Diluente Tris suplementado com óleo de Mauritia flexuoxa sobre a qualidade do sêmen caprino após a criopreservação / Tris diluent supplemented with Mauritia flexuoxa oil on the quality of goat semen after cryopreservation

Objetivou-se verificar os efeitos, nos parâmetros espermáticos, na integridade mitocondrial, acrossomal e de membrana em células espermáticas, desencadeados pelo uso do Tris (Tris hidroximetil aminometano) suplementado com óleo de Mauritia flexuoxacomo diluente para criopreservação de sêmen caprino. Quatro caprinos clinicamente saudáveis foram utilizados. Os animais eram alimentados diariamente com volumoso (Pennisetum purpureum, Schum.), concentrado (ração peletizada com teor de 20% proteína, 300 g/animal/dia) e sal mineral específico para Caprinos (Caprinofós®), à vontade. Dois ensaios foram realizados: I ­ Teste de toxicidade; II ­ Criopreservação do sêmen com concentrações ideais. No teste de toxicidade as concentrações avaliadas foram: 5%, 10%, 15% e 20% de diluente a base de óleo de Mauritia flexuoxa. Após o teste de toxicidade, foi escolhido a concentraçãoque apresentou o melhor resultado (5%). Logo após, foram realizadas mais 32 coletas, que foram diluídas em Tris-gema-glicerol (grupo controle) ou diluente contendo óleo vegetal (Mauritia flexuoxa). As amostras foram criopreservadas com auxílio do aparelho Tk3000®. Após o período mínimo de uma semana as palhetas foram descongeladas em banho-maria a 37 °C por 30 segundos, acondicionadas em microtubos de centrifugação e homogeneizadas para a análise imediata de motilidade, vigor espermático e morfologia. Em seguida, por meio de sondas fluorescentes foram avaliadas a integridade de acrossomo, membrana plasmática (Diacetato de Carboxifluresceína e Iodeto de Propídeo) e função mitocondrial sob microscopia de epifluorescência. Quanto a motilidade e vigor, integridade mitocondrial e acrossomal, o grupo buriti foi inferior ao grupo controle. O Tris suplementado com óleo de Mauritia flexuoxa na concentração de 5% não influenciou significativamente a qualidade espermática, porém, observou-se morfologia e integridade de membrana favoráveis. Dessa forma, sendo uma alternativa para substituição de diluentes a base de produtos de origem animal.

The objective was to verify the effects, sperm parameters, mitochondrial, acrosomal and membrane integrity in sperm cells, triggered by the use of Tris (Tris hydroxymethyl aminomethane) supplemented with Mauritia flexuoxa oil as a diluent for cryopreservation of goat semen. Four goats clinically healthy were used. The animals were fed daily with bulky (Pennisetum purpureum, Schum.), concentrate (pelleted feed with 20% protein content, 300 g / animal / day) and mineral salt Specific for Goats (Caprinofós®), ad libitum. Two tests were carried out: I - Toxicity test; II - Semen cryopreservation with ideal concentrations. In the toxicity test as selected were: 5%, 10%, 15% and 20% of Mauritia flexuoxa oil-based diluent. After the toxicity test, the concentration that showed the best result (5%) was chosen. Soon after, a further 32 samples were obtained, which were diluted in Tris-glycerol (control group) or diluent containing vegetable oil (Mauritia flexuoxa). The samples were cryopreserved using the Tk3000® machine. After a minimum of one week, the samples were thawed in a 37 ° C water bath for 30 seconds, packed in centrifugation microtubes and homogenized for immediate analysis of motility, sperm vigor and morphology. Then, by means of fluorescent probes, the integrity of the acrosome, plasma membrane (Carboxyflurescein diacetate and Propidium Iodide) and mitochondrial function under epifluorescence microscopy were evaluated. As for motility and vigor, mitochondrial and acrosomal integrity, the buriti group was inferior to the control group. Tris supplemented with Mauritia flexuoxa oil at a concentration of 5% did not significantly influence sperm quality, however, favorable motility, morphology and membrane integrity were observed. Thus, being an alternative to replace diluents based on products of animal origin.

Presencia de Ochrobactrum anthropi en muestras de semen / Presence of Ochrobactrum Anthropi in Semen Samples

Introducción: El semen es una mezcla compleja de fluidos y células que posee las condiciones adecuadas para albergar microorganismos, especialmente bacterias. Objetivo: Evaluar la presencia de bacterias en el semen de individuos normozoospérmicos asintomáticos para infecciones urogenitales. Métodos: Se realizó una secuenciación estándar posterior a la amplificación por PCR con el uso de los cebadores universales 27F y 1492R para identificación de bacterias, en 10 muestras de semen de voluntarios normozoospérmicos asintomáticos para infecciones urogenitales. Resultados: Se identificó a Ochrobactrum anthropi en 8 de las 10 muestras seminales evaluadas y a Haemophilus paraurethrae o Escherichia coli en los dos restantes. O. anthropi es una bacteria comensal, ampliamente distribuida en la naturaleza, especialmente en las fuentes de agua que, a pesar de su baja virulencia, ocasionalmente causa infecciones en individuos inmunocomprometidos. Conclusión: La alta frecuencia de O. anthropi en las muestras de semen de individuos normozoospérmicos asintomáticos para infecciones urogenitales puede asociarse a procesos de contaminación durante la recolección de la muestra, debido a la amplia distribución de esta bacteria, especialmente en las fuentes de agua(AU)

Introduction: Semen is a complex combination of fluids and cells that can harbor microorganisms, especially bacteria. Objective: To assess the presence of bacteria in semen samples from asymptomatic normozoospermic individuals, for urogenital infections. Methods: Standard sequencing after PCR amplification was performed with the use of the universal primers 27F and 1492R for bacterial identification, in 10 semen samples of asymptomatic normozoospermic volunteers for urogenital infections. Results: Thisidentified Ochrobactrum anthropi in 8 out of 10 samples assessed. In the remaining two samples, we identified Haemophilus paraurethrae and Escherichia coli. O. anthropi is a commensal bacterium, widely spread in nature, especially in water sources that, despite its low virulence, occasionally cause infections in immune compromised individuals. Conclusion: The high frequency of O. anthropi in semen samples from asymptomatic normozoospermic individuals, for urogenital infections can be associated with contamination during the collection of the sample, due to the wide distribution of this bacterium, especially in water sources(AU)

Molecular characterization of ureaplasmas isolated from reproductive tract of goats and sheep from Brazil

Abstract Ovine/caprine ureaplasmas have not yet been assigned a species designation, but they have been classified into nine serotypes. Herein ureaplasmas were searched for in 120 samples of vulvo vaginal mucous from sheep and 98 samples from goats at 17 farms. In addition, semen samples were collected from 11 sheep and 23 goats. The recovered ureaplasma were from sheep and goats from animals without any reproductive disorder symptoms, but not all animals presented positive cultures. In sheep, 17 (68%) cultures of vulvovaginal mucous were positive for ureaplasma and 11 (27%) samples of semen presented positive cultures in animals with clinical signs of orchitis, balanoposthitis or low sperm motility. In goats four ureaplasma isolates were obtained from vulvovaginal mucus, but the semen samples were all negative. The isolates were submitted to Pulsed-field gel electrophoresis methodology and their 16S rRNA genes were sequenced. Fifty percent of ureaplasma recovered from sheep allowed for PFGE typing. Eleven isolates showed eight profiles genetically close to the bovine ureaplasmas. The 16S rRNA gene sequencing showed differences or similarities of isolates from sheep and goats, and the reference strains of bovine and human ureaplasma. Four clinical isolates from sheep were grouped separately. The studied ureaplasma isolates showed to be a diverse group of mollicutes.

Identification of Vaginal Fluid Using Microbial Signatures / 法医学杂志

OBJECTIVES@#To investigate the specific microbial signatures in vaginal fluid.@*METHODS@#Vaginal fluid （16 samples）, saliva （16 samples）, feces （16 samples）, semen （8 samples）, peripheral blood （8 samples）, urine （5 samples）, and nasal secretion （4 samples） were collected respectively. The 16S rRNA genes of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were amplified. PCR production was detected via a 3130xl Genetic Analyzer.@*RESULTS@#The detected number of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were 15, 5, 8, 14, and 3 in all vaginal fluid samples, respectively. Lactobacillus crispatus and Lactobacillus jensenii existed specifically in vaginal fluid.@*CONCLUSIONS@#There is a potential application value to detect Lactobacillus crispatus and Lactobacillus jensenii for the identification of vaginal fluid.

Real-time PCR for detection of Brucella ovis and Histophilus somni in ovine urine and semen / PCR em tempo real para detecção de Brucella ovis e Histophilus somni em urina e sêmen ovino

A epididimite infecciosa ovina é uma das principais enfermidades reprodutivas de carneiros. O presente estudo teve por objetivo desenvolver protocolos de PCR em tempo real para B. ovis e H. somni e avaliar sua aplicabilidade em amostras de sêmen e urina de carneiros. Delinearam-se primers e sondas espécie-específicos para cada agente. As sondas foram delineadas com o sistema TaqMan incorporando um marcador FAM para B. ovis e Cy5 para H. somni na extremidade 5' e um quencher na extremidade 3'. A PCR em tempo real para B. ovis e H. somni foi altamente sensível, uma vez que a amplificação de DNA ocorreu com até 0,2ng de DNA/reação. A especificidade dos iniciadores e sondas foi avaliada com amostras de DNA de outros agentes causadores de epididimite ovina e nenhuma amplificação inespecífica foi observada. A aplicabilidade da técnica em amostras biológicas também foi confirmada, pois não houve perda de eficácia (P>0,05) quando comparada à PCR convencional com amostras de sêmen e urina de carneiros experimentalmente infectados.

Efecto de los factores solubles de Staphylococcus aureus, Staphylococcus capitis y Staphylococcus epidermidis sobre la funcionalidad espermática / Effect of soluble factors of Staphylococcus aureus, Staphylococcus capitis and Staphylococcus epidermidis on sperm functionality

ANTECEDENTES: La interacción entre los espermatozoides con algunas especies bacterianas o sus factores solubles influyen en el deterioro de la calidad seminal, alterando la función reproductiva del hombre. OBJETIVO: El objetivo de este trabajo fue determinar el efecto de los factores solubles de Staphylococcus aureus, Staphylococcus capitis y Staphylococcus epidermidis sobre la calidad seminal. MÉTODO: Los factores solubles producto del metabolismo bacteriano de las cepas de S. aureus y S. Capitis sensible a oxacilina y S. aureus y S. Epidermidis resistente a oxacilina se incubaron con las muestras de semen de 20 voluntarios y se cuantificaron los parámetros seminales convencionales y funcionales por microscopía y citometría de flujo, respectivamente. RESULTADOS: Se observó una disminución en la movilidad espermática con los factores solubles de S. aureus, esta disminución fue mayor con la cepa sensible y el efecto negativo sobre la movilidad fue inmediato. Al incubar los espermatozoides con los factores solubles de S. aureus sensible a oxacilina, se afectaron todos los parámetros funcionales excepto la integridad de la cromatina y se observó menor liberación de especies reactivas de oxígeno; con los factores solubles de la cepa de S. aureus resistente a oxacilina se observó una disminución en la lipoperoxidación de membrana y en la expresión de anexina V. CONCLUSIÓN: Este estudio da cuenta del efecto negativo de los factores solubles de la bacteria S. aureus tanto sensible como resistente a oxacilina sobre los parámetros espermáticos convencionales y funcionales, y por ende en su función reproductiva.

BACKGROUND: The interaction between sperm with some bacteria species and their soluble factors are the deterioration of semen quality by altering the reproductive function of man. AIM: The aim of this study was to determine the effect of soluble factors Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus capitis on semen quality. METHODS: The soluble factors product of bacterial metabolism of the strains of S. aureus and S. capitis methicillin sensitive and S. aureus and S. epidermidis resistant to oxacillin, were incubated with semen samples from 20 volunteers. Subsequently, conventional seminal parameters were measured and functional quantified by microscopy and flow cytometry, respectively. RESULTS: A decrease was observed in sperm motility with soluble factors of S. aureus, this decrease was higher with the sensitive strain that with oxacillin resistant strain and the negative effect on motility was immediate. By incubating the sperm with soluble factor from oxacillin-sensitive S. aureus, all functional parameters were affected except the chromatin integrity and reduced release of reactive oxygen species, mean fluorescence intensity in oxacillin resistant S. aureus strain was decrease in membrane lipid peroxidation and annexin V expression. CONCLUSIONS: This study reports the negative effect of soluble factors of bacteria either S. aureus sensitive and resistant to oxacillin, over conventional and functional sperm parameters, and therefore in their reproductive function.

Espermocultivo: crecimiento bacteriano del eyaculado y su relación con los parámetros seminales / Spermoculture: bacterial growth of the ejaculate and its relationship with the seminal parameters

ANTECEDENTES: En el semen, algunos microorganismos pueden encontrar las condiciones óptimas para sobrevivir, ocasionando daños a los espermatozoides y desencadenando procesos de infertilidad o infecciones del tracto reproductivo. Entender el papel de los microorganismos aislados en el semen, contribuye a mejorar el diagnóstico de casos de infertilidad donde la única causa aparente son los procesos infecciosos. OBJETIVO: Describir y correlacionar los parámetros seminales y el crecimiento bacteriano del eyaculado. MÉTODOS: Identificación de los microorganismos aislados en 43 espermocultivos-clínicos y 28 espermocultivos-investigación. Se realizó conteo de las unidades formadoras de colonia a los espermocultivos-investigación y análisis de las características espermáticas. Resultados: Se obtuvo crecimiento bacteriano en 14 (32,6%) de los espermocultivos-clínicos y 15 (53,6%) de los espermocultivos-investigación. Los microorganismos aislados fueron Enterococcus faecalis, Escherichia coli, Morganella morganii, Staphylococcus spp coagulasa negativo, Klebsiella pneumoniae y microbiota mixta. En este estudio se observó abundante crecimiento de cocos aerobios. Finalmente, no se encontró asociación entre la disminución en la calidad de los parámetros seminales y los microorganismos. CONCLUSIONES: La presencia de bacterias en el semen no afecta la calidad seminal.

BACKKGROUND: Microorganisms can find the optimal conditions for survival in semen, causing damage to the spermatozoa and triggering processes of infertility or reproductive tract infections. Therefore, understanding the role of the microorganisms present in semen can help to improve the diagnosis of infertility cases where the only apparent cause is infectious processes. OBJECTIVE: To describe and correlate semen parameters and bacterial growth in ejaculate. METHODS: Identification of microorganisms isolated in 43 clinical spermocultures and 28 research spermocultures. We assessed colony-forming unit counts and sperm characteristics of research spermocultures. In addition, semen parameters were evaluated in each ejaculate. RESULTS: Bacterial growth was obtained in 14 (32.6%) of the 43 clinical spermocultures and 15 (53.6%) of the 28 research spermocultures. The isolated microorganisms were Enterococcus faecalis, Escherichia coli, Morganella morganii, Staphylococcus coagulase negative, Klebsiella pneumoniae and mixed microbiota. Finally, in this study a large growth of aerobic cocci was observed. We did not find association between the decline in the quality of semen parameters and microorganisms. CONCLUSION: The presence of bacteria in semen does not affect semen quality.

Detection of Ureaplasma spp. in semen samples from sheep in Brazil

A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples.

Micoplasma hominis, Ureaplasma urealyticum y bacterias aeróbicas en el semen de hombres que consultan por infertilidad / Micoplasma hominis, Ureaplasma urealyticum and aerobic bacteria present in the semen from men attending infertility service

Introducción: las infecciones en el semen humano pueden alterar la calidad espermática, y vincularse con problemas de infertilidad masculina. Objetivo: determinar la frecuencia de infecciones por Micoplasma hominis, Ureaplasma urealyticum y bacterias aeróbicas en el semen de hombres que consultan por infertilidad, e identificar si existe relación entre las infecciones encontradas y las alteraciones en las variables de calidad del semen. Métodos: se realizó un estudio descriptivo transversal, para evaluar muestras de semen de 140 hombres, con edades entre 20 y 45 años, provenientes de las consultas de infertilidad del Instituto Nacional de Endocrinología. Se realizó un espermograma completo, que incluyó leucocitospermia, siguiendo los lineamientos de la OMS, para determinar las variables cualitativas y cuantitativas del semen. Las muestras de semen fueron cultivadas en agar sangre y agar chocolate a 37° C en atmósfera de CO2 para investigar bacterias aeróbicas, y se utilizó un juego de reactivos (Mycoplasma System Plus) que permite realizar el cultivo, la identificación, el conteo semicuantitativo y el antibiograma de micoplasmas/ureaplasma urogenitales. Se tuvo en cuenta los aspectos éticos, y los resultados obtenidos se analizaron mediante cálculo de por cientos y la aplicación de la prueba de chi cuadrado. Resultados: de las 140 muestras de semen evaluadas, 58 (41,4 por ciento) mostraron la presencia de infecciones, de ellas 37 correspondieron a Ureaplasma urealyticum (25,7 por ciento), 2 a Micoplasma hominis (1,4 por ciento) y 19 a bacterias aeróbicas (13,8 por ciento ). Al comparar las variables cualitativas y cuantitativas del semen con los sujetos infectados y no infectados, no se observaron diferencias estadísticamente significativas en ninguna de las variables de calidad espermática evaluadas. Conclusiones: la frecuencia total de infecciones, en la muestra estudiada, fue relativamente alta, pero no asociada a alteraciones en las variables seminales(AU)

Introduction: human semen infections can alter the sperm quality and be associated to male infertility disorders. Objectives: to determine the frequency of infections from Micoplasma hominis, Ureaplasma urealyticum and other aerobic bacteria in the semen of men who attended the infertility service, and to identify whether there is some relation between the detected infections and the altered semen quality variables or not. Methods: a cross-sectional descriptive study was performed to evaluate semen samples from 140 men aged 20 to 45 years, who attended the infertility service at the National Institute of Endocrinology. According to the WHO guidelines, a complete spermiogram including leukocytospermia was performed in order to determine the qualitative and quantitative variables in the semen. The semen samples were cultured in blood agar and in chocolate agar at 37oC under CO2 environment to find out possible aerobic bacteria. To this end, a set of reagents known as Mycoplasma System Plus was used, allowing the culture, the identification, the semi-quantitative count and the antibiogram of urogenital mycoplasms/ureaplasms. The ethical aspects were allowed for; the results were analyzed through percentage estimations and the chi square test. Results: out of the 140 evaluated semen samples, 58 (41.4 percent) showed some infection, 37 of them were caused by Ureaplasma urealyticum (25.7 percent), 2 by Micoplasma hominis (1.4 percent) and 19 by the aerobic bacteria (13.8 percent). When making a comparison of the qualitative and quantitative variables of the semen from infected and non-infected subjects, there were not any statistically significant differences in the evaluated variables of the sperm quality. Conclusions: the total frequency of infections in the studied sample was relatively high, but was not associated to altered seminal variables(AU)

Infecciones de transmisión sexual en semen: el hombre como vector de transmisión / Sexual transmission infections in semen: men as vector transmission

En los últimos años el estudio de las infecciones de transmisión sexual ha cobrado gran importancia debido principalmente al incremento de estas en parejas heterosexuales y hombres que tienen sexo con hombres. En mujeres existe mucha información de epidemiología y patogénesis de estas infecciones, sin embargo, en hombres la información es muy escasa debido a que la mayoría no presenta sintomatología. En los últimos años se ha evidenciado un creciente interés en el estudio del semen como vía de transmisión, debido principalmente a la afinidad de algunos patógenos con los espermatozoides. Dentro de los principales microorganismos infectantes en semen se encuentran Chlamydia trachomatis, Neisseria gonorrhoeae, Mollicutes, Virus de la Inmunodeficiencia Humana tipos 1 y 2, Virus Herpes Simplex 1 y 2, Virus Papiloma Humano, Virus de la Hepatitis B y C, Citomegalovirus, Virus Epstein-Barr y Trichomonas vaginalis.

Sexually transmitted infections study has become an important issue in these days, mainly due to the increment of heterosexual and men have sex with men partners of people. In women, there is a lot information about epidemiology and pathogenesis of these infections. However, the information is very limited in men, because most infected men are asymptomatic. In recent years, there has been an increasing interest in study of semen as a transmission way, due to the affinity of some pathogens to sperm. The most prevalent microorganisms infecting semen are: Chlamydia trachomatis, Neisseria gonorrhoeae, Mollicutes, Human Immunodeficiency Virus Types 1 and 2 Herpes Simplex Virus 1 and 2, Human Papillomavirus, Hepatitis B and C virus, Cytomegalovirus, Epstein-Barr and Trichomonas vaginalis.

Seminal bacterial contaminations: probable factor in unexplained recurrent pregnancy loss

It is estimated that about 50% of causes of recurrent pregnancy loss [RPL] cases remain unknown. Sperm factors are suggested to have probable role in cases with RPL. The goal was to determine the possible relationship between semen bacterial contaminations with unexplained RPL. Also, the correlation between number of bacterial colony and sperm chromatin condensation was examined This study consisted of 30 fertile men [group A] and 30 infertile [group B] men with unknown RPL. Semen collection and analysis were done according to WHO manuals. Sperm count and motility were evaluated by Makler chamber. Eosin-Nigrosin and Papanicolaou staining methods were applied for viability and morphology assessment, respectively. The semen samples from both groups were cultured for aerobic bacteria. Aniline blue [AB] and toluidine blue [TB] staining methods were applied for evaluating sperm chromatin condensation. The numbers of colonies were significantly higher in group B when compared to group A. Also, S. aureus and E. coli contaminations showed significant differences between two groups. Both AB+ and TB+ sperm cells showed significant increase in group B compared to group A. There was a significant negative correlation between colony number and progressive motility [p=0.01], and sperm viability [p=0.007]. In addition, positive correlations were found between colony number and AB+ [p=0.001] and TB+ [p=0.004] as well. Bacterial contaminations in semen of men from RPL couples had significantly higher levels when compared to fertile controls. Presence of microorganisms in semen may be correlated with irregular sperm parameters and quality

Mycoplasmas y anticuerpos anti-Chlamydia en semen de hombres infértiles y su relación con la calidad seminal y los marcadores de glándulas sexuales accesorias masculinas / Mycoplasmas and antibodies anti-Chlamydia in semen of infertile men and their relationship with seminal quality and markers of male accessory sex glands

La infertilidad masculina puede deberse a inflamación o infección del tracto genital entre otras causas. En el problema de la infertilidad masculina pueden estar implicadas las glándulas sexuales accesorias y la función espermática. En este trabajo se trata de asociar los gérmenes más frecuentes en semen de hombres infértiles incluyendo Chlamydia trachomatis, Mycoplasma hominis y Ureaplasma urealyticum con las características seminales, y los niveles de fructosa, ácido cítrico y a-glucosidasa neutra como marcadores de las glándulas sexuales accesorias masculinas. La detección de los anticuerpos indicó que C. trachomatis fue el germen de mayor prevalencia. Los anticuerpos (Acs) anti-Chlamydia, Mycoplasma hominis y Ureaplasma urealyticum se asociaron con descenso de los marcadores glandulares fructosa y a-glucosidasa. Por otra parte se observó aumento del pH y leucocitospermia en los pacientes con Acs anti-Chlamydia. La evaluación microbiológica y la bioquímica del semen orientarían más sobre la propagación de la infección y permitiría seleccionar la terapia más efectiva. Se observó que es importante la evaluación microbiológica y de los marcadores de glándulas accesorias sexuales masculinas en el semen para diagnosticar y tratar las infecciones masculinas.

Male infertility may be due to inflammation or infection of the genital tract among other causes. Male accessory sex glands and sperm function may also be involved in the problem of infertility. This study tries to associate the most frequent bacteria in semen of infertile men including Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum with the seminal characteristics and levels of fructose, citric acid and a-neutral glucosidase as markers of the accessory glands. Detection of antibodies anti Chlamydia trachomatis indicated that it was the most prevalent germ. Antibodies (Ab) anti-Chlamydia, Mycoplasma hominis and Ureaplasma urealyticum were associated with a decrease of the glandular markers fructose and a-neutral glucosidase. On the other hand, there were increased pH and leukocytospermia in men positive for antibodies anti-Chlamydia. Microbiological and biochemical evaluation of semen could orient more about the spread of infection and allow for the selection of the most effective therapy. We find that microbiological and glandular accessory markers assessments in semen are important to diagnose and to treat infections.

Isolamento de Corynebacterium pseudotuberculosis no sêmen de um carneiro na região central do Rio Grande do Sul / Isolation of Corynebacterium pseudotuberculosis from the semen of a sheep in the central region of Rio Grande do Sul, Brazil

O presente trabalho descreve o isolamento do Corynebacterium pseudotuberculosis no sêmen de um carneiro da raça Suffolk proveniente do Município de São Martinho da Serra, RS. Ao exame físico constatou-se aumento de volume subcutâneo aderido à pele, junto à inserção do escroto, próximo ao plexo pampiniforme esquerdo. Realizou-se exame andrológico com coleta de sêmen e, posteriormente, punção no local da lesão, obtendo-se material viscoso de coloração amarelo esverdeado. As amostras coletadas (sêmen e material obtido da punção) foram enviadas ao LABAC (Laboratório de Bacteriologia Veterinária da Universidade Federal de Santa Maria) para realização de cultura bacteriana. Em ambas as amostras, foi isolado o cocobacilo Gram positivo denominado C. pseudotuberculosis.

The present paper describes the isolation of Corynebacterium pseudotuberculosis from a semen sample of a Suffolk ram from São Martinho da Serra County, state of Rio Grande do Sul, Brazil. Physical examination revealed a subcutaneous mass close to the insertion of the scrotum, more specifically around the left pampiniform plexus. A routine andrological exam with semen collection was performed. Also, the mass was aspirated yielding a viscous greenish material. Samples were sent to a bacteriology lab for culture and identification. Gram positive coccobacillus C. pseudotuberculosis was isolated from both samples.

Impact of infection on the secretory capacity of the male accessory glands

INTRODUCTION: Studies that compare the impact of different infectious entities of the male reproductive tract (MRT) on the male accessory gland function are controversial. MATERIAL AND METHODS: Semen analyses of 71 patients with proven infections of the MRT were compared with the results of 40 healthy non-infected volunteers. Patients were divided into 3 groups according to their diagnosis: chronic prostatitis NIH type II (n = 38), chronic epididymitis (n = 12), and chronic urethritis (n = 21). RESULTS: The bacteriological analysis revealed 9 different types of microorganisms, considered to be the etiological agents, isolated in different secretions, including: urine, expressed prostatic secretions, semen and urethral smears: E. Coli (n = 20), Klebsiella (n = 2), Proteus spp. (n = 1), Enterococcus (n = 20), Staphylococcus spp. (n = 1), M. tuberculosis (n = 2), N. gonorrhea (n = 8), Chlamydia tr. (n = 16) and, Ureaplasma urealyticum (n = 1). The infection group had significantly (p < 0.05) lower: semen volume, alpha-glucosidase, fructose, and zinc in seminal plasma and, higher pH than the control group. None of these parameters was sufficiently accurate in the ROC analysis to discriminate between infected and non-infected men. CONCLUSION: Proven bacterial infections of the MRT impact negatively on all the accessory gland function parameters evaluated in semen, suggesting impairment of the secretory capacity of the epididymis, seminal vesicles and prostate. These findings were associated with an infectious related significant increase of semen pH. None of the semen parameters evaluated can be suggested as a diagnostic tool for infection.

Semen and urine culture in the diagnosis of chronic bacterial prostatitis

OBJECTIVE: To assess the diagnostic accuracy of semen and urine culture in the diagnosis of chronic bacterial prostatitis (CBP). MATERIALS AND METHODS: In 70 consecutive men suspected of having chronic bacterial prostatitis along with 17 asymptomatic controls, we obtained urine and semen cultures followed 1 week later by the Meares and Stamey test, our reference standard. The interpretation of each of the cultures was blind to the results of other tests. RESULTS: 139 men were referred for evaluation of chronic bacterial prostatitis and 70 received all tests. Additionally, 17 control men volunteered to participate. The Meares and Stamey Test was positive in 69 (79 percent) patients. The semen culture had a sensitivity of 45 percent and a specificity of 94 percent. The likelihood ratio associated with a positive semen culture was 8.1 (95 percent confidence interval (CI) 1.2 to 55.3); the likelihood ratio associated with a negative semen culture was 0.6 (95 percent CI 0.5 to 0.7). The urine culture had a sensitivity of 4 percent and a specificity of 100 percent. The likelihood ratio of a positive urine culture was infinity and of a negative urine culture was 0.96 (95 percent CI 0.9 to 1). CONCLUSIONS: While a positive semen culture in a symptomatic patient may suffice to select and start antibiotic treatment against chronic bacterial prostatitis, a negative culture does not rule out the condition. Urine cultures alone are not useful for diagnosing CBP. The Meares and Stamey test remains important for the diagnosis of CBP in practice.

O exame do sêmen na infertilidade masculina: V-exame microbiológico / The microbiological examination of the sêmen to evaluate male infertility

Neste artigo, o autor revisa o papel do exame microbiológico do sêmen para avaliar infertilidade masculina. São descritos os efeitos negativos de bactérias aeróbicas e anaeróbicas, micoplasmas, fungos, Trichomonas sp., Chlamydia trachomatis, Gardnerellavaginalis e Mobiluncus sp. na qualidade do sêmen. A influência de leucocitospermia e bacteriospermia também é revisada. Simultaneamente,também são feitas algumas considerações sobre metodologias atualmente usadas para investigar estes microrganismos no sêmen.

In this article, the author reviews the role of the microbiological examination of the semen to evaluate male infertility.They are described negative effects of aerobic and anaerobic bacteria, mycoplasmas, fungi, Trichomonas sp., Chlamydia trachomatis, Gardnerella vaginalis and Mobiluncus sp. on the semen quality. Influence of leukocytosp.ermia, and bacteriosp.ermia are also reviewed. Simultaneously, they are also made some considerations about methodologies currently used to investigate these microorganismson semen.

Detection of M. tuberculosis in clinical samples of diversified nature by IS6110 based PCR.

Performance of the polymerase chain reaction technique based on IS6110 sequence was evaluated in clinical samples obtained from pulmonary and extrapulmonary cases of tuberculosis. One hundred and seventy two samples were processed for detection of M. tuberculosis by ZN stained smear examination, LJ medium culture, BACTEC radiometric culture and PCR tests amplifying 123bp region of IS6110 sequence. A significant difference was seen in the sensitivities of different tests, the figures being 83% for PCR test, 35.2% for smear examination, 47.16% for LJ culture and 53.45% for BACTEC culture (p < 0.05). However, no significant difference was found as far as specificity was concerned. PCR test sensitivity in. pulmonary and extrapulmonary clinical samples were 90.14% and 77.27% respectively and found to be significantly higher (p < 0.05) when compared with those of other tests. The mean detection time for M. tuberculosis was 24.03 days by LJ medium culture, 12.89 days by BACTEC culture and less than one day by PCR test. PCR based on IS6100 sequence is highly sensitive method for the early diagnosis of pulmonary and extrapulmonary tuberculosis.

Avaliação microbiológica do sêmen fresco e congelado de reprodutores caprinos / Microbiological evaluation of buck fresh and frozen semen

Objetivou-se avaliar a flora microbiana no sêmen fresco e congelado de reprodutores caprinos, assim como a eficácia dos antibióticos estreptomicina, penicilina e gentamicina, na viabilidade de doses de sêmen congeladas. Foram utilizados 25 reprodutores de diferentes raças, submetidos a duas colheitas de sêmen através do método da vagina artificial, após higiene da região prepucial. A primeira colheita do sêmen foi realizada visando o exame microbiológico e a segunda teve como objetivo proceder a congelação, após diluição em leite desnatado, utilizando penicilina + estreptomicina (A1), gentamicina(A2) ou sem antibiótico (A3). Ao proceder a avaliação microscópica no sêmen fresco, evidenciou-se média de 87,92 ± 7,76 % de motilidade individual progressiva (MIP) e 4,96 ± 0,20 de vigor espermático. Em relação à avaliação bacteriana, constatou-se principalmente bactérias do gênero Staphylococcus spp e Bacillus sp. Após a congelação do sêmen, não foram evidenciadas diferenças (P>0,05) entre os grupos quantoa MIP e vigor espermático. Entretanto, na avaliação microbiológicapós-descongelação, a bactéria do gênero Staphylococcus spp esteve presente na maioria das amostras. Observou-se também que a gentamicina (13,3mg/mL) apresentou melhor atividade antimicrobianano processo de congelação do sêmen, concluindo-se que pode ser o antibiótico usado na congelação do sêmen de reprodutores caprinos.

The aim of this research was to evaluate the microbial flora in the fresh and frozen semen of goat reproducers, as well as the effectiveness of the antibiotics estreptomicin, penicillin and gentamicin in cryopreservation of semen. It were used 25 males of different breeds, submitted to two semen collect through the artificial vagina method after cleanliness of prepucial region. The first collection of semen aimed the microbiological exam. The second collection had as goal accomplish freezing, after dilution in skimmed milk, with penicillin+ estreptomicin (A1), gentamicin (A2) or control (A3). After microscopic evaluation, it was evidenced average of 87.92 ± 7.76% of MIP and 4.96 ± 0.20 of spermatic vigor in fresh semen. Regarding bacterin evaluation, it was verified, mostly, bacteria of the gender Sthaphylococcus spp and Bacillus sp. After semen cryopreservation, it was observed that there wasn’t difference (P>0.05) among groups in MIP and spermatic vigor. However, in the microbiological evaluationof frozen-thawed semen, bacteria of Sthaphylococcus spp gender was present in great part of samples. Gentamicin (13.3mg/mL) promoted larger inhibition of the bacterial growth in the semen post-freezing, concluding that gentamicin can be the antibiotic used for freezing of goat semen.

Painful ejaculation; an underestimated sexual possible underlying serious etiology

Painful ejaculation, a previously an underestimated ejaculatory and sexual dysfunction, has gained increased attention being one of the significant sexual dysfunctions associating LUTS of BPH. Other underlying etiopathologic factors are not well studied, To study the underlying etiologic factors responsible for painful ejaculation in a group of patients having this symptom as the main complaint and the efficacy of different treatment modalities. Furthermore we want to test our hypothesis that in suspected cases of genital TB, it is better to search for it in semen rather than in urine. 60 male patients with painful ejaculation were enrolled in this study. Their mean age was 39,4 +/- 8.7 years while the mean duration of their complaint was 15.4 +/- 4.8 months. Patients were subjected to thorough history taking and physical examination including DRE. Patients were extensively investigated by urinalysis, urine culture, Ziehi Neelsen [Z,N.] staining and PCR for acid fast bacilli in both urine and semen ,semen culture, semen culture for TB, TRUS [ +/- biopsy] and/or cystoscopy when indicated in addition to PSA determination in all men above 50 years old. Associated symptoms were premature ejaculation, chronic prostatitis manifestations, ED, cystitis manifestations, infertility, partner dyspareunia or hemospermia in that order of frequency. Significant physical findings included BPH [15], epididymal mass[1], recurrent epididymoorchitis [1] scrotal sinus [1] and prostatic carcinoma [1], UTI was proved by urine culture in 20case with E-coli strains predominantly isolated. Urine PCR for TB was positive in 5 patients [13% sensitivity and 100% specificity] while same test in semen yielded astonishingly high incidence of TB in 40 patients [100% sensitivity and 91% specificity].Semen culture for TB confirmed its presence in 38 out of 40 PCR positive specimens. TRUS findings included calcular prostatitis [7], BPH [15], Prostatic adenocarcinoma [1], pathologic seminal vesicles [15],ejaculatory duct obstruction [3] and prostatic cysts [2]. Treatment of the underlying etiologies produced significant improvement of pain. Alpha blockers improved pain in 93% and 70% of BPH and chronic prostatitis patients respectively. Painful ejaculation may be an important indicator of a serious underlying disease such as prostatic carcinoma. In our locality genitourinary TB is prevalent among this group of patients. We introduce the application of PCR in semen as a highly sensitive and specific test which should be done whenever genitourinary TB is suspected as it showed better sensitivity than same test in urine. Alpha blockers proved effective in relieving painful ejaculation in BPH patients and to less extent in chronic prostatitis patients

Infección por Chlamydia trachomatis en varones y su asociación con las alteraciones ginecológicas de su compañera sexual / Chlamydia trachomatis infection in men and its association to gynecologic alterations of his sexual mate

Objective.To determinate the frequency of Chlamydia trachomatis infection in male partners of infertile couples who attend to the infertility clinic at Instituto Nacional de Perinatologia, as well as to compare the clinical data and lifestyle between C. trachomatis-inifected and uninfected men to establish a possible association with gynecological damage in their sexual female partners. Methods. An open prospective study was performed in infertile couples, whose follow up was carried out at Instituto Nacional de Perinatologia between June 2000 and April 2001. Urethral and cervical swabs were obtained from each couple and the specimens were subjected to a C. trachomatis-specific liquid-phase hibridization test (PACE-2) and routine microbiological analysis. Semen analysis were also included. A relative risk (RR) test was done to analyze variables and square chi test was used to analize clinical and gynecological data from female partners and data from semen examination. Statistical differences were considered as significant when the p value was below 0.05. Results. C. trachomatis active infection was found in 14 out of 384 urethral swabs (3.6%). No significant alterations were observed in semen samples of C. trachomatis-infected men, as compared to non-infected individuals. Microbiological analyses of semen showed a significant isolation o/Mycoplasma sp (RR = 5.87, IC95% 1.4-24.7). Eight out of fourteen female partners of C. trachomatis-infected men were also infected with C. trachomatis (RR= 10.57, IC95% 5.67-19.7), Candida albicans was other pathogen isolated from 8/14 of those women (RR = 1.89, IC95% 1.17-3.05). Gynecological and obstetrical associations found among female partners of C. trachomatis-infected men were as follows: tubal adhesions in 10/14 (RR = 1.54, IC95% 1.08-2.18), salpingitis in 2/14 (RR = 2.2), history of ectopic pregnancies in 11/14 (RR =2.94, IC95% 1.01-8.53) and abnormal pregnancy loss in 9/14 (RR = 1.5). Conclusion. A low prevalence of C. trachomatis infection was observed among male partners of infertile couples as compared with other reports, but this discrepancy could be attributable to the specimen collection and diagnostic assay used. Otherwise, this data suggests that a chronic pathogen's antigenic stimulation may result in an increased formation of tubal adhesions and/or in ectopic pregnancies among female partners of C. trachomatis-infected individuals. Thus, preventive and control measures must be introduced into men's healthcare services, through laboratory and clinical examination, since these subjects are the main reservoirs of C trachomatis.

Objetivo. Determinar la frecuencia de infección por Chlamydia trachomatis y comparar la información clínica y el estilo de vida de varones con y sin infección por este patógeno, así como su asociación con las alteraciones ginecológicas que presenta su compañera sexual en un grupo de parejas que asisten a la Clínica de Infertilidad del Instituto Nacional de Perinatologia de la Ciudad de México. Métodos. Se realizó un estudio abierto, longitudinal y prospectivo en un grupo de parejas con diagnóstico de infertilidad, que fueron tratadas en el Instituto Nacional de Perinatologia durante el periodo de junio del 2000 a abril del 2001. Se recolectaron muestras uretrales y cervicales de cada pareja para el diagnóstico de C. trachomatis mediante la prueba de hibridación en fase líquida (PACE-2). También se recolectaron muestras de semen para el análisis de espermatobioscopia y se hicieron cultivos microbiológicos de rutina a las muestras cervicales y de semen. Los datos microbiológicos, clínicos y ginecológicos de los participantes fueron comparados por %z, el análisis de tendencia para proporciones fue usado para establecer el nivel de riesgo en las variables (RR). Las diferencias fueron consideradas estadísticamente significativas si p < 0.05. Resultados. Se analizaron un total de 384 muestras uretrales de varones, 14 presentaron infección activa por C. trachomatis (3.6%), Los datos de espermatobioscopia de los individuos positivos a C. trachomatis no mostraron alteraciones significativas con respecto al de varones no infectados con esta bacteria. El análisis microbiológico del semen mostró un número de aislamientos significativos de infección por Mycoplasma sp. (RR = 5.87, IC95% 1.40-24.70). En cuanto a las muestras cervicovaginales de mujeres con compañero sexual infectado por C. trachomatis, los patógenos aislados con mayor frecuencia fueron: Candida albicans en ocho de 14 (RR = 1.89, IC95% 1.17-3.05) y C. trachomatis en ocho de 14 (RR = 10.57, IC95% 5.67-19.7). Las asociaciones ginecológicas y obstétricas de la compañera sexual de varones positivos a C. trachomatis fueron adherencias tubáricas en 10 de 14 (RR = 1.54, IC95% 1.08-2.18), salpingitis en dos de 14 (RR = 2.2), antecedentes de embarazos ectópicos en 11 de 14 casos (RR = 2.94, IC95% 1.01-8.53) y abortos previos en nueve de 14 (RR = 1.5). Conclusión. Se observó una baja prevalencia de infección por C. trachomatis en los varones de mujeres infértiles en comparación con lo reportado por otros autores, esta diferencia puede estar dada por el método de diagnóstico y la toma del producto. Estos resultados sugieren que el estímulo constante del patógeno produce un aumento de adherencias tubáricas y embarazos ectópicos en las compañeras sexuales de los varones infectados con C. trachomatis. Por lo que una evaluación diagnóstica y de laboratorio deberá ser llevada a cabo en el varón como una medida de prevención y control para la infección por este patógeno, ya que estos individuos actúan como reservónos importantes de infección.