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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.

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OBJECTIVE: This study aimed to assess the levels of microbial contamination in semen samples before and after the micro swim-up (MSU) procedure in intra-cytoplasmic sperm injection (ICSI). The new method is an upgrade to the classic wash swim-up procedure. METHODS: Semen analysis and microbiological tests were carried out before and after the MSU procedure. A total of twenty semen samples were analyzed. RESULTS: Pathogens were observed in semen samples only before MSU and never after ICSI. Microbiological tests revealed a large prevalence of gram-positive cocci [Staphylococcus spp. (n=16, 80%) and viridans streptococci (n=10, 50%)]. The results of this study indicate that direct MSU in ICSI improved the ICSI workflow. CONCLUSION: The new workflow is faster and more affordable, and is likely to prevent infection problems that could arise from the normal microbial flora of the semen.

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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.

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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.

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OBJECTIVE: To determine the effect on human sperm of Escherichia coli strains separated on the basis of their ability to produce hemolysis. DESIGN: Experimental study. SETTING: University-based laboratory. PATIENT(S): Semen samples from healthy donors. INTERVENTION(S): Five million sperm, selected via the swim-up method, were incubated with 3 E. coli concentrations to obtain ratios of sperm to E. coli of 1:2, 1:16, and 1:128. The E. coli strains were: a hemolytic isolated strain (H), a nonhemolytic American Type Culture Collection strain (NH-ATCC), and a nonhemolytic isolated strain (NH-I). MAIN OUTCOME MEASURE(S): Aliquots of human sperm were used to measure progressive motility using computer-aided sperm analysis, mitochondrial membrane potential (ΔΨm) with a JC-1 (5,5',6,6' tetrachloro-1,1',3,3'-tetraethylbenzamidazolocarbocyanin iodide) and propidium iodide stain, and intracellular reactive oxygen species (iROS) with a dihydroethidium (DHE) stain. Sperm ΔΨm and iROS were measured by flow cytometry. Sperm vitality was considered the mean of propidium iodide-negative and DHE-negative cells. RESULT(S): Sperm incubated with the H strain in a 1:2 sperm to bacteria ratio demonstrated a significant decrease in motility and ΔΨm, and an increase of iROS. The NH-ATCC strain decreased sperm motility and ΔΨm, but in a ratio of sperm to bacteria of 1:128; it increased iROS at a ratio of 1:16. The NH-I strain did not affect the analyzed sperm functions, even at a 1:128 sperm to bacteria ratio. CONCLUSION(S): Results show a greater pathogenic effect on human sperm of E. coli strains with, versus without, hemolytic capacity.

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BACKGROUND: Cryopreservation and global trading of P. vannamei sperm will become a potential and important biotechnological tool. Nevertheless, information of the possible transfer of bacteria in cryopreserved shrimp sperm has not been registered yet. OBJECTIVE: The objective of this work was to determine the type of bacteria that could be cryopreserved together with white shrimp sperm masses. MATERIALS AND METHODS: Sixteen sperm masses were cryopreserved in 10% DMSO and 0.5 M trehalose and sixteen fresh sperm masses were used for bacterial analysis. Bacterial colonies were isolated and selected for sequencing. RESULTS: Strains were seawater borne and facultative aerobic bacteria from the genera Bacillus, Micrococcus, Paracoccus, Ruegeria and Staphylococcus. Most of them have been related with benefits to its host. None were pathogenic for P. vannamei. CONCLUSION: Cryopreservation implies preserving pathogenic or beneficial bacteria together with the sample. Therefore, it is possible to enhance cryopreserved samples or disperse pathogenic bacteria, which needs to be prevented.

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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.

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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.

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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.

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PROBLEM: Different cellular and biochemical markers have been proposed as indicators of infection-inflammation of male genital tract. METHOD OF STUDY: Semen samples from 80 men attending an andrologic clinic were evaluated to determine the presence of leukocyte, bacteria, antibodies against Chlamydia trachomatis, levels of IL-6, IL-8, IL-10, and TNF-alpha, HSP-60, anti-HSP-60 antibodies, and anti-sperm antibodies. RESULTS: Leukocytes in semen significantly correlated with an increase in IL-6, IL-8, and TNF-alpha. The simultaneous presence of pathogens and leukocytes was associated with high levels of IL-8 and TNF-alpha, whereas IL-6 was more associated with the presence of leukocytes. Anti-HSP-60 antibodies positively correlated with IL-6 and IL-8. The presence of anti-sperm antibodies highly associated with an increase in anti-HSP-60 antibodies. CONCLUSIONS: The type of cytokines present in the semen will depend on the single or simultaneous presence of leukocytes and/or pathogens. Chronic male genital tract infections could be associated with the development of anti-HSP-60 antibodies and anti-sperm antibodies.

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