Evaluación del factor masculino mediante espermograma más práctico y económico / Sperm Analysis for Assessing Male Factor More Practical and Economical

Es aceptado que la infertilidad se evalúa cuando una pareja no logra embarazarse después de 12 meses de coitos regulares sin protección, problema que afecta a un alto porcentaje de parejas en el mundo. Generalmente, el diagnóstico y tratamiento de la infertilidad se enfoca solo desde el punto de vista femenino, por la obvia relación con el embarazo; sin embargo, con la evidencia existente el problema debe ser abordado como una alteración de pareja(AU)

It is accepted that infertility is evaluated when a couple does not become pregnant after 12 months of regular unprotected intercourse. This problem affects a large percentage of couples worldwide. Generally, diagnosis and treatment of infertility approaches only the female point of view, for the obvious connection with the pregnancy. However, this problem must be addressed with the actual evidence as an alteration in the couple(AU)

Fragmentación del ADN espermático e infertilidad masculina / Fragmentation of sperm DNA and male infertility

La integridad del ADN de los espermatozoides, es un indicador importante de la fertilidad, se utiliza como una variable adicional para evaluar, junto al espermograma, la calidad de una muestra seminal. En el presente trabajo se describen algunos aspectos de interés relacionados con la fragmentación del ADN espermático, cuya etiología es multifactorial y está relacionada con factores intrínsicos, como el empaquetamiento anormal de la cromatina durante la espermiogénesis, la apoptosis defectuosa antes de la eyaculación y la producción excesiva de especies reactivas del oxígeno; y con factores extrínsecos, como son los cambios en los hábitos de vida, la exposición a agentes tóxicos, así como la edad avanzada. La fragmentación está asociada al deterioro de las variables seminales, además afecta la fertilidad natural y la realizada por tratamientos de reproducción asistida, por lo que la implementación en los laboratorios de Andrología, de la tecnología para detectar si el ADN de los espermatozoides está íntegro o fragmentado, incorpora un nuevo conocimiento en el estudio de los hombres con trastornos de fertilidad, lo que contribuye a mejorar el diagnóstico y pronóstico de la infertilidad masculina. Para realizar este trabajo se revisaron 122 artículos, de los cuales 84 cumplieron con los criterios de calidad esperados. La búsqueda se realizó a través de los buscadores habituales(AU)

Sperm DNA´s integrity is an important indicator of fertility and it is used as an additional variable to evaluate the quality of a seminal sample, together with the spermogram. This paper describes some interesting aspects related to the fragmentation of sperm DNA, whose etiology is multifactorial and is related to intrinsic factors, such as the abnormal packing of chromatin during spermiogenesis, defective apoptosis before ejaculation, and the excessive production of oxygen´s reactive species; and with extrinsic factors, such as changes in life habits, exposure to toxic agents, as well as aging. Fragmentation is associated with the deterioration of seminal variables, it also affects natural fertility and that produced by assisted reproduction treatments, so the implementation in the andrology laboratories of the technology to detect if the DNA of the sperm is intact or fragmented incorporates a new knowledge in the study of men with fertility disorders, which contributes to improve the diagnosis and prognosis of male infertility. To carry out this work, 122 articles were reviewed, of which 84 met the expected quality criteria. The search was made through the usual search engines(AU)

Sensitivity evaluation of the computer-assisted sperm analysis (CASA) in the determination of frozen-thawed bull semen concentration / Avaliação da sensibilidade da técnica computadorizada de análise (CASA) para a determinação da concentração espermática do sêmen bovino congelado

Sperm concentration is traditionally evaluated by counting cells in a hemocytometric Neubauer chamber, often a highly subjective, time-consuming, and laborious technique prevalent in andrology laboratories around the world. However, the Computer-Assisted Semen Analysis (CASA) represents a more consistent method of evaluating sperm concentration that may provide enhancing efficiencies of sperm count. The purpose of this study is to compare the results of these two methods in the analysis of post-thaw concentration of bovine semen. Four hundred and twenty five batches of semen from different bulls were selected, thawed at 37°C for 30 seconds and then homogenized. Aliquots of 40 µL of semen were diluted in 960 µL of distilled water, fixing the rate at 1:25 dilution for analysis in a Neubauer chamber. Conversely, aliquots of 5 µL for each semen dose were submitted to CASA, considered a minimum of five random fields and 2000 sperm count per analysis. The average concentration of sperm cells was 38.96a ± 1.28 in the Neubauer analysis and 35.14b ± 0.82 for the CASA, with the correlation coefficient of 0.87 (P < 0.0001) and reliability of 0.78 (scale ranging from 0 to 1) between the two methods. In conclusion, the results of two techniques for assessing sperm concentration have similar results. However the CASA methodology would yield greater benefit due to precision, consistency, and reduced disposal issues, particularly for large processing laboratories.(AU)

Tradicionalmente, a concentração espermática é avaliada por meio da contagem de células em câmara hemocitométrica de Neubauer, técnica laboriosa adotada na rotina dos laboratórios de andrologia. Uma alternativa para essa contagem é a técnica computadorizada de avaliação espermática (CASA), método que pode aumentar a eficiência e acurácia na determinação da concentração de espermatozoides em uma amostra de sêmen. O presente trabalho relata a avaliação da sensibilidade da técnica CASA para o acesso da concentração de espermatozoides bovinos em pósdescongelação. Foram selecionadas 425 doses de sêmen de reprodutores de diferentes raças, descongeladas a 37°C por 30 segundos e homogeneizadas. Alíquotas de 40 µL de sêmen foram transferidas para tubos cônicos de 1,5 mL previamente preenchidos com 960 µL de água destilada, fixando a taxa de diluição em 1:25 para contagem em câmara de Neubauer. Em contrapartida, alíquotas de 5 µL de cada dose de sêmen foram avaliadas com o emprego do sistema CASA considerando o número mínimo de cinco campos aleatórios e 2 mil espermatozoides por análise. A concentração média de células espermáticas foi de 38,96a ± 1,28 e 35,14b ± 0,82,respectivamente para amostras avaliadas em câmara de Neubauer ou sistema computadorizado, apresentando o coeficiente de correlação de 0,87 (P < 0.0001) e concordância de 0,78 (escala de 0 a 1). Conclui-se que as duas técnicas de avaliação da concentração espermática possuem eficiência similar. No entanto, em virtude da precisão, rapidez e por dispensar a diluição prévia das amostras para a contagem, a CASA é uma alternativa para a contagem de células espermáticas em câmara de Neubauer, sobretudo para grandes centrais de produção de sêmen bovino congelado.(AU)

Características andrológicas e do sêmen de touros do composto Red Norte (Nelore x Tabapuã x Red Angus x Sinepol) / Andrological and semen characteristics of cross-bred Red Norte (Nelore x Tabapuã x Red Angus x Sinepol) young bulls

Avaliaram-se as características andrológicas do sêmen de touros jovens do composto Red Norte (Nelore x Tabapuã x Red Angus x Sinepol), com idade média de 13,9±0,8 meses, com o objetivo de estimar o advento da puberdade e a qualidade do sêmen. Foram avaliados o perímetro escrotal (PE), o peso e as características seminais de 70 tourinhos, classificados em três grupos, de acordo com o PE: GI=27-33cm (n=24), GII=33-35cm (n=24) e GIII=35-43cm (n=22). As médias de peso e a idade de cada grupo (G) foram, respectivamente: GI=411,2±37,4kg e 13,8±1,0 meses, GII=426,9±31,5kg e 14,0±0,7 meses e GIII=438,4±38,3kg e 14,0±0,6 meses. As características seminais para cada grupo foram, volume 4,2±3,1mL, 5,3±2,6mL e 4,5±2,1mL; motilidade 31,3±24,1 por cento, 44,2±23,9 por cento e 43,9±21,5 por cento e vigor 2,8±1,6, 3,5±1,3 e 3,5±1,3, respectivamente. O espermiograma apresentou valores médios de concentração de 130,5±266,2x10(6)/mL, 289,5±390,2x10(6)/mL e 333,9±523,7x10(6)/mL, defeitos totais de 81,4±15,9 por cento, 73,8±15,4 por cento e 67,9±19,0 por cento; defeitos maiores de 87,3±26,2 por cento, 66,8±24,9 por cento e 56,7±17,1 por cento e defeitos menores de 16,6±14,9 por cento, 33,2±24,9 por cento e 43,3±17,1 por cento, respectivamente. Dos setenta animais examinados, sete (10 por cento) foram considerados aptos à reprodução. Os resultados mostraram que a patologia espermática diminuiu em razão do aumento do PE.

Reproductive traits of cross-breed Red Norte (Nelore x Tabapuã x Red Angus x Sinepol) young bulls averaging of 13.9±0.8 month-old were evaluated, in order to determine the puberty onset and semen quality in these animals. Scrotal circumference (SC), body weight (BW), and semen parameters of 70 bulls were measured. Animals were allotted in three groups (G) according to their SC: GI=27-33cm (n=24), GII=33-35cm (n=24), and GIII=35-43cm (n=22). BW and age of each group were, respectively: GI=411.2±37.4kg and 13.8±1.0 month-old, GII=426.9±31.5kg and 14.0±0.7 month-old, and GIII=438.4±38.3kg and 14.0±0.6 month-old. Seminal physical characteristics for same order of groups were: volume 4.2±3.1mL, 5.3±2.6mL, and 4.5±2.1mL; motility 31.3±24.1 percent, 44.2±23.9 percent, and 43.9±21.5 percent; and vigor 2.8±1.6, 3.5±1.3, and 3.5±1.3. The spermiogram presented concentration values of 130.5±266.2x10(6)/mL, 289.5±390.2x10(6)/mL, and 333.9±523.7x10(6)/mL; total defects of 81.4±15.9 percent, 73.8±15.4 percent, and 67.9±19.0 percent; major defects of 87.3±26.2 percent, 66.8±24.9 percent and 56.7±17.1 percent; and minor defects of 16.6±14.9 percent, 33.2±24.9 percent, and 43.3±17.1 percent, for same order of groups. Seven out of 70 bulls were considered satisfactory potential breeders. Results showed that semen pathology progressively decreased when SC increased.

Utilização do espermatócrito para estimar aconcentração espermática no sêmen da piabanha(Brycon insignis) / Use of spermatocrit to estimate sperm concentration of semen from piabanha (Brycon insignis)

O objetivo deste trabalho foi verificar a viabilidade da utilização doespermatócrito para estimar a concentração espermática do sêmen depiabanha. Os exemplares (n=29) de piabanha Brycon insignis foramhipofisados, sendo posteriormente realizada a coleta de sêmen. Oespermatócrito foi determinado utilizando-se micro-centrífuga detubos capilares. A concentração espermática foi verificada por contagemem câmara de Neubauer, após diluição. A concentração espermáticaverificada (média±desvio padrão) foi de 24,38 ± 3,84 x 109espermatozóides / mL, e o espermatócrito foi de 16,14±5,20 %. Aequação de regressão: = 14,01.109 + 0,6428.109.X foi significativa(P<0,01) e apresentou R2 ajustado = 0,75. Deste modo, é possívelestimar a concentração através da determinação do espermatócrito.

Avaliação do espermograma de leões africanos (Panthera leo, Linnaeus, 1758), mantidos na Fundação Parque Zoológico de São Paulo / Spermogram of african lions (Panthera leo, Linnaeus, 1758) housed at Fundação Parque Zoológico de São Paulo

O presente estudo teve por objetivo avaliar o espermograma de um grupo de leões africanos mantidos em cativeiro na Fundação Parque Zoológico de São Paulo, como um aspecto a ser considerado na avaliação do potencial reprodutivo para a aplicação de técnicas dereprodução assistida. Foram submetidos a eletroejaculação 14 exemplares machos, adultos de Leão Africano, utilizando-se o método da eletroejaculação. Logo após a colheita, cada uma das 13 amostras obtidas foi examinada quanto ao volume, pH e aspecto geral, seguida pela avaliação da motilidade, movimento progressivo, índice demotilidade espermática e concentração. Uma alíquota do ejaculado foi fixada em solução de formol-salina a 10% e analisado em câmara úmida ao microscópio de interferência de fase, para o estudo dos aspectos morfológicos, classificando as alterações dos espermatozóides em defeitos maiores e menores. Os resultados foram: volume 5,83 ±3,35 ml; concentração 11,62 ± 14,51 x 106 espermatozóides/ml; motilidade total 73,85 ± 11,02 %; motilidade progressiva 3,35 ±0,63; índice de motilidade espermática 70,42 %, pH 8,1 ± 0,5; defeitos maiores 38,12 ± 19,41 %; defeitos menores 17,43 ± 10,36 %; defeitos totais 55,55 ± 19,30 %. Os defeitos maiores mais freqüentes foram gota citoplasmática proximal, peça intermediária dobrada com gota citoplasmática e peça intermediária dobrada. A cauda dobrada foi o defeito menor mais encontrado.

The aim of this study was to analyse the spermogram of a group of captive African lions (Panthera leo), housed at the Fundação Parque Zoológico de São Paulo, as a subject to avaluate the reproductive potential of the animals, an important aspect to apply artificial reproductive techniques. After chemical restraining, semen from 14 adult male lions was obtained by electroejaculation and the samples were examined right after the collection. The characteristics analyzed were: volume, pH, general aspect, motility, progressive motility, spermatic motility index and concentration. One aliquot of the samples were fixed in saline-formol solution 10% and examined under Phase Contrast Microscopy for morphological evaluation, classifying in minor and major defects. The results were: volume 5,83+/-3,35 ml; concentration11, 62 +/- 14,51 x 106 sperm cells/ml; total motility 73,85 +/- 11,02 % ; progressive motility 3,35 +/- 0,63; spermatic motility index 70,42 %, pH 8,1 +/- 0,5; major defects 38,12+/- 19,41 %; minor defects 17,43 +/- 10,36 %; total defects 55,55 +/- 19,30 %. The more frequent major defects were: proximal droplet, bent midpiece with cytoplasmic droplet and bent midpiece. Bent tailwas the most frequent minor defect observed.

Avaliação da morfologia e da compactação cromatínica em espermatozóides de galo (Gallus gallus, Linnaeus, 1758) através de microscopia eletrônica de transmissão / Morphology and chromatin condensation evaluation in fowl spermatozoa (Gallus gallus, Linnaeus, 1758) through the transmission electron microscopy

O espermograma é um dos principais métodos de avaliação da fertilidade em mamíferos, contudo, algumas alterações do sêmen como a compactação da cromatina dos espermatozóides, não são identificadas nessa rotina, podendo interferir significativamente na fertilidade de reprodutores. Na avicultura, geralmente a avaliação de reprodutores é feita por amostragem e os parâmetros avaliados são em menor número que em mamíferos, sendo que a compactação da cromatina dos espermatozóides de galo nunca foi explorada. Este trabalho teve como objetivo verificar através da microscopia eletrônica de transmissão, se existe diferentes intensidades de compactação de cromatina em espermatozóides de galo. Para isto, foram coletadas 20 amostras de sêmen de galo, que foram fixadas por 48 horas em glutaraldeído 4% tamponado em cacodilato de sódio 0,1M a pH 7,2, após centrifugado e lavado em tampão cacodilato o sêmen foi pós-fixados em tetróxido de ósmio a 1% mais ferrocianeto de potássio a 1,25%. O sedimento foi incluído, cortado e posteriormente contrastado. Os cortes foram examinados e documentados em microscópio eletrônico Zeiss EM-109. Geralmente, os espermatozóides de galo possuem acrossoma com material homogêneo ou levemente granular e de densidade moderada, o núcleo com cromatina densa e levemente granular, varia as tonalidades de cinza, ou seja, os graus de compactação. Observou-se também o “perforatorium”, estrutura que une o núcleo ao acrossoma. A inserção da cauda é por meio de dois centríolos, sendo um transversal e o outro longitudinal ao eixo. A peça intermediária possui axonema típico envolto por mitocôndrias aparentemente dispostas longitudinalmente. A transição entre as peças intermediária e principal da cauda é marcada pela ausência das mitocôndrias. A peça principal é formada pelo axonema envolto por uma camada de citoplasma granular. Na avaliação das patologias morfológicas observaram-se cabeças arredondadas, dobradas, envoltas por conteúdo da peça intermediária...

The spermiogram is one of the principal methods of evaluation of the fertility in mammals; however some semen modifications as the spermatozoa chromatin condensation, are not identified in this routine, and can expressively interfere in the fertility of reproducers. In poultry breeding, generally the evaluation of reproducers is made by sampling and the evaluated parameters are in fewer quantities than in mammals, considering the spermatozoa chromatin condensation of the fowl has never been exploited. The objective of this work is verifying through the transmission electron microscopy, whether there are different intensities of spermatozoa chromatin condensation of fowl. So that, have been collected 20 samples of fowl semen, which were placed in 4% glutaraldehyde buffered in 0.1M sodium cacodilate to pH 7.2 for 48 hours, after being centrifugated and rinsed in cacodilate tampon, the semen was post-fixed in 1 % osmium tetroxide plus 1.25% potassium ferrocyanide. The sediment was embedded, cut and contrasted. The thin sections were examined and documented in transmission electronic microscopy Zeiss EM -109. Generally the fowl spermatozoon has acrosome with homogenic or slightly granular and of moderate density material, the nucleus with dense or slightly granular chromatin, varies the gray scales, that is, the grades of condensation. It was also observed the "perforatorium", structure that links the nucleus to the acrosome. The insertion of the tail is made through two cetriules, being one transversal and the other longitudinal to the axle. The intermediate piece has typical axoneme involved by mitochondrias, apparently displayed longitudinally. The transition between the principal and the intermediate pieces of the tail is marked by the absence of the mitochondrias. The principal piece is formed by axoneme involved by a granular cytoplasm layer. In the evaluation of the morphological pathologies, it was observed rounded heads, folded, involved by the...

Effect of ribavirin on epididymal sperm count in rat.

Ribavirin (1-beta-D-ribofuranosyl-1, 2, 4-triazole-3-Carboxamide) is a potent inhibitor of inosine monophosphate dehydrogenase, used widely as an antiviral drug. Although it has been reported as a teratogen, its effect on spermatogenesis is not known. Male Wistar rats were segregated into 24 groups of 5 in each. Six groups were treated with water, 6 groups with 20 mg/kg, another 6 groups with 100 mg/kg and remaining 6 groups with 200 mg/kg for 5 days at intervals of 24 h (i.p.). Animals were anaesthetized at 14, 28, 35, 42, 70 and 105 days following the last exposure, laparatomy was conducted, epididymis was removed, minced in 1 ml phosphate buffered saline (PBS, pH 7.2), filtered and stained with 1% aqueous eosin Y. An aliquot was taken in haemocytometer, diluted in PBS and charged into Neubauer's chamber. Spermatozoa were counted in 8 squares except the central, and multiplied by 5 x 10(4). Data were analysed by Mann-Whitney "U" test. Ribavirin significantly decreased the sperm count in a dose and time dependent pattern and showed a recovery by day 105 except at 200 mg/kg. Ribavirin is reversibly cytotoxic to germ cells and decreases the production of spermatozoa.

Intra-uterine insemination versus fallopian tube perfusion with husband washed sperm as a method of management of oligospermia-induced infertility: a preliminary study

This study included 50 couples during the period from March 1993 till January 1996. All were suffering from primary infertility one to six years with no apparent cause, except male factor in the form of oligospermia with sperm count <20 millions/ml. The age of the male partner ranged between 28 and 42 years, while of the female partner ranged between 26 and 38 years. They were divided into two groups. The first group [n=28] have done 40-treatment cycle. While the second group [n=22] have done 28-treatment cycle. There was no significant difference between the two groups. All female partners were given the same induction protocol using CC and hMG and follow up by folliculometry and E2 plasma level. Once, one or more follicle reached size >/17 mm, 5000 IU hGG was given i.m. Thirty-three hours after administration of hCG, patients were randomly assigned to either IUI with 0.5 ml washed sperm [group I] or FSP with 4 ml husband washed sperm [group 2]. It was concluded that, FSP is a promising technique, which needs further study on a large group of patients suffering from various causes of infertility

Sperm motility and morphology as changing parameters linked to sperm count variations.

Variations in semen analyses of 177 males over a 1 year period were assessed. The average means of total counts, motility, morphology, total motile count and non-motile % were determined for 5 classes of patients ranging from azoospermic to normospermic. Positive relationships between a falling sperm count, a decrease in motility and total motile counts were seen. Also, increasingly, abnormal forms were found with lower sperm counts.

Evaluation of the Hamilton-Thorn Motility Analyzer for routine semen analysis.

In routine semen analysis of 102 patients the values of sperm concentration and the percentage of sperm motility were measured with the Hamilton-Thorn Motility Analyzer (HTMA) and compared to the data obtained by conventional semen analysis according to the guidelines of the World Health Organization (WHO). Results were classified according to the conventional semen analysis before statistical analyses. Overall, sperm concentration by both methods showed a correlation of 0.95 (p < 0.0001) but the HTMA gave higher values for sperm concentration (mean difference 10.8 +/- 18.9 x 10(6)/ml, mean +/- S.D.). Only a sperm concentration of between 40 and 80 x 10(6)/ml can be measured accurately with the HTMA. The percentage of sperm motility by both methods showed a correlation of 0.759 (p < 0.0001) but were estimated slightly lower by the HTMA than by conventional analysis (mean difference 1.7 +/- 15.8%). In conclusion, the HTMA system can not replace conventional semen analysis in routine semen analysis.

Evaluation of semen quality in a local Libyan population.

Semen analyses was carried out in a population of 1250 randomly selected Libyan males. Two semen samples collected from each volunteer were subjected to the routine analyses following the World Health Organization recommendations. The Libyan population had a higher percentage of men with sperm density in the range of 40-60 millions/ml. The percentage of men with sperm counts above 100 millions/ml is comparatively lower than that is reported by MacLeod and Gold (9). The seminal plasma transferrin levels showed a positive correlation with sperm density and alpha-glucosidase activity with sperm motility.