Characterization of the Campylobacter fetus subsp. venerealis adhesion to bovine sperm cells / Caracterización de la adhesión de Campylobacter fetus subsp. venerealis a espermatozoides bovinos

Campylobacter fetus is extracellular bacteria of the genital tract of cattle. They cause infertility and abortion, but there is no documented information on the susceptibility of bovine sperm cells to this bacteria. The aim of this present work was to study the effects provoked by Campylobacter fetus subsp. venerealis when in interaction with bovine sperm cells. The bovine spermatozoa were obtained frozen bovine semen pooled from uninfected bulls, and were exposed to living campylobacter over different periods of time. Light microscopy and scanning electron microscopy first revealed a tropism, then a close proximity followed by tight adhesion between these two different cells. A decrease in the spermatozoa motility was observed. Motile bacteria were observed during the next 3 h, this process began with a tight membrane­membrane adhesion. The adhesion between Campylobacter fetus to the sperm cell occurred either by the flagella or by sperm head. Results from this study demonstrated with light microscopy scanning electron microscopy allowed us to characterize some aspects of the interaction of Campylobacter fetus subsp. venerealis and bovine sperm while preserving the cellular and bacterial structure. This ex vivo model might be useful for studies on adhesion and cytopathogenicity of different field strains of Campylobacter fetus.

Campylobacter fetus subsp. venerealis es un patógeno extracelular del tracto genital de bovinos. En las hembras causa subfertilidad y aborto, mientras que los toros son portadores en el esmegma prepucial y se desconoce si provoca daño en los espermatozoides. El objetivo del presente trabajo fue estudiar los efectos de Campylobacter fetus subsp. venerealis sobre espermatozoides bovinos. Los espermatozoides obtenidos a partir de pajuelas de semen pertenecientes a toros no infectados, se coincubaron con una cepa de Campylobacter fetus subsp. venerealis por diferentes períodos de tiempo. Por microscopía óptica y electrónica de barrido se observó el tropismo inicial de la bacteria hacia los espermatozoides y la adhesión bacteriana, de forma colateral se observó su efecto en el espermograma. Post incubación los espermatozoides presentaron menor motilidad progresiva y mayor porcentaje de muertos con respecto al control. Se comprobó la viabilidad de la bacteria a las 3 h. Se registró la adhesión de Campylobacter fetus subsp. venerealis a la membrana celular de distintas porciones del espermatozoide: cabeza, pieza media, cuello y cola. Los resultados de este estudio permitieron caracterizar la interacción entre Campylobacter fetus subsp. venerealis y espermatozoides bovinos por microscopía óptica y electrónica de barrido. La aplicación de este modelo ex vivo permitirá profundizar los conocimientos referentes a los procesos de adhesión y citopatogenicidad de Campylobacter fetus.

Evaluación in vitro del efecto de Neisseria gonorrhoeae y los factores solubles producto de su metabolismo sobre la calidad espermática / In vitro evaluation of the effect of Neisseria gonorrhoeae and the soluble factors produced by its metabolism on sperm quality

AANTEDENTES: N. gonorrhoeae es una de las principales causas de uretritis, prostatitis y epididimitis en hombres. En el tracto urogenital masculino, esta bacteria o los factores solubles producto de su metabolismo pueden interactuar con los espermatozoides y alterar su calidad. OBJETIVOS: Determinar el efecto de la incubación in vitro de N. gonorrhoeae y los factores solubles producto de su metabolismo sobre la calidad espermática y evaluar la interacción bacteria-espermatozoide. MÉTODO: Se realizó incubación de una concentración 0,5 McFarland de N. gonorrhoeae y de los factores solubles producto de su metabolismo bacteriano con muestras de semen de voluntarios aparentemente sanos. Se cuantificaron los parámetros espermáticos convencionales (movilidad y viabilidad) y funcionales (potencial de membrana mitocondrial, integridad y lipoperoxidación de la membrana espermática, detección de especies reactivas del oxígeno, integridad de la cromatina y expresión de Anexina V), empleando microscopía y citometría de flujo, respectivamente. Para evaluar la interacción de N. gonorrhoeae con los espermatozoides humanos se realizaron extendidos en placas para su observación al microscopio. RESULTADOS: N. gonorrhoeae puede unirse al espermatozoide y disminuir la viabilidad espermática luego de 1,5 horas de incubación con los espermatozoides humanos (84,5% vs 66,5%, p<0,05), sin afectar los parámetros espermáticos funcionales. CONCLUSIÓN: N. gonorrhoeae interactúa con los espermatozoides humanos afectando la viabilidad espermática.

BACKGROUND: N. gonorrhoeae is a major cause of urethritis, prostatitis and epididymitis in men. In the male urogenital tract, the bacteria or soluble products of their metabolism may interact with sperm, and alter their quality. AIMS: To determine the in vitro effect of incubation of N. gonorrhoeae and soluble products of their metabolism on sperm quality and assess the bacteria-sperm interaction. METHODS: Soluble products of bacterial metabolism and 0.5 McFarland concentration of N. gonorrhoeae were incubated with semen samples from healthy volunteers. Conventional (motility and viability) and functional sperm parameters (potential of mitochondrial membrane integrity and sperm membrane lipid peroxidation, detection of reactive oxygen species, chromatin integrity and expression of annexin V) were quantified using microscopy and cytometry flow, respectively. To assess the interaction of N. gonorrhoeae to human sperm microscopic observation was performed. RESULTS: N. gonorrhoeae can join the sperm and decreases sperm viability after 1.5 hours of incubation with human sperm (84.5% vs 66.5%, p<0.05), without affecting the functional sperm parameters. CONCLUSION: N. gonorrhoeae interacts with human sperm affecting sperm viability.

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.

Infecciones bacterianas del tracto reproductivo masculino y su papel en la fertilidad / Bacterial infections of the male reproductive tract and their role in fertility

Antecedentes: El espermatozoide es una célula altamente especializada encargada de llevar el material genético paterno hasta el tracto reproductivo femenino en búsqueda del oocito, no obstante durante su desplazamiento puede interactuar con sustancias, otras células e incluso microorganismos que puede transportar desencadenando procesos infecciosos que alteran el éxito reproductivo. Objetivo: Describir las bacterias involucradas en la alteración de la función reproductiva y sus efectos sobre calidad espermática, así como la capacidad de los espermatozoides para transportar infecciones y diseminarlas al tracto reproductivo femenino. Resultados: Las infecciones bacterianas que afectan el tracto reproductivo masculino se clasifican en infecciones de transmisión sexual, del tracto urinario y las asociadas a la microbiota bacteriana, donde están implicadas una gran variedad de agentes etiológicos como Chlamydia trachomatis, Ureoplasma urealyticum, Mycoplasma hominis, Escherichia coli y los Staphyloccoccus coagulasa negativos. Aún es controversial el efecto de estos gérmenes sobre los parámetros seminales así como la presencia de microbiota en el semen; su diagnóstico depende de la calidad de la muestra, de la sensibilidad de la técnica de detección y de los factores de riesgo que presente el individuo. Conclusión: Los procesos inflamatorios e infecciosos en el trato reproductivo masculino influyen en la fertilidad, por lo que se requiere profundizar en el estudio de estos procesos, establecer más y mejores métodos diagnósticos y pautas para el autocuidado que disminuyan la propagación de estos agentes patógenos.

Introduction: Spermatozoon is a highly specialized cell responsible for carrying the paternal genetic material toward the oocyte. During its journey in the female reproductive tract, the sperm cell interacts with substances, cells and also microorganisms that can be transported, thus triggering infectious processes that could alter reproductive success. Objective: Describe the species of bacteria involved in the alteration of sperm cells reproductive function and their effect on the sperm quality. In addition, evaluate the ability of the sperm cell to carry infections and disseminate them in the female reproductive tract. Results: Bacterial infections that affect the male reproductive tract are classified as sexually transmitted infections, urinary tract infections, and infections associated with bacterial microbiome. Chlamydia trachomatis, Ureoplasma urealyticum, Mycoplasma hominis, Escherichia coli and coagulase-negative Staphylococcus are the main etiologic agents of these infections. The presence of bacteria in semen and their effect of in semen parameters are still controversial. To probe that bacteria are present in semen depends on the quality of the sample, the sensitivity and specificity of the detection technique and the patient risk factors. Conclusion: Inflammatory and infectious processes in the male reproductive treatment affect fertility; therefore, further study of the role of thoseprocesses, establishment of more and better diagnostic methods and guidelines for self-care to decrease the spread of these pathogens will be necessary.

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.

Detection of Mycoplasma DNA from the sperm specimens of infertile men by PCR

Infections in accessory sex glands are considered as potential hazards to male fertility. These infections can affect different sites of the male reproductive tract such as the testis, epididymis and male accessory sex glands. Transmission of these infections to female partners causes genital infection, infertility and abortion. The aim of this study was to detect Mycoplasma, as one of the causatives, from the semen of infertile males using PCR assay. The survey considered 100 infertile men who referred to clinics and had not used antibiotics for 7 days prior to sampling. The infertility of cases was confirmed by a physician specialist. The sperm specimens were collected in sterile condition and sent to the laboratory rapidly. Specimens were examined for presence of Ureaplasma urealiticum and Mycoplasma hominis by PCR. Meanwhile, the history of vaginal infections and abortion in the female sexual partners was investigated. The results of multiplex PCR were compared with spermogram. All patients had no symptoms of genital infection. Of 100 infertile men, 33 [33%] were positive for CMU organisms [Chlamydia, Mycoplasma and Unreaplasma]. Uureaplasma urealiticum and Mycoplasma hominis were detected in 17 and 3 of patients respectively. Due to some problems in culturing of CMU organisms, PCR can be used as a diagnostic technique to detect such pathogens from seminal fluid of infertile men that leads to choose appropriate therapy in a shortest time

Genital trct infection in asymptomatic infertile men and its effect on semen quality

Male urogenital tract infection plays an important role in men infertility. Asymptomatic bacteriospermia has been paid attention as a major cause of male infertility. The aim of this study was to microbiological investigation of semen sample of infertile men attending to infertility clinic and evaluation of the effects of bacteriospermia on semen quality. Eighty eight infertile men were evaluated by standard bacterial culture method. Standard semen analysis was performed according to WHO guidelines. Among total cases, 35.22% [31 cases] showed at least one pathogen: 10.22% E.coli, 9.09% Coagulase Negative Staphylococci [Saprophyticcus], 6.81% Group B Streptococci, 5.88% Entrococci, 5.68% Candida sp., 2.27% Gonococci, 2.27% Staphylococcus aureus, 1.13% Klebsiella sp. and 1.13% Providencia sp. There was a significant relation between the bacteriospermia and the rate of no motile and morphologically abnormal sperms [P<0.0001]. The quality of sperm motility was significantly decreased in contaminated semen. The percentage of morphologically normal sperm was lower. E.coli and Entrococci were the most effective agents against sperm parameters. Asymptomatic bacteriospermia has a negative effect on sperm quality. E.coli and Entrococci are the most common bacteria with negative influence on sperm motility and morphology. Moreover, presence of bacteriospermia and leukocytospermia did not correlate with each other [P>0.05]. It seems that leukocytospermia is a poor marker to predict bacteriospermia

Correlation between asymptomatic urethritis with bacteriospermia in seminal plasma of fertile and infertile men

Urogenital infection can influence the fertilization potential of the spermatozoa, which may lead to male infertility. The symptomatic urethiritis can change the semen parameters, however the role of asymptomatic urethirtis in fertility is still obscure. In this descriptive investigation, a total of 148 samples from urethra and semen of fertile as well as 146 samples from urethra and semen of infertiles men were examined for the presence of 5 bacterial species including streptococcus group A., Entercoccus, E.coli, coagulase positive and negative Staphylococcus. The rate of infection of urethra and semen of fertile men were 49.22% and 29.05%, respectively. The aforementioned rates were 34.9% and 60.27% for infertile men. The seminal infection was significantly different between two groups of fertile and infertile [p < 0.01]. The results showed that the most common pathogen in semen of fertile and infertile men was Entrococcus with prevalence of 32.60% [14 cases], and 42% [37 cases], respectively. A total of 90 out of 131 samples contaminated with bacteria showed high rate of Leuckocytes [pyospermia]. The remaining 41 infected samples lacked or had low number of leuckocytes. In conclusion, bacteriospermia is significantly higher in seminal samples of infertile than fertile men [p < 0.01]. Therefore, the pathogens involved in urethritis may be involved in male infertility. In addition, seminal culture is necessary for detection of bacteria presence in the semen, and thus it is important to note that presence or absence of Leukocyte in semen may not represent the urogenital infection

Improvement of semen quality in infected asymptomatic infertile male after bacteriological cure

Bacteriological etiology was investigated in 29 infected asymptomatic infertile males. The localization of the infection and the effect of a long term antibiotic therapy on semen parameters were evaluated. The most frequent etiological agent isolated was Enterococcus faecalis. Positive bacteriological culture was obtained in prostatic fluid in 16 patients and in semen in 13. Bacteriological cure was achieved in 24 cases and it was associated with improved seminal parameters: sperm concentration, motility, viability and total motile sperm per ejaculate. In 5 patients without bacteriological cure there was no change in semen analysis after antibiotic therapy. In 45 percent of the infected patients there were less than 0.5 x 10(6)/ml seminal polymorphonuclear leukocytes. In view of these findings granulocyte concentration seems to be a poor marker to predict infection.

Prueba poscoital: correlación de la misma con los resultados del cultivo del moco cervical, del espermocultivo, del cultivo del líquido prostático y del espermograma / Postcoital test: its correlation with results from the culture of cervical mucus, spermoculture, culture of prostatic fluid and spermogram

Se hace un recuento histórico donde se pone de manifiesto el origen de la prueba poscoital o de Sims-Hünner, así como la importancia que el mismo ha tenido y aún tiene en el estudio de la pareja infértil. Se realiza un estudio de los resultados de los test poscoitales realizados a 50 parejas. Se correlacionan dichos resultados con los obtenidos del cultivo del moco cervical, del espermocultivo, del cultivo del líquido prostático y del espermograma

Transporte de la Escherichia coli por el espermatozoide humano a través del moco cervical mesocíclico / Escherichia coli transport by the human spermatozoa through the mesocyclic cervical mucous

Se sugiere que los Espermios humanos podrían actuar como vehículos de transporte de microorganismos: en nuestro trabajo hemos observado la unión específica de E. Coli a los mismos. La penetración de los espermios depende del estado del moco cervical durante el ciclo menstrual: Es máxima en el moco de tipo E (Estrogénico) y ausente en el moco de tipo G (Progestativo). Estudiamos la migración de los espermios a través del moco cervical Mesocíclico "In Vitro". La Microfotografía electrónica de barrido nos permitió observar una estrecha relación entre bacterias y espermios humanos. Nuestros resultados permiten apoyar la hipótesis de que este mecanismo mediado por espermios podría explicar el origen de procesos inflamatorios pelvianos