequently frozen. In a second step, other antioxidants and concentrations will be used, as follows: 1) BotuBov® Diluent; 2) BotuBov® diluent containing 7% cryoprotectant and catalase at concentrations of 20, 80 and 200 IU; 3) BotuBov® diluent containing 7% cryoprotectant and α-tocopherol at concentrations of 50 µM, 100 µM and 150 µM; 4) BotuBov® diluent containing 7% cryoprotectant and sodium pyruvate at concentrations of 1.5 µM; 3.5 µM and 5 µM. After thawing the semen with pyruvate, the samples were analyzed for total and progressive motility and spermatic vigor. It was found that the addition of 1.5 µM sodium pyruvate provided an improvement in total motility and sperm vigor parameters, demonstrating the benefits of its use.(AU)

Los antioxidantes actúan en el combate a los efectos nocivos provocados por los radicales libres, proporcionando mejor calidad del semen fresco y criopreservado. Los cambios de temperatura provocados durante los procesos de congelación y descongelación son responsables por los daños a los espermatozoides, alterando su viabilidad y fertilidad. El estudio de los antioxidantes, que constituyen la primera línea de combate a las especies reactivas de oxígeno (ERO), para la mejora de la calidad del semen criopreservado, se hace de fundamental importancia para el desarrollo de protocolos estándares y aplicables, ya que existen muchas controversias en relación al tipo de molécula antioxidante y a las dosis, el momento adecuado para ser añadido y posibles alteraciones de éstos a los medios de criopreservación. Así, este estudio ha tenido como objetivo estudiar el efecto de los antioxidantes: catalasa, α-tocoferol y piruvato de sodio añadidos en medio diluidor comercial para mejorar la viabilidad del semen bovino criopreservado. Inicialmente se utilizaron tres toros, de la raza Brahman, seleccionados de acuerdo con la puntuación de condición corporal y examen andrológico. Ocho eyaculados fueron recogidos por electroejaculación e inmediatamente evaluados la motilidad y el vigor espermático. A continuación, se diluyeron de acuerdo con los grupos experimentales en medio Botubov (Botupharma Biotecnología Animal) con adición de antioxidante piruvato de sodio a concentraciones de 1,5 µM, 3 µM y 5 µM, y luego las muestras se congelaron. En una segunda etapa, se utilizarán otros antioxidantes y concentraciones, según se describe: 1) Diluidor BotuBov®; 2) Diluidor BotuBov® conteniendo 7% de crioprotector y catalasa en las concentraciones de 20, 80 y 200 UI; 3) Diluidor BotuBov® que contiene 7% de crioprotector y α-tocoferol en las concentraciones de 50 µM, 100 µM y 150 µM; 4) Diluidor BotuBov® conteniendo 7% de crioprotector y piruvato de sodio en las concentraciones de 1,5 µM; 3,5 µM y 5 µM. Después de la descongelación del semen con la utilización del piruvato, las muestras fueron analizadas en cuanto a la motilidad total y progresiva y vigor espermático. Se comprobó que la adición de piruvato de sodio en la concentración de 1,5 µM proporcionó una mejora en los parámetros de motilidad total y vigor espermático, demostrando los beneficios de su uso.(AU)

Sperm agglutinating activity of Saccharomyces cerevisiae and Candida albicans as a potential causative factor of infertility in mice (Mus musculus)

@#<p style="text-align: justify;"><strong>BACKGROUND AND OBJECTIVE:</strong> Vaginal yeast infections in women are usually caused by Candida albicans and, to a lesser extent, by Saccharomyces cerevisiae. Studies on C. albicans have shown that it can cause sperm agglutination which can lead to lowered fertility. This study was conducted to compare the effect of S. cerevisiae and C. albicans on the fertility of ICR mouse (Mus musculus) through sperm agglutination.</p><p style="text-align: justify;"><strong>METHODOLOGY:</strong> Sperm agglutinating activity was examined by mixing different concentrations of S. cerevisiae (10, 10°, and 10 CFU/mL) and C. albicans (10", 10°, and 10 CFU/mL) separately with semen from male mice of ICR strain. Determination of the effect of S. cerevisiae and C. albicans on the fertility outcome of female mice was done by intravaginal inoculation of 20 uL of 104, 106, and 108 CFU/ml of the two yeast organisms and later allowed to mate.</p><p style="text-align: justify;"><strong>RESULTS AND CONCLUSION:</strong> The study showed a statistically significantly higher percent sperm agglutination by S. cerevisiae than C. albicans at 10* CFU/ml but no difference was observed at 10° and 10 CFU/ml. No significant difference was observed in the number of sperm per agglutinate between the two yeast species at a=0.05. The concentration that exhibited the highest percentage of agglutinated sperm is 10° CFU/mL for both yeast. The most frequent type of agglutination observed in S. cerevisiae is the mixed type, while head-to-head type is most frequent in C. albicans. Both yeasts were able to cause a decline in the number of births in mice starting at 10 CFU/ml. While sperm agglutination could be one of the reasons for the infertility observed in mice, there may be other processes, mechanisms, and/or activities that could contribute to such an outcome.</p>

Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs / Teste de ligação espermática à membrana perivitelínica da gema de ovo de galinha como avaliação funcional do sêmen de cães

During fertilization, spermatozoa interact with the zona pellucida (ZP) through the binding between the acrosome and proteins 2 and 3 (ZP2 and ZP3). The perivitelline membrane of chicken egg yolk is homologous to the mammalian ZP3, which allows the binding of sperm of several species. The aim of this study was to standardize and evaluate the efficiency of sperm-binding to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation. For this purpose, nine post-thaw sperm samples were used, which were divided into two aliquots: the first was kept in water bath at 37ºC (live sample) and the second was submitted to cold shock to induce cellular damage (dead sample). The two aliquots were mixed on five proportions, corresponding to 0%, 25%, 50%, 75%, and 100% of viable cells, and the binding test was performed by analyzing the number of spermatozoa bonded to the perivitelline membrane by means of computerized assessment of sperm motility (CASA) or conventional microscopy. Additionally, samples were submitted to sperm motility analysis, evaluation of plasmatic and acrosomal membrane integrity, and sperm mitochondrial activity. The sperm-binding test to the perivitelline membrane of chicken egg yolk was considered a feasible sperm analysis test for both fertilizing capacity and overall sperm attributes evaluation, mainly when the analysis is performed by a conventional microscope, which expands its practicality to the majority of canine reproduction laboratories.(AU)

Durante a fecundação, os espermatozoides interagem com a zona pelúcida (ZP) por meio da ligação entre o acrossomo e as proteínas 2 e 3 (ZP2 e ZP3). A membrana perivitelínica da gema de ovo de galinhas é homóloga à ZP3 de mamíferos, possibilitando a ligação espermática de diversas espécies. Este trabalho padronizou e avaliou a eficiência do teste de ligação espermática à membrana perivitelínica da gema de ovo de galinhas como avaliação funcional do sêmen de cães. Para tal, foram utilizadas nove amostras seminais previamente criopreservadas. Cada amostra foi dividida em duas alíquotas: a primeira foi mantida em banho-maria à 37ºC (vivos) e a segunda submetida a choque térmico com o intuito de induzir dano celular (mortos). As duas alíquotas foram misturadas, correspondendo a 0, 25, 50, 75 e 100% de células viáveis. As amostras foram avaliadas quanto ao número de espermatozoides ligados à membrana perivitelínica por meio da análise computadorizada da motilidade (CASA) ou microscopia convencional. Ademais, as amostras foram avaliadas quanto à motilidade espermática, integridade das membranas acrossomal e plasmática e atividade mitocondrial espermática. O teste de ligação espermática à membrana perivitelínica de ovos de galinha foi considerado um teste de análise seminal exequível tanto para avaliar a capacidade fecundante dos espermatozoides como atributos seminais gerais, especialmente quando realizado em microscopia convencional, expandindo sua praticidade para a maioria dos laboratórios de análise de sêmen canino.(AU)

Transcutaneous electrical acupoint stimulation for asthenozoospermia / 中华男科学杂志

Objective@#To study the effect of transcutaneous electrical acupoint stimulation (TEAS) in the treatment of asthenozoospermia.@*METHODS@#We randomly divided 72 asthenozoospermia patients into a 2 Hz TEAS (n = 29), a 100 Hz TEAS (n = 20), and a blank control group (n = 23), those in the former two groups treated by 30 minutes of TEAS at 2 Hz and 100 Hz respectively, applied to the acupoints of bilateral Shenshu, left Zusanli, and Guanyuan, once a day for 60 days, while those in the blank control group left untreated. Using computer-assisted sperm analysis (CASA), we examined sperm concentration and motility as well as the percentages of grade a and grade a+b sperm in different groups of the patients.@*RESULTS@#Compared with the baseline, 2 Hz TEAS significantly increased sperm motility (［12.76 ± 1.39］ vs ［18.89 ± 2.46］%, P<0.05) and the percentage of grade a+b sperm ( ［10.68 ± 1.22］ vs ［16.32 ± 2.10］%, P<0.05) in the asthenozoospermic patients, while 100 Hz TEAS improved not only sperm motility (［12.32 ± 2.21］ vs ［23.81 ± 3.42］%, P<0.01) and the percentage of grade a+b sperm (［10.45 ± 1.98］ vs ［20.25 ± 2.82 ］%, P<0.01), but also the percentage of grade a sperm (［6.44 ± 1.16］ vs ［13.31 ± 2.30］%, P<0.05). Moreover, in comparison with the blank control group, 2 Hz TEAS also remarkably increased sperm motility (［9.57 ± 1.60］ vs ［18.89 ± 2.46］%, P<0.05) and the percentage of grade a+b sperm (［7.81 ± 1.31］ vs ［16.32 ± 2.10］%, P<0.05) in the asthenozoosperma patients, while 100 Hz TEAS improved not only sperm motility (［9.57 ± 1.60］ vs ［23.81 ± 3.42］%, P<0.01) and the percentage of grade a+b sperm (［7.81 ± 1.31］ vs ［20.25 ± 2.82］%, P<0.01) but also the percentage of grade a sperm (［4.87 ± 1.01］ vs ［13.31 ± 2.30］%, P<0.01). Meanwhile, the rate of clinical effectiveness was significantly higher in the 100 Hz TEASthan in the blank control group either in intention-to-treat (ITT) analysis (100% vs 18.18%) orper-protocol (PP) analysis (90% vs 0%), and so was it than in the 2 Hz TEAS group based on the data of ITT (100% vs 33.33%).@*CONCLUSIONS@#Both 2 Hz and 100 Hz TEAS are effective for the treatment of asthenozoospermia by improving sperm motility and vitality.

Clinical Observation of Acupuncture for Idiopathic Oligoasthenozoospermia / 上海针灸杂志

Objective To observe the clinical efficacy of acupuncture in treating male idiopathic oligoasthenozoospermia. Method A hundred patients with oligoasthenozoospermia were recruited and randomized into a treatment group and a control group, 50 cases in each group. The treatment group was intervened by acupuncture, while the control group was by placebo acupuncture at non-meridian points. The clinical efficacies and changes of the relevant parameters of sperms after treatment were observed. Result The clinical efficacy of the treatment group was significantly better than that of the control group (P0.05); after intervention, the improvements of the relevant parameters of sperms in the treatment group were more significant than that in the control group (P<0.05). Conclusion Acupuncture can effectively improve the sperm density, sperm motility, and sperm viability, and enhance the recovery rate in treating infertility, and it’s easy-to-operate, without significant adverse reactions, safe, and reliable.

Morphometric and kinematic sperm subpopulations in split ejaculates of normozoospermic men / 亚洲男科学杂志(英文版)

This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis (CASA-Mot), and of sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations (slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%]), and three morphometric subpopulations (large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]). The distribution of kinematic sperm subpopulations was different among ejaculate fractions (P < 0.001), with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions (P < 0.001), with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.

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.

The role of bacterial infections on male infertility in Al-Anbar province of Iraq.

Background: Bacterial infections on male infertility has always been in the field of debate due to scarce analysis tools to examine seminal fluid specimens as a result of which these infectious processes leads to deterioration of spermatogenesis, impairment of sperm function and/or obstruction of the seminal tract. Aims & Objective: In the current study we investigated the role of bacterial infections in male factor infertility in Al-Anbar Province, West of Iraq through detection of abnormal sperms and other factor pertains to male infertility. Material and Methods: Seminal fluid from six hundred volunteer males was investigated for infertility by the detection of abnormal sperms using the WLJY-9000 TYPE WEILI Color Sperm Analysis System and the Neubauer counting chamber. Results: From the six hundreds patients investigated for infertility, it was found that 408 (68%) patients had a positive culture for pathogenic bacteria, of different species. The results indicate that 32.0% had sperm density less than twenty million per millilitre. The oligospermic were 23.0%, severe oligospermic 0.17% and Azoospermia 8.83%. Asthenospermia was reported to be 76.33% and Teratospermia 86.16% respectively. Conclusion: Seminal fluid infection increases with decreasing sperm density, motility and morphology. The prevalence of abnormal sperm indices and bacterial infection is high with Klebsiella spp. infection. Hence, treatment measures should be taken properly in the management of male factor infertility.

O impacto da criopreservação na qualidade seminal / The impact of the cryopreservation on seminal quality

Introdução: as indicações iniciais para o uso da técnica de fertilização in vitro contemplavam boa parte dos diagnósticos de infertilidade feminina. Com o passar do tempo, tornou-se necessário o desenvolvimento de outras tecnologias que tratassem também os casos de infertilidade masculina. Dentre elas, destaca-se a criopreservação de gametas. Apesar das vantagens do uso da criopreservação, sabe-se que alguns processos de congelamento podem afetar o potencial de fertilidade em muitos aspectos. Objetivo: por meio de uma revisão, verificar o impacto da criopreservação na qualidade do sêmen por meio da observação da taxa de gestação, taxa de aborto e das características seminais (morfologia, motilidade, concentração, fragmentação do DNA) no sêmen congelado e o no fresco, colhidos diretamente do epidídimo. Método: foi feita uma estratégia de busca nas bases de dados Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs), Medical Literature Analysis and Retrieval System Online (Medline) e Biblioteca Cochrane por meio das seguintes palavras-chaves: sêmen, criopreservation, frozen sêmen, thawed sêmen e sêmen quality. As pesquisas foram feitas tanto nos artigos e nas revisões disponíveis em full text como nos resumos. Resultados: de maneira geral, o que se observou foi que em alguns casos a criopreservação pode prejudicar a morfologia seminal. Porém, a quantidade de nascimentos e de abortos não varia significativamente quando comparado sêmen fresco com congelado obtidos diretamente do epidídimo. Conclusões: por prolongar a fertilidade de muitos pacientes e ajudá-los na realização do sonho da paternidade, na maioria dos casos, a criopreservação é uma boa técnica que deve ser usada quando necessário.

Introduction: the initial indications to the use of in vitro fertilization technologie attented almost all the female infertility cases. Over the time, the development of others technologies that could treat the male infertility cases too became necessary. Among the technologies, itcan be standed out the gametes cryopreservation. Despite the cryopreservation advantages,it is known that some freezing processes can affect fertility potential in many ways. Objective: verify the impact of cryopreservation on semen quality by observing the pregnancy rate, abortion rate and seminal characteristics (morphology, motility, concentration andDNA) fragmentation between frozen semen and fresh semen harvested directly from the epididymis, through a review. Method: for this, it was done a research in the databases Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs), Medical Literature Analysis and Retrieval SystemOnline (Medline) and Cochrane using the following keywords semen cryopreservation, frozen semen, thawed semen and semen quality. The surveys were made both in articles andreviews available in full text as in the summaries. Results: in general, the seminal cryopreservation may damage the morphology of semen. However, the number of births and abortions does not vary significantly when comparedfrozen semen with fresh semen obtained directly from the epididymis. Conclusions: thus, by extending the fertility of many patients, helping them in achieving the dream of paternity in most cases, cryopreservation is a good technique that can be used when necessary.

Assessment of chlorpyrifos toxicity on certain organs in rat, Rattus norvegicus.

Chlorpyrifos, an organophosphate insecticide of phosphorothioate group was orally administered to male rats at the doses of 3, 6 and 9 mg kg-1d-1 for 90 days. Animals exposed to high dose (9 mg kg-1d-1) showed signs of toxicity including piloerection, diarrhoea, nose and eye bleeding, reduced body weight and death of animals. Organ weight ratio of different vital organs did not show any change except increase in adrenal weight and decrease in the weight of testes in animals of high dose (9 mg kg-1d-1). A dose dependent inhibition of acetylcholinesterase (AChE) activity in RBC (22-60%) and brain (7-52%) was observed. Microscopic examination of different tissues of male rats showed minor histopathological changes in brain, liver, testis, epididymis and adrenal. The activity of testicular enzymes SDH, G-6-PDH and testicular content of sialic acid and cholesterol were found increased in animals of high dose (9 mg kg-1d-1). There was decrease in RBC counts and levels of hemoglobin (Hb) and hematocrit (HCT) with increase in WBC counts. While, total protein was decreased significantly at all the dose levels in testes and epididymis, glucose level showed a significant decrease at high dose. A dose dependent increase was observed in the level of serum triglycerides. There was no change in sperm motility and sperm morphology at any dose level except a decrease in sperm counts (114.1x 106 g-1 in high dose for group against 158.9 x 106 g-1 controls). It is suggested that chlorpyrifos at 9mg/kg/d dose for 90 days has caused toxicological changes along with mild testicular and spermatotoxic effects in male rats.

Correlation of Post Swim-Up Acrosome Index with in-vitro Fertilization Outcomes

PURPOSE: A prospective study was planned to determine the relationship between post swim-up acrosome index (AI) evaluation and fertilization outcomes in an in vitro fertilization (IVF) program. MATERIALS AND METHODS: Infertile couples who have applied to IVF were admitted into this study when the male partner's sperm concentration was > 20x106/mL and motility > 30%. Pre- and post swim-up semen quality parameters including concentration, motility, sperm morphology and AI were evaluated in a prospective, randomized and blinded fashion. The couples were divided prospectively into 2 groups. In group I (25 couples) 50 000 sperm per oocyte were used for insemination considering post swim-up acrosome index, and in group II (25 couples) 50 000 sperm per oocyte were used for insemination without considering post swim-up acrosome index. RESULTS: Pre- and post swim-up AI were 30.8 +/- 3.4 and 17.8 +/- 4.5 in group I, and 31.4 +/- 3.6 and 16.3 +/- 4.7 in group II (p > 0.05) respectively. The significant improvement in morphology and motility after double wash swim-up procedure has been observed. However, double wash swim-up procedure could not eliminate head and especially acrosomal defects which would directly effect fertilization capacity in conventional IVF program. In group I, 85.3% of oocytes were fertilized, with a 48% pregnancy rate; in group II, 71.0% of oocytes were fertilized, with a pregnancy rate of 20%. Fertilization and pregnancy rates were significantly different (p < 0.05) between the two groups. CONCLUSION: We have concluded that it could be useful to consider post swim-up AI of sperm inseminated in conventional IVF cycles, which correlates with high fertilization and pregnancy rates.

Sperm analysis among 120 fertile Filipino males aged 20-40 years

In the Philippines, there is no published data on standard normal values of sperm analysis among fertile Filipino males. This study aims to show the semen parameters of healthy fertile Filipino males aged 20-40 years. Convenience sampling was used. One hundred twenty volunteers were recruited and the semen samples were analyzed for sperm density, motility and morphology. Sperm density ranged from 17.1 to 201.6 million/ml (median 63.3 million/ml). Normal tail morphology range was 55 to 99% (median 85%). Normal tail morphology ranged from 55 to 99% (median 71.5%). These correlate with the standard set by the World Health Organization and is consistent with studies done abroad.