[Is bacteriological screening of semen samples before in vitro fertilization treatment justified?] / Indokolt-e a spermaminták bakteriológiai szurése in vitro fertilizáció elott?

The number of couples seeking assisted reproductive technologies is increasing worldwide. The question of whether routine bacteriological screening of semen is necessary during the investigation and treatment of infertility is controversial. The semen sample often contains bacteria even if the hygiene rules for collection are followed. There is a growing number of studies dealing with the importance of the semen microbiome. Bacteriospermia can result not only from infection but also from contamination or colonization. Symptomatic infections or sexually transmitted diseases should be treated, but the relevance of asymptomatic positive cultures is controversial. Several studies have suggested that urinary tract infections may play a role in male infertility and that the quality of semen may be impaired by elevated bacterial or white blood cell counts. However, there are conflicting results on the effect of the treatment of bacteriospermia and leukocytospermia on sperm quality. Semen contaminated with microbes may also infect the embryos, thus compromising the success of treatment. In contrast, most studies have found no significant difference in the effectiveness of in vitro fertilization treatment in the presence or absence of bacteriospermia. This can be explained by the sperm preparation techniques, the antibiotic content of the culture media and the use of the intracytoplasmatic sperm injection technique. Thus, the need for routine semen culture before in vitro fertilization treatment and the management of asymptomatic bacteriospermia is questionable. Orv Hetil. 2023; 164(17): 660-666.

Penicillin and amikacin mixture has bactericidal activity equivalent to gentamicin, tylosin, lincomycin and spectinomycin mixture in equine frozen semen.

Neat stallion semen can contain a variety of microorganisms, some of which may impair sperm quality and/or cause infection of the mares' reproductive tract. For this reason, antibiotics are commonly added to semen extenders. A combination of gentamicin, tylosin, lincomycin and spectinomycin (GTLS) has been recommended for use, but there are no reports on the use of this mixture in equine semen extender. Penicillin and amikacin (PA) are safe for preserving sperm quality while effectively controlling bacterial growth in equine cooled stored semen, but data on frozen semen are scarce. Therefore, a bioequivalence study was performed to assess the bactericidal activity of GTLS and PA in equine frozen semen. Nine mature, healthy stallions were used in the study. Split ejaculates were processed using media without antibiotics (Control) or with different antibiotics. For the GTLS group, centrifugation medium and freezing extender were prepared with gentamicin 250 µg/ml, tylosin 50 µg/ml, lincomycin 150 µg/ml and spectinomycin 300 µg/ml. For the PA group, the centrifugation medium was prepared with potassium penicillin G (PPG) 1200 units/ml and the freezing extender was prepared with PPG 1200 units/ml and amikacin 500 µg/ml. Semen processed in extenders without antibiotics had higher (p < .005) bacterial loads throughout all cryopreservation processing steps than semen samples processed using antibiotics. There were no differences in semen bacterial load after centrifugation, 15 and 30 min after final extension, and after thawing between GTLS and PA groups, but PA had faster (p < .05) kill-time kinetics than GTLS. Only minor differences in sperm kinetic parameters were observed among groups. In conclusion, this study demonstrated bioequivalence between GTLS and PA in mitigating end-point bacterial loads. Prudent concentrations of the antibiotic mixtures evaluated in this study can be considered both effective and sperm-safe for equine frozen semen.

Altered circadian clock gene expression in the sperm of infertile men with asthenozoospermia.

PURPOSE: Male infertility is a complex multifactorial pathological condition, and asthenozoospermia (AZS) is one of the most common causes. Current evidence suggests the underlying role of the circadian clock on male fertility. This study aims to evaluate the expression levels of five principal clock genes in the sperm and their correlations with the sperm parameters in male infertility. METHODS: We determined the expression profiles of BMAL1, CLOCK, CRY1, PER1, and PER2 in the sperm of infertile men with AZS (n=38) and healthy fertile men (n=40) using quantitative real-time PCR. Then we performed comprehensive association analyses on the clock gene levels and the sperm parameters, including progressive and total motility, concentration, and normal morphology of the sperm. RESULTS: Our results showed that the expression levels of five clock genes (BMAL1, CLOCK, CRY1, PER1, and PER2) are significantly decreased in the sperm of the infertile men with AZS as compared with that of healthy fertile men (P< 0.01). All five clock gene levels are associated with the percentage of progressive/total sperm motility (r= 0.546/0.589~0.677/0.695, P< 0.01). We also discovered that a combination of BMAL1, CLOCK, CRY1, PER1, and PER2 could reach a high diagnostic performance (areas under the curves, 92%) for infertility with AZS. CONCLUSIONS: This study first reports that sperm BMAL1, CLOCK, CRY1, PER1, and PER2 levels are altered in AZS and may be molecular markers for male infertility with AZS. These findings indicate the possibility of stabilizing circadian rhythmicity through therapeutic intervention on clock genes to prevent and treat infertility.

Morphometric and morphokinetic differences in the sperm- and oocyte-originated pronuclei of male and female human zygotes: a time-lapse study.

PURPOSE: To study the morphometric and morphokinetic profiles of pronuclei (PN) between male and female human zygotes. METHOD(S): This retrospective cohort study included 94 consecutive autologous single day 5 transfer cycles leading to a singleton live birth. All oocytes were placed in the EmbryoScope + incubator post-sperm injection with all annotations performed retrospectively by one embryologist (L-SO). Timing parameters included 2nd polar body extrusion (tPB2), sperm-originated PN (tSPNa) or oocyte-originated PN (tOPNa) appearance, and PN fading (tPNF). Morphometrics were evaluated at 8 (stage 1), 4 (stage 2), and 0 h before PNF (stage 3), measuring PN area (um2), PN juxtaposition, and nucleolar precursor bodies (NPB) arrangement. RESULTS: Male zygotes had longer time intervals of tPB2_tSPNa than female zygotes (4.8 ± 0.2 vs 4.2 ± 0.1 h, OR = 1.442, 95% CI 1.009-2.061, p = 0.044). SPN increased in size from stage 1 through 2 to 3 (435.3 ± 7.2, 506.7 ± 8.0, and 556.3 ± 8.9 um2, p = 0.000) and OPN did similarly (399.0 ± 6.1, 464.3 ± 6.7, and 513.8 ± 6.5 um2, p = 0.000), with SPN being significantly larger than OPN at each stage (p < 0.05 respectively). More male than female zygotes reached central PN juxtaposition at stage 1 (76.7% vs 51.0%, p = 0.010), stage 2 (97.7% vs 86.3%, p = 0.048), and stage 3 (97.7% vs 86.3%, p = 0.048). More OPN showed aligned NPBs than in SPN at stage 1 only (44.7% vs 28.7%, p = 0.023). CONCLUSION(S): Embryos with different sexes display different morphokinetic and morphometric features at the zygotic stage. Embryo selection using such parameters may lead to unbalanced sex ratio in resulting offspring.

The influence of the female reproductive tract and sperm features on the design of microfluidic sperm-sorting devices.

Although medical advancements have successfully helped a lot of couples with their infertility by assisted reproductive technologies (ART), sperm selection, a crucial stage in ART, has remained challenging. Therefore, we aimed to investigate novel sperm separation methods, specifically microfluidic systems, as they do sperm selection based on sperm and/or the female reproductive tract (FRT) features without inflicting any damage to the selected sperm during the process. In this review, after an exhaustive studying of FRT features, which can implement by microfluidics devices, the focus was centered on sperm selection and investigation devices. During this study, we tried not to only point to the deficiencies of these systems, but to put forth suggestions for their improvement as well.

Total Aseptization of Boar Semen, to Increase the Biosecurity of Reproduction in Swine.

The aim of the study was to establish the complete microbiological profile of boar semen (Sus scrofa domesticus) and to choose the most effective antiseptic measures in order to control and optimize AI reproduction in pig farms. One hundred and one semen samples were collected and analyzed from several pig farms. The microbiological profile of ejaculates was determined by evaluating the degree of contamination of fresh semen and after dilution with specific extenders. The bacterial and fungal load of fresh boar semen recorded an average value of 82.41/0.149 × 103 CFU/mL, while after diluting the ejaculates the contamination value was 0.354/0.140 × 103 CFU/mL. Twenty-four germs (15 bacterial and 9 fungal species) were isolated, the most common being Candida parapsilosis/sake (92%) and Escherichia coli (81.2%). Modification of the sperm collection protocol (HPBC) reduced contamination in raw sperm by 49.85% in bacteria (significant (p < 0.00001) and by 9.67% in fungi (non-significant (p < 0.111491). The load in bacteria and filamentous fungi can be controllable, but not in levuras fungi. Some fluconazole-added extenders (12.5 mg%), ensure fungal aseptization, and even an increase in sperm progressivity (8.39%) for at least a 12 h shelf life after dilution. Validation of sperm aseptization was done by maintaining sow fecundity unchanged after AI (insignificant p > 0.05).

Assessing the testicular sperm microbiome: a low-biomass site with abundant contamination.

RESEARCH QUESTION: The semen harbours a diverse range of microorganisms. The origin of the seminal microbes, however, has not yet been established. Do testicular spermatozoa harbour microbes and could they potentially contribute to the seminal microbiome composition? DESIGN: The study included 24 samples, comprising a total of 307 testicular maturing spermatozoa. A high-throughput sequencing method targeting V3 and V4 regions of 16S rRNA gene was applied. A series of negative controls together with stringent in-silico decontamination methods were analysed. RESULTS: Between 50 and 70% of all the detected bacterial reads accounted for contamination in the testicular sperm samples. After stringent decontamination, Blautia (P = 0.04), Cellulosibacter (P = 0.02), Clostridium XIVa (P = 0.01), Clostridium XIVb (P = 0.04), Clostridium XVIII (P = 0.02), Collinsella (P = 0.005), Prevotella (P = 0.04), Prolixibacter (P = 0.02), Robinsoniella (P = 0.04), and Wandonia (P = 0.04) genera demonstrated statistically significant abundance among immature spermatozoa. CONCLUSIONS: Our results indicate that the human testicle harbours potential bacterial signature, though in a low-biomass, and could contribute to the seminal microbiome composition. Further, applying stringent decontamination methods is crucial for analysing microbiome in low-biomass site.

Effect of bacterial infection on sperm quality and DNA fragmentation in subfertile men with Leukocytospermia.

BACKGROUND: Although bacterial infections have been recognized as a possible cause of male infertility, the effect of bacterial infections on sperm quality and sperm DNA fragmentation remains controversial. The current study aimed to investigate the prevalence rate of bacterial infection in subfertile men and its effect on semen quality. Seminal fluid was collected from 172 male members of infertile couples attending the andrology infertility center and a group of 35 fertile subjects as a control. Sperm parameters and DNA fragmentation were evaluated based on the type of bacteria in all ejaculates. RESULTS: From the 172 patients investigated for infertility, 60 (34.88%) patients had a positive culture for pathogenic bacteria of different species. Leukocytospermia was significantly higher in infected samples in comparison with non-infected samples (p < 0.05). Sperm concentration and motility and morphology were significantly lower in infected than non-infected samples. Moreover, sperm DNA fragmentation was significantly higher in infected than non-infected samples. Besides, our results showed that sperm DNA fragmentation was correlated significantly with leukocytospermia (R: 0.22, p < 0.01). CONCLUSION: The present study suggested that bacterial infection significantly correlated with leukocytospermia could impair male fertility potential through decreasing sperm motility, morphology, and DNA integrity.

The impacts of cytoplasmic incompatibility factor (cifA and cifB) genetic variation on phenotypes.

Wolbachia are maternally transmitted, intracellular bacteria that can often selfishly spread through arthropod populations via cytoplasmic incompatibility (CI). CI manifests as embryonic death when males expressing prophage WO genes cifA and cifB mate with uninfected females or females harboring an incompatible Wolbachia strain. Females with a compatible cifA-expressing strain rescue CI. Thus, cif-mediated CI confers a relative fitness advantage to females transmitting Wolbachia. However, whether cif sequence variation underpins incompatibilities between Wolbachia strains and variation in CI penetrance remains unknown. Here, we engineer Drosophila melanogaster to transgenically express cognate and non-cognate cif homologs and assess their CI and rescue capability. Cognate expression revealed that cifA;B native to D. melanogaster causes strong CI, and cognate cifA;B homologs from two other Drosophila-associated Wolbachia cause weak transgenic CI, including the first demonstration of phylogenetic type 2 cifA;B CI. Intriguingly, non-cognate expression of cifA and cifB alleles from different strains revealed that cifA homologs generally contribute to strong transgenic CI and interchangeable rescue despite their evolutionary divergence, and cifB genetic divergence contributes to weak or no transgenic CI. Finally, we find that a type 1 cifA can rescue CI caused by a genetically divergent type 2 cifA;B in a manner consistent with unidirectional incompatibility. By genetically dissecting individual CI functions for type 1 and 2 cifA and cifB, this work illuminates new relationships between cif genotype and CI phenotype. We discuss the relevance of these findings to CI's genetic basis, phenotypic variation patterns, and mechanism.

Colloid centrifugation reduces bacterial load in chilled dog semen.

Conventional semen extenders contain antibiotics to prevent bacterial growth. Finding alternatives would be beneficial to minimize the development of bacterial resistance mechanisms. The aim of this study was to determine the effect of Single Layer Centrifugation (SLC) with Canicoll of dog semen on microbial load and sperm quality during cooled storage. Twenty-four ejaculates were obtained from healthy dogs by digital manipulation. Samples were diluted in Tris-citrate-fructose extender without antibiotics and divided into two treatment groups: SLC-selected samples and unselected samples. Sperm motility (CASA), viability and acrosome integrity (PI/FITC-PNA) as well as bacterial load of each microorganism species (colony-forming units/mL) were assessed at 0 and 48 h of storage at 4 °C. Results indicate SLC-selected dog spermatozoa have greater percentages of motility, viability and acrosome integrity (P < 0.05). Bacterial growth in SLC sperm samples was less (P < 0.05) than unselected samples. Removal of individual bacterial species varied from 91 % to 98 % for Escherichia coli (91.62 %), Streptococcus spp. (98.18 %), Staphylococcus spp.(95.33 %) and Pseudomonas spp. (92.50 %). In conclusion, the use of SLC with Canicoll has the potential to decrease bacterial load in chilled dog semen.

Effects of bacteria on male fertility: Spermatogenesis and sperm function.

Bacterial infection can negatively affect different parts of the male genital tract and subsequently cause impaired spermatogenesis and male fertility. However, most of the previous studies have focused on the infected organs of the male genital tract and there are not many studies that investigated the direct effect of bacteria on sperm and their mechanism of action. Interestingly, bacteria can induce different damages on sperm cells such as DNA fragmentation, cell membrane peroxidation, and acrosome impairment. Such negative effects can be mediated by bacteria-secreted toxins and metabolites or by direct attachment of bacteria on the sperm cells and subsequent activation of signaling pathways related to oxidative stress, apoptosis, and inflammation. These bacteria-induced changes can impair semen parameters and subsequently cause infertility. Given the significant destructive effect of some bacteria on sperm function and male fertility, in this study, we reviewed the impact of male urogenital bacteria on spermatogenesis and sperm functions as well as the underlying mechanisms by which the bacteria can damage sperm.

What human sperm RNA-Seq tells us about the microbiome.

PURPOSE: The study was designed to assess the capacity of human sperm RNA-seq data to gauge the diversity of the associated microbiome within the ejaculate. METHODS: Semen samples were collected, and semen parameters evaluated at time of collection. Sperm RNA was isolated and subjected to RNA-seq. Microbial composition was determined by aligning sequencing reads not mapped to the human genome to the NCBI RefSeq bacterial, viral and archaeal genomes following RNA-Seq. Analysis of microbial assignments utilized phyloseq and vegan. RESULTS: Microbial composition within each sample was characterized as a function of microbial associated RNAs. Bacteria known to be associated with the male reproductive tract were present at similar levels in all samples representing 11 genera from four phyla with one exception, an outlier. Shannon diversity index (p < 0.001) and beta diversity (unweighted UniFrac distances, p = 9.99e-4; beta dispersion, p = 0.006) indicated the outlier was significantly different from all other samples. The outlier sample exhibited a dramatic increase in Streptococcus. Multiple testing indicated two operational taxonomic units, S. agalactiae and S. dysgalactiae (p = 0.009), were present. CONCLUSION: These results provide a first look at the microbiome as a component of human sperm RNA sequencing that has sufficient sensitivity to identify contamination or potential pathogenic bacterial colonization at least among the known contributors.

Early Life Stages of a Common Broadcast Spawning Coral Associate with Specific Bacterial Communities Despite Lack of Internalized Bacteria.

Coral-associated bacteria are critical for the well-being of their host and may play essential roles during ontogeny, as suggested by the vertical transmission of some bacteria in brooding corals. Bacterial acquisition patterns in broadcast spawners remain uncertain, as 16S rRNA gene metabarcoding of coral early life stages suggests the presence of bacterial communities, which have not been detected by microscopic examinations. Here, we combined 16S rRNA gene metabarcoding with fluorescence in situ hybridization (FISH) microscopy to analyze bacterial assemblages in Acropora tenuis egg-sperm bundles, embryos, and larvae following a spawning event. Metabarcoding results indicated that A. tenuis offspring ≤ 4-day-old were associated with diverse and dynamic bacterial microbiomes, dominated by Rhodobacteraceae, Alteromonadaceae, and Oceanospirillaceae. While FISH analyses confirmed the lack of internalized bacteria in A. tenuis offspring, metabarcoding showed that even the earliest life stages examined (egg-sperm bundles and two-cell stages) were associated with a diverse bacterial community, suggesting the bacteria were confined to the mucus layer. These results can be explained by vertical transmission of certain taxa (mainly Endozoicomonas) in the mucus surrounding the gametes within bundles, or by horizontal bacterial transmission through the release of bacteria by spawning adults into the water column.

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

INTRODUCTION: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Bacteriospermia, extended spectrum beta lactamase producing Gram-negative bacteria and other factors associated with male infertility in Mwanza, Tanzania: a need of diagnostic bacteriology for management of male infertility.

BACKGROUND: Infections caused by Extended spectrum beta lactamase (ESBL) producing bacterial are global challenge. There is limited information on the magnitude of bacteriospermia, ESBL producing Gram-negative bacteria (GNB) causing bacteriospermia and factors associated with male infertility. This study determined magnitude of bacteriospermia, ESBL-GNB and other factors association with infertility among presumptive infertile men in Mwanza, Tanzania. METHODS: A cross-sectional hospital-based study was conducted between May 2017 and July 2018 among 137 presumptive infertile men. Semen specimens were self-collected by masturbation into clean, sterile and none-spermicidal containers and processed following laboratory standard operating procedures (SOPs). Data analysis was done using STATA 13.0. RESULTS: Gram-negative bacteria were predominantly isolated (86.4%), of which 31.6% were ESBL producers. In a total 44 bacteria were isolated from semen culture. The blaCTX-M gene was detected in 75% of phenotypically confirmed ESBL producers. Infertility was independently found to be associated with abnormal spermatozoa morphology (OR (95%CI): 14.48(3.17-66.05)) and abnormal spermatozoa motility (OR (95%CI): 0.05(0.01-0.24)). However, neither bacteriospermia (OR (95%CI): 0.86(0.29-2.59)) nor ESBL bacteriospermia (OR (95%CI): 0.13(0.01-1.22)) was found to be associated with infertility. CONCLUSION: One third of bacteriospermia is due to ESBL-producers with history of antibiotic use being protective factor for infertility. Abnormal spermatozoa morphology and poor spermatozoa forward motility independently predicted infertility.

Antibiotics Versus Natural Biomolecules: The Case of In Vitro Induced Bacteriospermia by Enterococcus Faecalis in Rabbit Semen.

Male subfertility is a global issue in human reproduction as well as in animal reproduction. Bacterial infection and semen contamination are still widely overlooked. As the collection of ejaculates is not a sterile process, it is necessary to add antimicrobial agents to avoid a possible depreciation of semen samples. As traditionally used antibiotics have been questioned because of an ever-increasing bacterial resistance, natural bioactive molecules could offer an alternative because of their antibacterial and antioxidant properties. As such, we decided to compare the effects of selected natural biomolecules (resveratrol-RES, quercetin-QUE and curcumin-CUR) with routinely used antibiotics in animal biotechnologies (penicillin-PEN, gentamicin-GEN and kanamycin-KAN) on the rabbit sperm vitality in the presence of Enterococcus faecalis. Changes in the sperm structural integrity and functional activity were monitored at 0, 2, 4 and 6 h. Computer-assisted sperm analysis (CASA) was used for the assessment of spermatozoa motility. Production of reactive oxygen species (ROS) was evaluated using chemiluminiscence, while the mitochondrial membrane potential (ΔΨm) was examined using the JC-1 dye. Finally, the sperm chromatin dispersion (SCD) test was used to assess DNA fragmentation, and changes to the membrane integrity were evaluated with the help of annexin V/propidium iodide. The motility assessment revealed a significant sperm motility preservation following treatment with GEN (p < 0.001), followed by PEN and CUR (p < 0.01). QUE was the most capable substance to scavenge excessive ROS (p < 0.001) and to maintain ΔΨm (p < 0.01). The SCD assay revealed that the presence of bacteria and antibiotics significantly (p < 0.05) increased the DNA fragmentation. On the other hand, all bioactive compounds readily preserved the DNA integrity (p < 0.05). In contrast to the antibiotics, the natural biomolecules significantly maintained the sperm membrane integrity (p < 0.05). The microbiological analysis showed that GEN (p < 0.001), KAN (p < 0.001), PEN (p < 0.01) and CUR (p < 0.01) exhibited the strongest antibacterial activity against E. faecalis. In conclusion, all selected biomolecules provided protection to rabbit spermatozoa against deleterious changes to their structure and function as a result of Enterococcus faecalis contamination. Therefore, administration of RES, QUE and/or CUR to rabbit semen extenders in combination with a carefully selected antibacterial substance may be desirable.

Klebsiella pneumoniae and Streptococcus agalactiae: Passengers in the sperm travel. / Klebsiella pneumoniae y Streptococcus agalactiae: Pasajeros en el viaje del espermatozoide.

During their journey through the female reproductive tract to reach the oocyte in the ampulla of the fallopian tube, spermatozoa interact with substances and microorganisms that affect sperm quality, thus altering their fertilizing capacity. OBJECTIVES: To determine in vitro the effect of Streptococcus agalactiae, Klebsiella pneumoniae and their soluble factors on sperm parameters, and to evaluate the ability of human sperm to interact with and transport these bacteria. METHODS: The effects of S. agalactiae, K. pneumoniae and their soluble factors on the viability, sperm motility and functional sperm parameters were quantified. In addition, motile spermatozoa were incubated with decreasing concentrations of bacteria for one hour, washed and post-infection treatments were performed with trypsin and transport capacity was assessed by quantitative cultures. RESULTS: Incubation of spermatozoa with K. pneumoniae decreased progressive motility. The soluble factors of K. pneumoniae increased the number of necrotic spermatozoa and the soluble factors of S. agalactiae increased lipid peroxidation of the sperm membrane (p<0.05). A strong interaction between sperm and bacteria was observed in the transport assays even in washed trypsin-treated samples. CONCLUSION: Human spermatozoa act as vectors for infections, generating strong interactions with K. pneumoniae and S. agalactiae favoring their diffusion through the female reproductive tract. This interaction affects male fertility by altering progressive motility, increasing the number of necrotic cells and inducing apoptosis.

Los espermatozoides durante su recorrido por el tracto reproductivo femenino en busca del oocito interactúan con sustancias y microorganismos que afectan la calidad seminal alterando su capacidad fecundante.OBJETIVOS: Determinar el efecto in vitro de Streptococcus agalactiae, Klebsiella pneumoniae y sus factores evaluar la capacidad de los espermatozoides humanos para interactuar y transportar estas bacterias.MÉTODOS: Se cuantificó el efecto de la incubación de S. agalactiae, K. pneumoniae y sus factores solubles sobre la viabilidad, la movilidad y los parámetros espermáticos funcionales. Adicionalmente, espermatozoides móviles fueron incubados con concentraciones decrecientes de bacterias durante una hora, se realizaron lavados y tratamientos post infección con tripsina y se evaluó la capacidad de transporte mediante cultivos cuantitativos.RESULTADOS: La incubación de los espermatozoides con K. pneumoniae ATCC 1705 disminuyó la movilidad espermática tipo I. Los factores solubles de K. pneumoniae ATCC 700603 aumentaron el número de espermatozoides necróticos mientras que los factores solubles de S. agalactiae aumentaron la lipoperoxidación de la membrana espermática (p<0,05). En los ensayos de transporte se observó una interacción fuerte entre los espermatozoides y las bacterias, incluso en las muestras tratadas con tripsina y lavados.CONCLUSIÓN: Los espermatozoides humanos sirven como vectores de infecciones, generan interacciones fuertes con K. pneumoniae y S. agalactiae, favoreciendo su difusión por el tracto reproductivo femenino. Esta interacción afecta la fertilidad masculina alterando la movilidad progresiva, aumentando el número de células necróticas y la apoptosis espermática.

Klebsiella pneumoniae and streptococcus agalactiae: passengers in the sperm travel / Klebsiella pneumoniae y Streptococcus agalactiae: Pasajeros en el viaje del espermatozoide

During their journey through the female reproductive tract to reach the oocyte in the ampulla of the fallopian tube, spermatozoa interact with substances and microorganisms that affect sperm quality, thus altering their fertilizing capacity. Objectives: To determine in vitro the effect of Streptococcus agalactiae, Klebsiella pneumoniae and their soluble factors on sperm parameters, and to evaluate the ability of human sperm to interact with and transport these bacteria. Methods: The effects of S. agalactiae, K. pneumoniae and their soluble factors on the viability, sperm motility and functional sperm parameters were quantified. In addition, motile spermatozoa were incubated with decreasing concentrations of bacteria for one hour, washed and post-infection treatments were performed with trypsin and transport capacity was assessed by quantitative cultures. Results: Incubation of spermatozoa with K. pneumoniae decreased progressive motility. The soluble factors of K. pneumoniae increased the number of necrotic spermatozoa and the soluble factors of S. agalactiae increased lipid peroxidation of the sperm membrane (p < 0.05). A strong interaction between sperm and bacteria was observed in the transport assays even in washed trypsin-treated samples. Conclusion: Human spermatozoa act as vectors for infections, generating strong interactions with K. pneumoniae and S. agalactiae favoring their diffusion through the female reproductive tract. This interaction affects male fertility by altering progressive motility, increasing the number of necrotic cells and inducing apoptosis

Los espermatozoides durante su recorrido por el tracto reproductivo femenino en busca del oocito interactúan con sustancias y microorganismos que afectan la calidad seminal alterando su capacidad fecundante. Objetivos: Determinar el efecto in vitro de Streptococcus agalactiae, Klebsiella pneumoniae y sus factores evaluar la capacidad de los espermatozoides humanos para interactuar y transportar estas bacterias. Métodos: Se cuantificó el efecto de la incubación de S. agalactiae, K. pneumoniae y sus factores solubles sobre la viabilidad, la movilidad y los parámetros espermáticos funcionales. Adicionalmente, espermatozoides móviles fueron incubados con concentraciones decrecientes de bacterias durante una hora, se realizaron lavados y tratamientos post infección con tripsina y se evaluó la capacidad de transporte mediante cultivos cuantitativos. Resultados: La incubación de los espermatozoides con K. pneumoniae ATCC 1705 disminuyó la movilidad espermática tipo I. Los factores solubles de K. pneumoniae ATCC 700603 aumentaron el número de espermatozoides necróticos mientras que los factores solubles de S. agalactiae aumentaron la lipoperoxidación de la membrana espermática (p < 0,05). En los ensayos de transporte se observó una interacción fuerte entre los espermatozoides y las bacterias, incluso en las muestras tratadas con tripsina y lavados. Conclusión: Los espermatozoides humanos sirven como vectores de infecciones, generan interacciones fuertes con K. pneumoniae y S. agalactiae, favoreciendo su difusión por el tracto reproductivo femenino. Esta interacción afecta la fertilidad masculina alterando la movilidad progresiva, aumentando el número de células necróticas y la apoptosis espermática

Effects of kojic acid on boar sperm quality and anti-bacterial activity during liquid preservation at 17 C.

Bacteriospermia is a documented risk to sperm quality when boar semen is stored at 17 °C. The objective of this study was to evaluate the effects of kojic acid (KA) on sperm quality and anti-bacterial effect during liquid storage boar semen at 17 °C, as well as to explore sperm-oocyte binding and embryonic development in vitro. Boar semen was diluted with Beltsville thawing solution (BTS), and it contained KA at different concentrations (0, 0.02, 0.04, 0.06, 0.08, and 0.10 g/L). Bacterial concentrations and sperm quality parameters (motility, mitochondrial membrane potential, acrosome integrity, and plasma membrane integrity) were evaluated on each experimental day. Differences in microbial compositions were compared using 16S rDNA sequencing among the control group, 0.04 g/L KA, and 0.25 g/L gentamycin groups on experimental day 5, and the effects of KA on sperm capacitation, Western blot, total anti-oxidant capacity (T-AOC), reactive oxygen species (ROS) content, malondialdehyde (MDA) content, in vitro fertilization (IVF) parameters, sperm-oocyte binding, cleavage rates, and blastocyst rates were evaluated. The results showed that KA at the optimum concentration of 0.04 g/L significantly improved sperm quality parameters and sperm capacitation, increased T-AOC ability, enhanced IVF parameters and sperm-oocyte binding, increased cleavage and blastocyst rates, inhibited bacterial concentrations, reduced ROS and MDA content, and altered bacterial compositions (P < 0.05). Moreover, KA also increased the expression of anti-oxidant-related proteins, SOD1, SOD2 and CAT, and anti-apoptosis-related protein, Bcl 2, and decreased the expression of apoptosis-related proteins, caspase 3 and Bax in sperm (P < 0.05). These findings demonstrated that supplementation of antibiotic-free extenders for boar semen with 0.04 g/L KA has beneficial effects on liquid boar sperm preservation.

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.