Bacteria In Semen Research Articles

Reliable Detection of Excessive Sperm Ros Production in Subfertile Patients: How Many Men with Oxidative Stress?

Sperm oxidative stress has been extensively associated to male infertility. However, tests to detect this parameter have not been yet introduced in clinical practice and no definitive data are present on the extent of oxidative stress in male infertility. In this study, we used a novel and reliable flow cytometric method to reveal sperm ROS production in subfertile patients (n = 131) and in healthy donors (n = 31). Oxidative stress was higher in subfertile patients (14.22 [10.21-22.08]%) than in healthy donors (9.75 [8.00-14.90]% (p < 0.01)), but no correlation was found with age, semen quality or sDF. We also failed to detect an increase in sperm ROS production with semen viscosity or leukocytospermia, but a sharp impact of semen bacteria was evident (with bacteria: 31.61 [14.08-46.78]% vs. without bacteria: 14.20 [10.12-22.00]%, p < 0.01). Finally, after establishing a threshold as the 95th percentile in healthy donors, we found that 29% of subfertile patients exceeded this threshold. The percentage decreased to 25.56% when we excluded subjects with bacteriospermia and increased to 60.87% when only these patients were considered. In conclusion, 29% of subfertile patients showed an excessive sperm ROS production. Surprisingly, this parameter appears to be independent from routine semen analysis and even sDF determination, promising to provide additional information on male infertility.

The Interaction between Canine Semen Bacteria and Semen Quality Parameters.

Assessing canine semen quality helps to detect infertility in males, but identifying factors that influence canine semen quality is a complicated task. The objective of this study was the assessment of the potential influence of bacteria found in canine semen samples on the characteristics of dogs' semen. In this study, semen samples were collected manually from 30 dogs and subjected to a comprehensive examination. The results of sperm motility, concentration, viability, and morphology were statistically analysed in relation to the number of bacteria in the semen (CFUs/mL) and the seminal microbiota. Samples with an increased bacterial count per millilitre were associated with lower-quality sperm motility (p < 0.05). The most frequently isolated bacterial genera from the analysed semen samples were Staphylococcus spp. (26.0%), Corynebacterium spp. (17.8%), and Streptococcus spp. (16.4%). The presence of β-haemolytic Escherichia coli bacteria was linked to suboptimal semen samples, characterised by significantly reduced semen viability and a lower proportion of morphologically normal spermatozoa (p < 0.05). Corynebacterium spp. was associated with reduced bacterial load and superior semen quality (p < 0.01). These findings highlight the importance of bacterial cell counts and microbiota diversity in relation to various factors influencing canine semen quality, providing a more comprehensive understanding of canine reproductive well-being.

The Impact of Microorganisms on Canine Semen Quality.

Various microorganisms, including Mycoplasma spp., have been reported in canine ejaculate. The impact of these microorganisms on semen quality remains unclear. This study included 63 male intact healthy dogs aged 1-8 years. One dog exhibited azoospermia, indicating a relatively low incidence of this condition. Interestingly, 36.5% of the examined dogs tested negative for both aerobic bacteria and mycoplasmas, while 12.7% tested positive for bacterial presence. Additionally, 60.3% of the dogs tested positive for Mycoplasma spp. using PCR, with most carrying 1-2 Mycoplasma species. We found no significant difference in semen characteristics between Mycoplasma-positive and -negative dogs. The detection of Mycoplasma was not significantly linked to the presence of bacteria in semen. All the microorganisms identified were classified as saprophytic flora. Our findings indicate that Mycoplasma spp. is common in canine ejaculate. Semen quality parameters were not correlated with the presence of Mycoplasma spp. in semen. Mycoplasma HRC689 was the most common species. Some dogs exhibited no presence of aerobic bacteria or mycoplasmas in their semen. Our study highlights the common presence of Mycoplasma spp. in canine ejaculate. Semen quality shows no correlation with Mycoplasma presence. Some canine ejaculate is sterile. Our findings suggest the existence of undescribed species of canine mycoplasmas, necessitating advanced diagnostic techniques like NGS for their identification.

Ejaculatory Abstinence Affects the Sperm Quality in Normozoospermic Men-How Does the Seminal Bacteriome Respond?

This study was designed to describe bacterial profiles of ejaculates collected following a long and short ejaculatory abstinence set in the context of changes in the conventional, oxidative, and immunological characteristics of semen. Two specimens were collected in succession from normozoospermic men (n = 51) following 2 days and 2 h, respectively. Semen samples were processed and analyzed according to the World Health Organization (WHO) 2021 guidelines. Afterwards, sperm DNA fragmentation, mitochondrial function, levels of reactive oxygen species (ROS), total antioxidant capacity, and oxidative damage to sperm lipids and proteins were evaluated in each specimen. Selected cytokine levels were quantified using the ELISA method. Bacterial identification by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that samples collected following two days of abstinence presented with a higher bacterial load and diversity, and a greater prevalence of potentially uropathogenic bacteria including Escherichia coli, Staphylococcus aureus and Enterococcus faecalis. Only staphylococci and Escherichia coli remained present in specimens obtained after 2 h of abstinence. Whilst all samples accomplished the criteria set by WHO, a significantly higher motility (p < 0.05), membrane integrity (p < 0.05), mitochondrial membrane potential (p < 0.05), and DNA integrity (p < 0.0001) were detected following 2 h of ejaculatory abstinence. On the other hand, significantly higher ROS levels (p < 0.001), protein oxidation (p < 0.001), and lipid peroxidation (p < 0.01) accompanied by significantly higher concentrations of tumor necrosis factor alpha (p < 0.05), interleukin-6 (p < 0.01), and interferon gamma (p < 0.05) were observed in specimens collected after two days of abstinence. It may be summarized that shorter ejaculatory abstinence does not compromise sperm quality in normozoospermic men, while it contributes to a decreased occurrence of bacteria in semen which is accompanied by a lower probability of damage to spermatozoa by ROS or pro-inflammatory cytokines.

Pilot Study: Next-generation Sequencing of the Semen Microbiome in Vasectomized Versus Nonvasectomized Men

BackgroundApproximately half a million vasectomies are performed every year in the USA. There is a paucity of literature on the impact of male sterilization on the semen microbiome and whether it prompts microbiota dysbiosis. ObjectiveTo investigate if vasectomy induces changes in the seminal microbiome via comparison of semen samples from men before and after vasectomy, and if the seminal microbiome profiles for vasectomized men follow a particular pattern with respect to diversity and abundance. Design, setting, and participantsFrom July 2021 to February 2022, we prospectively collected and analyzed semen samples from 58 men at one outpatient clinic. Eighteen men provided a semen sample before and 3 mo after vasectomy. We also collected semen samples from 22 fertile nonvasectomized men and from a further 18 vasectomized men at 3 mo after vasectomy. Outcome measurements and statistical analysisSemen microbiome α-diversity, beta-diversity, and relative abundance were compared initially between paired and then between unpaired vasectomized and nonvasectomized samples. Analysis of variance (ANOVA), permutational multivariate ANOVA, and analysis of the composition of microbiomes with bias correction were used to assess differences. Results and limitationsIn both paired and unpaired sets of samples, a decreasing trend for α-diversity in semen after vasectomy was observed. Shannon diversity, the relative abundance of species with an abundance >2%, and composition were not significantly changed. Sphingomonas, Brevundimonas, and Paracoccus abundance decreased after vasectomy, while Corynebacterium abundance increased. The results may be limited by the sample size and lack of demographic heterogeneity. ConclusionsVasectomy is followed by a decrease in α-diversity and changes in the relative abundance of bacterial species in the semen microbiome. Further investigation is necessary to understand the clinical significance of these changes after vasectomy. Patient summaryWe evaluated changes in the bacteria species in semen after vasectomy. We found that vasectomy decreased the richness and evenness of bacteria species in semen, but the overall bacterial community remained similar. Further studies are needed to assess the implications of changes in semen bacteria after vasectomy.

Bacteriospermia - A formidable player in male subfertility.

Bacterial colonization of male reproductive tissues, cells, and fluids, and the subsequent impact of bacteria on the sperm architecture, activity, and fertilizing potential, has recently gained increased attention from the medical and scientific community. Current evidence strongly emphasizes the fact that the presence of bacteria in semen may have dire consequences on the resulting male fertility. Nevertheless, the molecular basis underlying bacteriospermia-associated suboptimal semen quality is sophisticated, multifactorial, and still needs further understanding. Bacterial adhesion and subsequent sperm agglutination and immobilization represent the most direct pathway of sperm–bacterial interactions. Furthermore, the release of bacterial toxins and leukocytic infiltration, associated with a massive outburst of reactive oxygen species, have been repeatedly associated with sperm dysfunction in bacteria-infested semen. This review serves as a summary of the present knowledge on bacteriospermia-associated male subfertility. Furthermore, we strived to outline the currently available methods for assessing bacterial profiles in semen and to outline the most promising strategies for the prevention and/or management of bacteriospermia in practice.

The microbiome of seminal fluid

Manole COJOCARU Department of Physiology and Immunology, “Titu Maiorescu” University, Faculty of Medicine, Bucharest, Romania Abstract The microbiome of semen is a field of increasing scientific interest. Compared to other body sites, the seminal microbiota has been minimally investigated. Recent studies indicate that presence of bacteria in semen is relatively frequent, including in fertile individuals

Alteration of Some Sperm Proteins, Anti-oxidants, and Prostate-specific Antigen in the Seminal Plasma of Bacteriospermic Infertile Males

Aim: To investigate the alteration of some sperm proteins, anti-oxidants, and prostate-specific antigen in the seminal plasma of bacteriospermic infertile males.&#x0D; Study Design: The study is a case-control design to investigate the semen parameters and some sperm proteins in infertile males with bacteriospermia attending urology or fertility clinics in Port Harcourt, Rivers State, Nigeria.&#x0D; Place and Duration of Study: The study was carried out in the Department of Medical Laboratory Science, Rivers State University, Port Harcourt. However, some of the Laboratory investigations were done in the Chemical Pathology Unit and Medical Microbiology Unit of the University of Port Harcourt Teaching Hospital, Port Harcourt. The Study was between the period of September 2019 and Feb. 2022.&#x0D; Methodology: A total 193 subjects were recruited of which 76 were fertile without bacterial infection, 24 were fertile with bacterial infection, 37 infertile without bacterial infection and 56 infertile with bacterial infections. Infertile males were further classified as teratozoospermic, asthenoteratozoospermic, and oligoasthenoteratozoospermic. HSP70, PKA, MDA, TAC, and GPX (in seminal plasma), PSA, and testosterone (in serum) were analyzed using ELISA while TAC was done using spectrophotometric methods. Microbial culture was done to isolate and identify bacteria in semen. Results obtained were analyzed using GraphPad Prism and SPSS.&#x0D; Results: Culture results indicated that 52% of normospermic subjects had no semen infection at the time of the investigation. On the other hand, infertile male subjects without semen infections had 18.7%, while infertile males with semen infections were seen to be 29.2%. Likewise, subjects with bacteriospermia as scanty growth had 12.5%, 9.3% moderate, and 7.3% heavy growth. Significantly higher values of HSP70, PKA, PSA, and MDA were seen in infertile males with bacteriospermia compared with control and infertile males without infection while significantly lower values were seen in TAC, GPX, OPN, and testosterone were observed in the infertile males with bacteriospermia compared with control and infertile males without infection.&#x0D; Conclusion: Bacteriospermia significantly increased lipid peroxidation cum oxidative stress leading to altered physiological sperm proteins in the seminal plasma. These alterations could also be responsible for the poor sperm qualities observed in the infertile subjects.

Icariin improves pig sperm quality through antioxidant and antibacterial effects during liquid storage at 17 °C

Oxidative stress and bacterial contamination are two main factors affecting pig sperm quality. The objective of this study was to evaluate the effects of Icariin (ICA) on sperm quality, antioxidative and antibacterial activity during liquid storage at 17 °C. Boar semen was diluted into Beltsville thawing solution (BTS) containing different concentrations of ICA (0, 5, 10, 15, 20 and 25 mg/L). During storage, total sperm motility, progressive motility, kinematic parameters, acrosome and plasma membrane integrity, total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity, malondialdehyde (MDA) content and reactive oxygen species (ROS) levels, bacterial content and the substitution effect of ICA for gentamicin (GEN) were assessed on days 1, 3 and 5, respectively. The results showed that supplementation of ICA in the extender not only significantly improved the antioxidant properties of sperm but also inhibited the growth of bacteria in semen, with 15 mg/L ICA showing the best performance on sperm quality. Furthermore, co-treatment of 15 mg/L ICA with 62.5 mg/L GEN showed similar effects on sperm quality and bacterial inhibition as 250 mg/L GEN, indicating that 15 mg/L ICA can replace 75% of the 250 mg/L GEN dose in semen extenders. Overall, we found that ICA can exert both antioxidant and antimicrobial effects to improve sperm quality and can also be served as an antibiotic substitute.

Semen microbiota and cytokines of healthy and infertile men.

In this study, we determined the levels of cytokine secretory inhibitors and the microbiota biofilms of semen from healthy and infertile subjects. A total of 118 clinical bacterial isolates were isolated and tested. Cytokine secretory inhibitors were determined based on the difference in cytokine content between the control and experimental samples of cell-free supernatants of isolated microorganisms. Biofilm formation was studied by determining the adhesion of microorganisms to the surface of a 96-well sterile plate and expressed as the optical density at 630 nm (OD630). Cell-free supernatants of Staphylococcus contained higher levels of secretory inhibitor of cytokines in conditionally healthy than in infertile patients. In contrast, in infertile men, the ability to reduce cytokine levels was more characteristic of Enterococcus and Corynebacterium. Seminal Staphylococcus, Corynebacterium, and Enterococcus isolated from infertile subjects showed a greater ability to form biofilms than the same bacteria isolated from healthy men. Further research is needed on this topic, since it is necessary to determine the relationships between decreased secretory inhibitors of cytokines, production of biofilms by bacteria in semen, and infertility. It is likely that the ability of microorganisms to change the concentration of cytokines and increase the level of biofilm formation in semen may be associated with minimal impairments of fertilizing ability, which are not detected using other methods.

Antimicrobial Resistance in Equine Reproduction.

Simple SummaryBacteria can develop resistance to antibiotics, resulting in the appearance of infections that are difficult or impossible to treat. This ability enables bacteria to survive in hostile environments and can result from exposure to even small amounts of antibiotic substances. Bacteria are present in the reproductive tract of the horse; they can develop resistance to antibiotics, because the animal has been treated for an infection, or due to insemination with a semen dose that contains antibiotics. Bacteria colonize the membrane lining the male reproductive tract and are transferred to the semen during collection. They can cause sperm quality to deteriorate during storage or may cause an infection in the mare. Therefore, antibiotics are added to the semen dose, according to legislation. However, these antibiotics may contribute to the development of resistance. Current recommendations are that antibiotics should only be used to treat bacterial infections and where the sensitivity of the bacterium to the antibiotic has first been established. Therefore, adding antibiotics to semen extenders does not fit these recommendations. In this review, we examine the effects of bacteria in semen and in the inseminated mare, and possible alternatives to their use.Bacteria develop resistance to antibiotics following low-level “background” exposure to antimicrobial agents as well as from exposure at therapeutic levels during treatment for bacterial infections. In this review, we look specifically at antimicrobial resistance (AMR) in the equine reproductive tract and its possible origin, focusing particularly on antibiotics in semen extenders used in preparing semen doses for artificial insemination. Our review of the literature indicated that AMR in the equine uterus and vagina were reported worldwide in the last 20 years, in locations as diverse as Europe, India, and the United States. Bacteria colonizing the mucosa of the reproductive tract are transferred to semen during collection; further contamination of the semen may occur during processing, despite strict attention to hygiene at critical control points. These bacteria compete with spermatozoa for nutrients in the semen extender, producing metabolic byproducts and toxins that have a detrimental effect on sperm quality. Potential pathogens such as Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa may occasionally cause fertility issues in inseminated mares. Antibiotics are added during semen processing, according to legislation, to impede the growth of these microorganisms but may have a detrimental effect on sperm quality, depending on the antimicrobial agent and concentration used. However, this addition of antibiotics is counter to current recommendations on the prudent use of antibiotics, which recommend that antibiotics should be used only for therapeutic purposes and after establishing bacterial sensitivity. There is some evidence of resistance among bacteria found in semen samples. Potential alternatives to the addition of antibiotics are considered, especially physical removal separation of spermatozoa from bacteria. Suggestions for further research with colloid centrifugation are provided.

Low density Porcicoll separates spermatozoa from bacteria and retains sperm quality

Antibiotics are added to semen extenders to control the growth of bacteria contaminating semen during collection but may contribute to the development of antibiotic resistance. An alternative would be physical separation of spermatozoa from bacteria. The objective of the present study was to evaluate two low densities of Porcicoll for removal of bacteria, and for their effect on sperm recovery and sperm quality. Semen was collected from boars at a commercial station. Aliquots of 8 extended ejaculates were subjected to colloid centrifugation through 20% Porcicoll (P20) and 30% Porcicoll (P30) in 500 mL tubes and then stored at 17 °C. Microbiological examination and sperm quality evaluation (computer assisted sperm analysis and flow cytometry) were carried out on controls and all colloid-selected samples immediately after preparation and again after storage for 3 and 7 days. The microorganisms found were mainly bacteria from the environment, gut or skin. There was a considerable reduction or complete removal of some bacteria by both colloids. Recovery rates were 86% for P20 and 81% for P30. Sperm quality was not adversely affected by colloid centrifugation on day 0, and thereafter showed a more gradual deterioration in colloid centrifuged samples than in controls, possibly due to lower bacterial contamination. There were no differences in sperm quality between the two colloid treatments. Thus, these results show that contaminating bacteria in semen can be controlled by centrifugation through low density colloids.

Bacterial Analysis for Seminal Fluid befor In-vitro Fertilization Procedure

In vitro fertilization (IVF) is the most advanced technique of assisted reproductive where the sperm reach the eggs for fertilization in a laboratory tube. Seminal fluid often contains bacterial contaminants that decrease the quality and quantity of seminal fluids.&#x0D; Aim: In this study we investigated the presence of bacteria in semen before IVF procedure.&#x0D; Methodology: All samples were identified according to the morphological, biochemical, molecular, and antimicrobial tests.&#x0D; Results: Microbial species were detected in 28% of semen samples, while 72% did not show any bacterial growth. The prevalent bacterial isolates were Staphylococcus spp., Enterococcus spp., Catellicoccus spp., and Pseudomonas spp.&#x0D; Conclusion: The use of microbiological analysis to identify resistant bacterial isolates is considered an effective method to reduce infertility in men by changing the contaminated semen.

Effect of rosmarinic acid on bull sperm quality and bacteria in semen

Effect of rosmarinic acid on bull sperm quality and bacteria in semen

Metagenomic analysis of bacteria in stallion semen

Bacteria colonize stallion semen during collection and processing which may cause disease in inseminated females or negatively affect sperm quality during storage prior to insemination. Antibiotics are added to semen extenders to control the growth of these bacteria but may induce antimicrobial resistance. Research into alternatives to antibiotics for this purpose requires knowledge of which bacteria are present in semen. Not all bacteria in semen, however, can be identified by conventional microbiological techniques. The objectives of the study were to: i) determine which bacteria are present in stallion semen using metagenomics; and ii) investigate individual differences in bacterial content in semen from all stallions on one premises. Bacterial DNA was extracted from ejaculates from seven stallions (one ejaculate per stallion) and bacteria were identified using 16S sequencing. In total, 83 bacterial genera were identified, varying from 25 to 52 among different individuals. There was a negative correlation (r = -0.81212; P < 0.05) between the presence of Treponema spp. and Advenella spp. In conclusion, most of the bacteria present in stallion semen could be identified to genus level by 16S sequencing even when present at a low frequency. This method of identification may help to clarify individual variation in bacterial content and its potential effects on fertility.

The Prevalence of Bacteriospermia in Infertile Men and Association with Semen Quality in Southwestern Iran

Infertility considered as a social and public health issue and estimated that most of these infertile couples are residents of developing countries. Infectious diseases including the history of Sexually Transmitted Infections (STIs) may impact on male reproductive function. Therefore, the present study aimed to investigate the prevalence of bacterial contaminants of semen and probable association with sperm quality of infertile men in Iranian population. The study population consisted of 200 infertile men and 150 fertile men attending an infertility Center in southwestern Iran during the study period in 2015. The assessment of sperm parameters was according to the World Health Organization (WHO) guidelines. The presumptive pathogens were identified using standard microbiology tests and confirmed by specific PCR primers. The prevalence of bacteriospermia in the semen of the infertile group was significantly higher than that in the fertile group (48% vs. 26.7%, P <0.001). The microbiological analysis of samples showed that the most abundant species of bacteria in semen of infertile men were Chlamydia trachomatis (12.5%) followed by Neisseria gonorrhoeae (11%). On the other hand, in the control group, Lactobacillus spp. (17.3%) was the most isolated pathogen. Results showed that the presence of N. gonorrhoeae, C. trachomatis, Mycoplasma genitalium, Haemophilus, and Klebsiella was significantly associated with sperm abnormality. Based on our findings, it seems that bacteriospermia is associated with alterations in the properties of semen which may lead to a decrease in the fertilization potential of sperm. Therefore, immediate and appropriate treatment is necessary before investigating every other possible cause of infertility.

Quality of frozen semen of Brahman bulls used for routine artificial insemination in Bangladesh

Background: Semen must be of good quality to achieve satisfactory conception rate in any artificial insemination (AI) programme. The objectives of the present study were to evaluate the quality of frozen semen of Brahman bulls and compare the same semen derived from different sources. Methods: Fifteen frozen semen straws of Brahman bulls derived from 3 different sources (5 straws from each source) were evaluated with respect to volume, motility, concentration of spermatozoa and morphology of spermatozoa. Volume of semen was determined by micropipette, concentration was evaluated by haemocytometer technique, motility and progressive motility were evaluated by computer assisted sperm analyser (CASA), spermatozoa with normal acrosome, midpiece and tail were evaluated in formol-saline fixed semen and spermatozoa with normal head morphology were evaluated by Farley staining technique. Moreover, presence or absence of bacteria in semen was evaluated by Gram’s staining technique. Results: The overall semen volume, concentration, motile percentage, progressive motile percentage, percentage of spermatozoa with normal acrosome, midpiece and tail, and percentage of normal head morphology were 0.19±0.01 ml, 30.87±3.4x106/ml, 56.47±24.2 (%), 16.23±9.8 (%), 88.23±2.1 (%) and 95.1±1.6 (%), respectively. When compared, similar volume of semen (0.19 ml) was found in all frozen semen derived from different sources. The difference in percentage of motile spermatozoa between source B and C was significant (P&lt;0.05). The difference in spermatozoa with normal acrosome, midpiece and tail in semen between source A and B was significant (P&lt;0.05). The difference in the percentage of progressive motile spermatozoa, the concentration of spermatozoa and normal head morphology in semen among different sources of semen was not significant (P&gt;0.05). The difference in percentage of motile spermatozoa, the percentage of progressive motile spermatozoa, the concentration of spermatozoa, normal acrosome, midpiece and tail, normal head morphology in semen between locally produced and foreign semen was not significant (P&gt;0.05). All examined 15 semen samples were negative for presence of bacteria. Conclusions: Although the quality of frozen semen derived from Brahman bulls seems to be within normal limit, the post-AI conception rate should be determined to evaluate the fertilizing capacity of the used semen.

How does the microbial load affect the quality of equine cool-stored semen?

Contaminating bacteria present in stallion ejaculates may compromise sperm quality during storage. Different procedures have been used to reduce the load of microorganisms in semen and avoid bacterial growth during storage. The aims of this study were: 1) to evaluate different techniques to eliminate bacteria in semen 2) to study the relationship between total microflora load (TML) and ROS production; and 3) to determine if TML affects the functionality of cool-stored sperm. Ejaculates from 11 stallions were split and processed in 3 ways: A. extended semen; B. conventional centrifuged semen, and C. Single layer centrifugation through Androcoll-E (SLC). All samples were preserved in INRA 96 at 5 °C for 72 h. Aliquots from native semen and from different treatments were taken for bacteriological analysis at T0, T24, T48 and T72h of storage and Total microbial load (TML: CFU (colony-forming units/ml) was calculated. The ROS production (dichlorodihydrofluorescein diacetate for H2O2, dihydroethidium for superoxide anion and CellROX deep red for total ROS), viability (YO-PRO-1-Ethidium) and lipid peroxidation (BODIPY-C11) were assessed by flow cytometry, and motility by CASA. The bacteria isolated were Corynebacterium spp, Arcanobacterium spp, Bacillus spp, Dermobacter, Staphylococcus spp, Streptococcus spp, Penicilium spp. TML of semen showed correlations with live sperm (r: −0.771), dead sperm (r: 0.580), H2O2 production (r: 0.740), and total ROS production (CellROX (+)) (r: −0.607), Total motility (r: 0.587), Progressive motility (r: −0.566), VCL (r: −0.664), VSL (r: −0,569), VAP (r: −0.534) (p ≤ 0.05). SLC removed 99.34% of the microbial load, which was assicated with a significanlty reduced H2O2 production (p ≤ 0.05). However, only samples treated with Androcoll-E had a higher total ROS production (CellROX +) (p ≤ 0.05). These results suggest that CellROX stain probably identifies superoxide production rather than H2O2 and this higher superoxide production may reflect an intense sperm functionality. The bacterial load increased the production of H2O2 in cool-stored semen which was associated with lower tolerance to refrigeration. SLC was the sperm processing technique that was most efficient at removing bacteria, reducing H2O2 production and selecting the most functional sperm.

Time trends for bacterial species and resistance patterns in semen in patients undergoing evaluation for male infertility

ABSTRACTSemen from asymptomatic men who are being evaluated as male partners in interfile couples have been reported to contain a variety of bacteria. Longitudinal studies of the variation of these bacteria over time and their resistance patterns have not been commonly reported. At our institution, residues from semen samples are routinely evaluated for bacteria, including antibiotic sensitivity profiles. We set out to profile the changes in semen bacteria and antibiotic resistance at our institution over time. A total of 72 semen isolates were examined for type of bacteria and sensitivity to a panel of antibiotics. The results were divided into two separate 5-year intervals (the first beginning in 2006, the second in 2011) and compared. The majority of bacteria were skin flora, with Streptococcus and Staphylococcus being the most prevalent. The resistance data for these two pathogens showed minimal statistically significant difference between the two time periods, although the Staphylococcus species did show a trend toward increasing resistance, suggesting that antibiotics currently used in sperm cell preparations may need to be varied.

Semen Bacterial Concentrations and HIV-1 RNA Shedding Among HIV-1-Seropositive Kenyan Men.

HIV-1 is transmitted through semen from men to their sexual partners. Genital infections can increase HIV-1 RNA shedding in semen, but shedding also occurs in the absence of typical pathogens. We hypothesized that higher bacterial concentrations in semen would be associated with higher HIV-1 RNA levels. We analyzed semen samples from 42 HIV-1-seropositive Kenyan men using quantitative polymerase chain reaction (PCR) to assess bacterial concentrations and real-time PCR to measure HIV-1 RNA levels. Generalized estimation equations were used to evaluate associations between these 2 measures. Broad-range 16S rRNA gene PCR with pyrosequencing was performed on a subset of 13 samples to assess bacterial community composition. Bacteria were detected in 96.6% of 88 samples by quantitative PCR. Semen bacterial concentration and HIV-1 RNA levels were correlated 0.30 (P = 0.01). The association between bacterial concentration and HIV-1 RNA detection was not significant after adjustment for antiretroviral therapy (ART) (adjusted odds ratio: 1.27, 95% CI: 0.84 to 1.91). Factors associated with semen bacterial concentration included insertive anal sex (adjusted beta 0.92, 95% CI: 0.12 to 1.73) and ART use (adjusted beta: -0.77, 95% CI: -1.50 to 0.04). Among 13 samples with pyrosequencing data, Corynebacterium spp., Staphylococcus spp., and Streptococcus spp. were most frequently detected. Most of these HIV-1-infected men had bacteria in their semen. ART use was associated with undetectable semen HIV-1 RNA and lower semen bacterial concentrations, whereas insertive anal sex was associated with higher bacterial concentrations. Additional studies evaluating the relationship between semen bacteria, inflammation, mucosal immunity, and HIV-1 shedding are needed to understand implications for HIV-1 transmission.