Hematogenous dissemination of Chlamydia muridarum from the urethra in macrophages causes testicular infection and sperm DNA damage†.

The incidence of Chlamydia infection, in both females and males, is increasing worldwide. Male infections have been associated clinically with urethritis, epididymitis, and orchitis, believed to be caused by ascending infection, although the impact of infection on male fertility remains controversial. Using a mouse model of male chlamydial infection, we show that all the major testicular cell populations, germ cells, Sertoli cells, Leydig cells, and testicular macrophages can be productively infected. Furthermore, sperm isolated from vas deferens of infected mice also had increased levels of DNA damage as early as 4 weeks post-infection. Bilateral vasectomy, prior to infection, did not affect the chlamydial load recovered from testes at 2, 4, and 8 weeks post-infection, and Chlamydia-infected macrophages were detectable in blood and the testes as soon as 3 days post-infection. Partial depletion of macrophages with clodronate liposomes significantly reduced the testicular chlamydial burden, consistent with a hematogenous route of infection, with Chlamydia transported to the testes in infected macrophages. These data suggest that macrophages serve as Trojan horses, transporting Chlamydia from the penile urethra to the testes within 3 days of infection, bypassing the entire male reproductive tract. In the testes, infected macrophages likely transfer infection to Leydig, Sertoli, and germ cells, causing sperm DNA damage and impaired spermatogenesis.

Human Papillomavirus and Infertility.

Background and objectives. Human papillomavirus (HPV) is the most commonly sexually transmitted infection. Recent evidence suggests that an HPV infection may affect fertility. The aim of the study was to determine the prevalence of HPV infections among couples undergoing in vitro fertilization (IVF) and to identify their awareness of HPV. Material and Methods. A total of 200 samples were collected from couples who received IVF treatment during 2017-2018 in Vilnius University Hospital Santaros Klinikos (VUH SK) Santaros Fertility Centre (SFC). For HPV detection, cervical swabs from women and sperm samples from men were taken and a real time polymerase chain reaction (RT-PCR) was used for the identification of 14 high-risk HPV types. Sperm parameters were evaluated according to World Health Organization (WHO) recommendations for 2010. Research subjects answered an anonymous questionnaire to ascertain their knowledge of HPV. Results. After testing of HPV in couples undergoing IVF, it was found that 33 out of 100 couples (33%) were HPV positive. Of these, 19% of women (19/100) and 20% of men (20/100) tested positive. Using Fisher's exact test, a statistically significant difference was found between HPV infections and abnormal sperm quality parameters (p = 0.023). Conclusions. HPV may have an impact in spermatogenesis, because an HPV infection was more frequently detected in men with abnormal sperm parameters. High-risk HPV 52 was the most common genotype among couples undergoing IVF treatment.

Carcinogenic Activities and Sperm Abnormalities of Methicillin Resistance Staphylococcus aureus and Inhibition of Their Virulence Potentials by Ayamycin.

This investigation aimed to study the in vivo harmful effects of the subcutaneous injection of different methicillin resistance Staphylococcus aureus extracts (MRSA2, MRSA4, MRSA10, MRSA69, MRSA70, MRSA76, and MRSA78). Such strains represented the highest minimum inhibition concentration toward methicillin with various multidrug-resistant patterns. The obtained results revealed that rats injected with the MRSA4 extract died immediately after the last dose indicating the high cytotoxicity of MRSA4 strain (100% mortality). While the mortalities in other groups injected by the other MRSA extracts ranged from 50 to 75%. In comparison with the normal animal group, all MRSA extracts induced a hepatotoxic effect which was indicated from the significant (p < 0.01) increases in the activities of the serum alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) enzymes. Moreover, alkaline phosphatase (ALP) combined with a partial nephrotoxicity that was monitored from the significant elevation of serum urea concentration. While serum creatinine levels did not affect. Similarly, a significant elevation was recorded in serum levels of tumor biomarkers (alpha fetoprotein; AFP, carcinoembryonic antigen; CEA, and lactate dehydrogenase; LDH) reflecting their carcinogenic potential. On the other hand, the percentage of micronuclei (MN) in polychromatic erythrocytes from bone marrow cells was statistically significant in all groups as compared to the control group. The percentage of sperm abnormalities was statistically significant compared to the control. Different types of head abnormalities and coiled tail were recorded. Consequently, the current study focused on fighting MRSA virulence factors by the new compound ayamycin, which proved to be potent anti-virulence factor against all MRSA strains under study by significant decreasing of their streptokinase activities, hemolysin synthesis, biofilm formation, and their cell surface hydrophobicity.

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.

Effect of human papillomavirus and Chlamydia trachomatis co-infection on sperm quality in young heterosexual men with chronic prostatitis-related symptoms.

OBJECTIVE: To investigate the effect of human papillomavirus (HPV) and Chlamydia trachomatis (Ct) co-infection on sperm concentration, motility and morphology, in a large cohort of young heterosexual male patients with chronic prostatitis-related symptoms. PATIENTS AND METHODS: Patients with chronic prostatitis-related symptoms, attending the same centre for sexually transmitted diseases from January 2005 and December 2010, were consecutively enrolled in this cross-sectional study. All patients underwent clinical and instrumental examination, microbiological cultures for common bacteria, DNA extraction, mucosal and serum antibodies evaluation for Ct, specific tests for HPV and semen analysis. The semen variables analysed were: volume; pH; sperm concentration; motility; and morphology. Subjects were subdivided in two groups: group A, patients with Ct infection alone and group B, patients with Ct and HPV co-infection. The main outcome measurement was the effect of Ct and HPV co-infection on the semen variables examined. RESULTS: Of 3050 screened patients, 1003 were enrolled (32.9%) in the study. A total of 716 (71.3%) patients were allocated to group A, and 287 (28.7%) to group B. Significant differences between the two groups were reported in terms of percentage of motile sperm (degrees of freedom [df] = 1001; t-test = 11.85; P < 0.001) and percentage of normal morphological forms (df = 1001; t-test = 7.18; P < 0.001), while no differences were reported in terms of semen volume or pH. According to World Health Organization thresholds for normal semen, 364 (50.8%) men in group A and 192 (66.8%) men in group B were subfertile (odds ratio = 1.95; 95% confidence interval 1.46-2.60; P < 0.001). No correlation between HPV genotype, mucosal IgA type and semen variables was found. CONCLUSION: In a population of prostatitis-related symptoms attributable to Ct infection, co-infection with HPV has a significant role in decreasing male fertility, in particular with regard to sperm motility and morphology.

[Generalized mycoplasma infection in patients and carriers].

AIM: Study of possibility of generalization of mycoplasma infection in patients with urogenital pathology. MATERIALS AND METHODS: Among the examined patients 5 males characterized by risky sexual behavior with pronounced symptoms of infection or without those were selected. Patients were examined by a complex of methods for the presence of mycoplasma infection by culture, PCR, DFA, PHA, AHR and by detection of specific immune complexes in blood sera. Scrapes from urogenital tract, blood sera samples, urine, saliva, prostatic fluid were materials for the study. RESULTS: In blood of all patients in ELISA antibodies against Mycoplasma hominis were detected; in PHA they were detected only in 2 individuals. In all the patients in blood CIC were detected including antigens and DNA of one or several mycoplasma species. Sperm of 3 individuals was infected by Ureaplasma spp., 2--M. genitalium. In saliva of 2 individuals M. hominis was detected, 3--U. urealyticum. CONCLUSION: In all the examined patients the infection was shown to have generalized character. This phenomenon presents itself as quite significant because mycoplasma may cause anti-apoptotic and oncogenic effect.

Influence of epididymitis on reproductive function.

In our study 54 patients were diagnosed epididymitis caused by STI and in 6 (11.1%) cases inflammatory process spread on seminal vesicles which negatively affected the state of male reproductive function. After having unilateral epididymitis pathospermia developed in 76% of cases and infertility - in 55.6%. In patients who had unilateral acute epididymitis, infertility caused by urethritis of chlamidial aetiology developed in 70.4% of cases, infertility caused by urethritis of mycoplasmatic (ureaplasma) genesis - in 62.9% of cases and infertility caused by gonorrheal urethritis - in 29.6% of cases. At this moment following alterations of ejaculate are revealed: volume reduction, decrease in spermatozoid number, reduction of ability to move, increase in teratomatous forms, spermagglutination, etc. The best way for prevention of pathospermia and infertility is using modern and adequate treatment methods directed to maximally effective suppression of acute inflammatory process in epididymis and then restoration of its functions.

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.

Effects of Ureaplasma urealyticum infection on the male reproductive system in experimental rats.

To study the effects of Ureaplasma urealyticum (Uu) infection on the male reproductive system, the mechanism of infertility induced by Uu infection was investigated in experimental rats. Male Sprague-Dowley rats were infected with Uu4 (serotype 4) through repeated natural sexual intercourse for 8 weeks to establish infection. After 8 weeks, the blood samples of the animals were collected and analysed for cytokine production, and the animals were microdissected for the analysis of the reproductive system. Morphological study showed that spermatozoa exhibited curling and breaks in the rats infected at different dosages. Of the infected rats, 27.5% had both soft and hard calculi in the urinary tract, compared with 12% in the control groups. Uu infection resulted in a decline of sperm quality, eventually leading to the death of the spermatozoa. In the infected animals, the serum interleukin 6 and interleukin 8 levels increased significantly (P < 0.05), while tumour necrosis factor-alpha and interferon-gamma showed only modest changes. Our observations showed that Uu infection has an impact on sperm morphology, leading to the death of the spermatozoa. It is plausible that the morphological alterations of spermatozoa induced by Uu infection are one of the possible factors that contribute to male infertility.

Expression of IL-6, IL-8, TNF-alpha, IL-10, HSP-60, anti-HSP-60 antibodies, and anti-sperm antibodies, in semen of men with leukocytes and/or bacteria.

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.

Spermatozoa and seminal plasma induce a greater inflammatory response in the ovine uterus at oestrus than dioestrus.

Leukocyte infiltration and increased synthesis of cytokines in response to insemination is considered to enhance reproductive success. The present study investigated the inflammatory response to whole semen, spermatozoa and seminal plasma, with and without the addition of antibiotics, in the ovine uterus at oestrus and dioestrus. Seminal plasma and spermatozoa both contributed to increased IL-8 secretion (P < 0.01) by endometrial epithelial cells and a concurrent infiltration by neutrophils (P < 0.01). Increased GM-CSF secretion (P < 0.01) occurred in response to whole semen and spermatozoa when antibiotics were not used. Macrophages and eosinophils increased (P < 0.05) in the endometrial stroma when antibiotics were not used, and fewer mast cells were detected in the deep endometrial stroma after treatments containing antibiotics (P < 0.05). Neutrophil and IL-8 responses to insemination were greater at oestrus (P < 0.01) than at dioestrus and the GM-CSF response followed a similar trend. Eosinophil numbers were increased at oestrus (P < 0.01) but minimally affected by insemination. More macrophages were located in the superficial endometrial stroma at oestrus. These results indicate that spermatozoa, seminal plasma and possibly bacteria contribute to the post-insemination inflammatory response, and that leukocytes, GM-CSF and IL-8 secretion in the ovine uterus are influenced by ovarian hormones.

Wolbachia modification of sperm does not always require residence within developing sperm.

Wolbachia are maternally inherited intracellular bacteria known to manipulate the reproduction of their arthropod hosts. Wolbachia commonly affect the sperm of infected arthropods. Wolbachia-modified sperm cannot successfully fertilize unless the female is infected with the same Wolbachia type. A study of spermatogenesis in the parasitic wasp Nasonia vitripennis reveals that Wolbachia are not required in individual spermatocytes or spermatids to modify sperm. In N. vitripennis, Wolbachia modify nearly all sperm, but are found only in approximately 28% of developing sperm, and are also found in surrounding cyst and sheath cells. In the beetle Chelymorpha alternans, Wolbachia can modify up to 90% of sperm, but were never observed within the developing sperm or within the surrounding cyst cells; they were abundant within the outer testis sheath. We conclude that the residence within a developing sperm is not a prerequisite for Wolbachia-induced sperm modification, suggesting that Wolbachia modification of sperm may occur across multiple tissue membranes or act upstream of spermiogenesis.

Ultrastructure of reproductive system of female Litomosoides chagasfilhoi Moraes-Neto, Lanfredi and De Souza, 1997 (Nematoda: Filarioidea).

The ultrastructure of the female reproductive system of the filariid Litomosoides chagasfilhoi by transmission electron microscopy (TEM) is described for the first time. The ovary is composed of primary oocytes surrounded by a single layer of epithelial cells apposed on the basal laminae. The ovarian wall is completely filled with primary oocytes, which are arranged radially and are centrally connected around the rachis. The uterine wall consists of muscular fibers surrounded by a basal lamina and the epithelium underlying the lamina. Ameboid and aflagellate spermatozoa are present inside the distal portion of the uterus, some of them near oocytes, which present bacteria in its cytoplasm. An eletrondense well-defined eggshell covers the zygotes, which presents in its cytoplasm bacteria arranged in groups. These bacteria are also observed in embryos and in the hypodermal cord. These ultrastructural aspects of L. chagasfilhoi female worms presented herein contribute to the knowledge of the morphology and embryonary development of this filariid, providing means for further comparative analyses of the action of anti-filarial drugs. Besides this, the presence of bacteria Wolbachia-like is being reported for the first time in this species, showing the great importance of this experimental model of study.

Effects of Wolbachia on sperm maturation and architecture in Drosophila simulans Riverside.

Wolbachia is an intracellular obligate symbiont, that is relatively common in insects and also found in some nematodes. Cytoplasmic incompatibility (CI) is the most commonly expressed form, of several sex altering phenotypes caused by this rickettsial-like bacterium. CI is induced when infected males mate with uninfected females, and is likely the result of bacterial-induced modification of sperm grown in a Wolbachia-infected environment. Several studies have explored the dynamics of Wolbachia bacteria during sperm development in Drosophila. This study confirms and extends these earlier investigations of Wolbachia's distribution and proliferation in male germ cell lines. We examined Wolbachia population dynamics during testis development of Drosophila simulans (Riverside) by studying their distribution during the early mitotic divisions of secondary spermatogonial and subsequent meiotic cyst cells. Wolbachia are found in lower concentration in spermatogonial than in spermatocyte cells. Cytoplasmically incompatible crosses result in low levels of viable embryos despite the occurrence of fairly high levels of uninfected cysts. During meiotic divisions Wolbachia organize themselves at the poles during prophase and telophase but arrange themselves in equatorial bands during metaphase and anaphase. Moreover, during meiosis Wolbachia are asymmetrically divided between some daughter cells. There is no strong relationship between the fusome and Wolbachia and we have not found evidence that bacteria cross the ring canals. Wolbachia were observed at the distal and proximal sides of individualization complexes. Multiple altered sperm structures were observed during the process of individualization of infected sperm.

Prevalence of sexually transmissible pathogens in semen from asymptomatic male infertility patients with and without leukocytospermia.

OBJECTIVE: To determine the prevalence of pathogens that cause sexually transmitted infections (STIs) in semen from asymptomatic male infertility patients with and without leukocytospermia (LCS), and associations between STIs, inflammatory markers, and other semen variables. DESIGN: Retrospective, controlled study. SETTING: Academic Medical Center. PATIENT(S): Two hundred and forty-one male infertility patients undergoing routine semen analysis: 132 with LCS, and 109 without LCS. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The DNA from STI pathogens (human papillomavirus [HPV], cytomegalovirus [CMV], herpes simplex virus [HSV], human herpesvirus type 6 [HHV-6], Epstein-Barr virus [EBV], hepatitis B virus [HBV], and Chlamydia trachomatis [CT]), routine semen parameters, and markers of accessory gland and epididymal function and inflammation. RESULT(S): The DNA from STI pathogens was detected in 45/241 (18.7%) of the samples (CMV, 8.7%; HPV, 4.5%; HHV-6, 3.7%; HSV, 3.7%; CT, 2.5%; EBV, 0.4%; and HBV, 0%), with no difference in prevalence between the LCS and non-LCS groups. The DNA of STI pathogens in semen was associated with a decrease in sperm concentration, motile sperm concentration, total sperm count, and neutral alpha-glucosidase concentration, whereas LCS was associated with a decrease in total sperm count, percent normal forms, and fructose concentration. CONCLUSION(S): The DNA of STI pathogens was detected in semen from a high percentage of asymptomatic male infertility patients, and was associated with poor semen quality. Efforts to diagnose and treat subclinical genital-tract infections should be intensified.