Genital Pathogens Research Articles

Performance verification and clinical evaluation of the NAP-Fluo Cycler system for detecting five genital tract pathogens based on microfluidic technology

IntroductionSexually transmitted infections (STIs) are among the most common infectious diseases worldwide, often leading to coinfections. Timely detection of genital tract pathogens in at-risk populations is crucial for preventing STIs. We evaluated the NAP-Fluo Cycler System, an innovative microfluidic nucleic acid detection platform, for its ability to simultaneously identify Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Ureaplasma urealyticum (UU), Mycoplasma genitalium (MG), and Mycoplasma hominis (MH) in urethral or cervical secretions. Materials and methodsThe limits of detection (LODs), repeatability, specificity, and interference resistance of the system were evaluated using standard strains, a panel of 24 pathogens, and seven interferents. We used the system to analyze 302 clinical samples and compared the results with those of five approved commercial reference kits. ResultsThe system achieved LODs of 500 IFU/mL, 500 CFU/mL, and 500 CCU/mL for CT, NG, and UU/MG/MH, respectively, demonstrating high stability (coefficient of variation <1.1 %), specificity, and resistance to interference. Among 302 clinical samples, 237 tested positive with single, dual, and triple infection rates of 35.6 %, 16.2 %, and 3.0 %, respectively. The reference kits detected 138 positive samples. The concordance rates with commercial reference kits were 100 % for UU, NG, and MH; 94.85 % for CT; and 80.00 % for MG. ConclusionsThis system offers a streamlined, rapid, and multiplex detection method that reduces testing time and complexity. Although it performs well with pure strains, it has limitations when using clinical samples of CT and MG, suggesting the need for further refinement before its widespread use in the clinic.

Impact of Genital Infections and Antibiotic Use on Incidence of Preterm Birth: A Retrospective Observational Study.

This study investigates the complex interplay among genital infections, antibiotic usage, and preterm birth. This study aims to identify common genital pathogens associated with preterm births, assess the impact of various antibiotic treatments on pregnancy outcomes, and understand antibiotic resistance patterns among these pathogens. This study included 71 pregnant women who experienced preterm birth and 94 women with genital infections who delivered at term. Various maternal characteristics, medical history, signs and symptoms, gestational weight, gestational age, type of birth, vaginal pH, Nugent scores, and vaginal flora were analyzed. Antibiotic resistance patterns of isolated microorganisms were also examined. The prevalence of sexually transmitted diseases (STDs) and genital herpes was significantly higher in the preterm group. Preterm births were associated with fever, pelvic pain, vaginal spotting, and fatigue. Vaginal pH levels and Nugent scores were significantly higher in the preterm group, indicating disturbed vaginal flora. The presence of Extended-Spectrum Beta-Lactamases (ESBLs) was a particularly strong risk factor, increasing by more than four times the odds of preterm birth (OR = 4.45, p = 0.001). Vancomycin-Resistant Enterococci (VRE) presence was another critical factor, with a four-fold increase in the odds of preterm birth (OR = 4.01, p = 0.034). The overall presence of Multidrug-Resistant (MDR) organisms significantly increased the odds of preterm birth (OR = 3.73, p = 0.001). Specific pathogens like Chlamydia trachomatis (OR = 3.12, p = 0.020) and Mycoplasma hominis (OR = 3.64, p = 0.006) were also identified as significant risk factors. Ureaplasma urealyticum also showed a significantly higher risk of preterm birth (OR = 2.76, p = 0.009). This study highlights the importance of screening for and treating genital infections during pregnancy, especially STDs and genital herpes, as they can significantly increase the risk of preterm birth. Additionally, the presence of specific microorganisms and antibiotic resistance patterns plays an essential role in preterm birth risk. Early detection and targeted antibiotic treatment may help mitigate this risk and improve pregnancy outcomes.

Effect of environmental factors on seminal microbiome and impact on sperm quality.

This review provides a comprehensive overview of the existing research on the seminal microbiome and its association with male infertility, while also highlighting areas that warrant further investigation. A narrative review was conducted, encompassing all relevant studies published between 1980-2023 on the male reproductive tract microbiome in humans. This review considered studies utilizing culture-based, polymerase chain reaction (PCR)-based, and next-generation sequencing (NGS)-based methodologies to analyze the microbiome. Data extraction encompassed sample types (semen or testicular tissue), study designs, participant characteristics, employed techniques, and critical findings. We included 37 studies comprising 9,310 participants. Among these, 16 studies used culture-based methods, 16 utilized NGS, and five employed a combination of methods for microorganism identification. Notably, none of the studies assessed fungi or viruses. All NGS-based studies identified the presence of bacteria in all semen samples. Two notable characteristics of the seminal microbiome were observed: substantial variability in species composition among individuals and the formation of microbial communities with a dominant species. Studies examining the testicular microbiome revealed that the testicular compartment is not sterile. Interestingly, sexually active couples shared 56% of predominant genera, and among couples with positive cultures in both partners, 61% of them shared at least one genital pathogen. In couples with infertility of known causes, there was an overlap in bacterial composition between the seminal and vaginal microbiomes, featuring an increased prevalence of Staphylococcus and Streptococcus genera. Furthermore, the seminal microbiome had discernible effects on reproductive outcomes. However, bacteria in IVF culture media did not seem to impact pregnancy rates. Existing literature underscores that various genera of bacteria colonize the male reproductive tract. These organisms do not exist independently; instead, they play a pivotal role in regulating functions and maintaining hemostasis. Future research should prioritize longitudinal and prospective studies and investigations into the influence of infertility causes and commonly prescribed medication to enhance our understanding of the seminal microbiota's role in reproductive health.

Escherichia coli and HPV16 coinfection may contribute to the development of cervical cancer

ABSTRACT Persistent human papillomavirus HPV infection is a necessary but insufficient condition for cervical cancer. Microorganisms are crucial environmental factors in cancers susceptibility and progression, recently attracting considerable attention. This study aimed to determine the infection status and relationship between high-risk HPV (HR-HPV) and lower genital tract infectious pathogens in cervical cancer and its precursors. From a retrospective and a prospective cohort analysis, Escherichia coli (E. coli) dominated the pathogens isolated from cervical discharges, and an isolation rate uptrend has been shown recently. HPV16 and E. coli’s coinfection rate gradually increased with the severity of cervical intraepithelial neoplasia. The adhesion and invasion abilities of the isolated E. coli to HPV16-positive SiHa cells were evaluated in vitro. The TCGA database and cervical tissues samples analysis showed that IL-10 was upregulated in cervical cancer. IL-10 expression levels increased in tissue samples with the severity of cervical cancer and its precursors with HPV16 and E. coli coinfection. Although no significant changes in IL-10 production were observed in the co-culture supernatant, we hypothesized that Treg immune cells in the tumour microenvironment might be responsible for the local IL-10 upregulation, according to our data showing Foxp3 upregulation and an upward trend with the cervical intraepithelial neoplasia grading to cancer and tumours with E. coli and HPV16 coinfection. Our data provide insights into the possible role of E. coli in cervical cancer progression and suggest that the application of HPV and E. coli screening programs may be an effective strategy to relieve the burden of cervical cancer and its precursor lesions.

Seroepidemiology of Treponema pallidum, Mycoplasma hominis, and Ureaplasma urealyticum in fertility treatment-seeking patients in the Emirate of Abu Dhabi, United Arab Emirates

BackgroundSeveral genital pathogens affect fertility. The study estimated the seroprevalence of Treponema pallidum, Ureaplasma urealyticum, and Mycoplasma hominis and identify specific factors associated with exposure to at least one of these pathogens in patients seeking fertility treatment in the Emirate of Abu Dhabi, United Arab Emirates. MethodsA seroepidemiological survey was conducted in a major fertility clinic in the Emirate of Abu Dhabi. Serum samples were screened for eight immunoglobulins (IgG, IgM, and IgA) against T. pallidum, U. urealyticum, and M. hominis using enzyme-linked immunoassays. Factors associated with seropositivity to at least one of the pathogens were investigated. ResultsThe study surveyed 308 patients seeking fertility treatment (mean age: 36.1 ± 6.8 years). Most patients were female (88.0%), 24.9% had at least one chronic comorbidity, 19.3% had a previous genital infection, and 68.1% had been diagnosed with infertility for ≥ 6 months. Ig seroprevalence of T. pallidum (IgG: 3.0%, IgM: 3.2%), U. urealyticum (IgG: 2.6%, IgM: 2.0%), and M. hominis (IgG: 33.9%) was 6.4%, 4.6%, and 49.0%, respectively. Nearly one quarter (23.0%) and one decile (9.2%) of the patients exhibited evidence of ongoing infection (IgM seropositivity) or recent infection (IgA seropositivity) with M. hominis, respectively. Overall, 53.0% of the patients were seropositive for at least one of the screened immunoglobulins. Patients with an education level of secondary schooling or below (66.2%) or those who were unemployed (61.1%) had a higher seroprevalence of IgG antibodies compared with patients with college or higher-level education (48.4%) or those who were employed (48.1%) (p < 0.05). ConclusionExposure to T. pallidum or U. urealyticum was relatively low, whereas that to M. hominis was common in the surveyed patients. Enhanced awareness and screening programmes for genital pathogens are crucial to prevent and control the transmission of infections and reduce the growing burden of infertility.

Global analysis on the mutations associated with multidrug-resistant urogenital mycoplasmas and ureaplasmas infection: a systematic review and meta-analysis

BackgroundThe emergence of multidrug-resistant (MDR) strains of genital pathogens, notably Mycoplasma genitalium and Ureaplasma spp., constitutes a significant global threat today. The present study aimed to evaluate the prevalence and trend of changes in MDR mycoplasma and ureaplasma strains.MethodsAn exhaustive search was performed across the ISI Web of Science, PubMed, Scopus, ScienceDirect, and Google Scholar databases to accumulate relevant studies without restrictions until April 2023. We used event rate and corresponding 95% confidence intervals to determine the frequency of resistance-related mutations and examine the trend of antibiotic resistance changes.ResultsThe data from 27 studies, including 24,662 patients across 14 countries, were evaluated. Out of the total studies, 20 focused on M. genitalium infections, and five on Ureaplasma spp. The frequency of resistance-associated mutations to macrolides, tetracyclines, and fluoroquinolones in clinical strains of M. genitalium was 43.5%, 13.1%, and 18.6%, respectively. The prevalence of M. genitalium strains with double resistance and MDR was 11.0% and 17.4%, respectively. The incidence of both double-drug-resistant and MDR strains was higher in the World Health Organization (WHO) Western Pacific Region than in European and American populations. For Ureaplasma strains, resistance-associated mutations to macrolides, tetracyclines, and fluoroquinolones were 40.8%, 25.7%, and 90.3%, respectively. The rate of antibiotic resistance was higher in the African population compared to the European and WHO Western Pacific Regions. The rate of MDR Ureaplasma infections was 13.2%, with a higher incidence in the African population compared to the WHO Western Pacific and European regions.ConclusionThe proliferation and spread of MDR Mycoplasma and Ureaplasma strains present a significant public health challenge. The situation is indeed alarming, and the rising trend of MDR M. genitalium and MDR Ureaplasma infections suggests that therapies involving macrolides and fluoroquinolones may become less effective.

The in vitro effects of probiotic bacteria on genital pathogens of female dogs

BackgroundAn important aspect in the microbiology of the reproductive system of small animals is the potential occurrence of probiotic bacteria, such as lactic acid bacteria (LAB) of the genus Lactobacillus. The presence of these microorganisms is significant due to their strong antibacterial and antifungal properties. This study aimed to select probiotic strains from the oral cavity and vagina that have outstanding antibacterial properties against typical genital pathogens of the female dog reproductive tract.ResultsThe antagonistic activity of ten LAB strains was tested against seven etiological agents isolated from the genital tract of female dogs with signs of inflammation. LAB strains with the greatest ability to inhibit the growth of indicator bacteria were Lactobacillus plantarum and L. acidophilus, while L. fermentum and L. brevis strains inhibited growth the least. Almost all strains showed a complete lack of adherence to Caco-2 epithelial cells.ConclusionsAll tested LAB isolates inhibited the in vitro growth of either Gram-positive or Gram-negative pathogens, suggesting that potential probiotic strains could contribute to the balance of the normal vaginal microbiota. Furthermore, they could be considered for use as prophylactic agents or as an alternative to antibiotic therapy for infections in dogs.

The association between adverse pregnancy outcomes and non-viral genital pathogens among women living in sub-Saharan Africa: a systematic review.

Adverse pregnancy outcomes are the main causes of maternal and neonatal morbidity and mortality, including long-term physical and psychological sequelae. These events are common in low- and middle-income countries, particularly in Sub Saharan Africa, despite national efforts. Maternal infections can cause complications at any stage of pregnancy and contribute to adverse outcomes. Among infections, those of the genital tract are a major public health concern worldwide, due to limited availability of prevention, diagnosis and treatment approaches. This applies even to treatable infections and holds true especially in Sub-Saharan Africa. As late as 2017, the region accounted for 40% of all reported treatable non-viral genital pathogens worldwide, many of which have been independently associated with various adverse pregnancy outcomes, and that include Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Treponema pallidum. Two databases (PubMed and Embase) were examined to identify eligible studies published up to October 2022. This study reviewed findings on the association between infections by treatable non-viral genital pathogens during pregnancy and adverse pregnancy outcomes among women living in Sub-Saharan Africa. Articles' title and abstract were screened at first using keywords as "sexually transmitted infections", "non-viral", "adverse pregnancy outcome", "Africa", "sub-Saharan Africa", "pregnant women", "pregnancy", and "pregnancy outcome". Subsequently, according to the eligibility criteria, potential articles were read in full. Results showed that higher risk of preterm birth is associated with Treponema pallidum, Chlamydia trachomatis and Candida albicans infections. Additionally, rates of stillbirth, neonatal death, low birth weight and intrauterine growth restriction are also associated with Treponema pallidum infection. A better insight on the burden of non-viral genital pathogens and their effect on pregnancy is needed to inform antenatal care guidelines and screening programs, to guide the development of innovative diagnostic tools and other strategies to minimize transmission, and to prevent short- and long-term complications for mothers and children.

Association of the uterine microbiome to mare fertility

Currently, limited information on the microbiome (community of microorganisms) and its role in infertility in the equine uterus is available beyond the detection of specific pathogens in some subfertile mares. Given that less than 1% of environmental bacteria are cultivable on common media (Handelsman, J. 2004, Microbiol Mol Biol Rev 68, 669-685.), it is likely that these cultured species represent a minority of the resident population. The overarching objectives of this project were to i) identify the resident uterine microbiome in estrus ii) assess how this microbiome evolves over time and iii) compare traditional culture-dependent methods of assessing the presence of bacteria in the equine uterus with 16S rRNA sequencing. Uterine cytobrush and swab samples were collected from 157 maiden Thoroughbred mares aged 8 years or younger and 116 Thoroughbred barren mares aged 4 to 15 years at the first reproductive exam of the breeding season. A second sample was collected from mares that were not pregnant at the end of the season (n=23). Furthermore, 20 of the mares were sampled in three consecutive breeding seasons. All cytobrush samples were placed and stored in RNAlater and kept at -20° C until further processing. Uterine swabs were plated on sheep blood and MacConkey plates, incubated at 37° C in aerobic conditions and assessed for bacterial growth at 24, 48 and 72 hours. DNA was extracted from cytobrush samples using routine methods. The genomic DNArecovered was used as a template for PCR amplification of the V3–V4 variable region of the bacterial 16S rRNA gene. Amplicons were sequenced on the Illumina MiSeq platform. DNA sequencing data was then processed to assess the equine microbiome composition. The richness (number of bacterial species) and evenness (distribution of bacterial numbers) of bacterial genetic sequences was compared between maiden, barren and wet mares and correlated with reproductive success. The microbiome was highly dynamic and changed significantly between seasons within mares with a limited “core microbiome” being detected. In some mares, conventional culture techniques failed to identify known equine genital pathogens that were detected using 16S rRNA sequencing. While the richness and evenness of the microbiome was comparable in maiden, dry and wet mares, the overall bacterial abundance was higher in mares that were barren at the beginning of the season and failed to become pregnant during the season, when compared to successfully bred barren mares and all maiden mares. This was the first time the microbiome of the equine uterus was studied over several timepoints/seasons. Future studies could explore a prognostic test that allows prediction of fertility in barren mares. This work was funded by AgriFutures Australia.

Isolation and characterization of bacteriophages specific to selected equine genital pathogens

Chronic bacterial endometritis is among the major problems in equine reproduction. Streptococcus equi ssp. zooepidemicus (SEZ), Klebsiella pneumoniae (KP), and Pseudomonas aeruginosa (PA) are among the most important pathogens for the condition. Bacteriophages (phages) are the viruses of bacteria, having a species- and strain-specific lytic activity. Due to their bactericidal capacities, phages can be used as an alternative treatment approach for bacterial infections. The study aimed to isolate and characterize phages with lytic activity against Streptococcus equi ssp. zooepidemicus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Phage samples were obtained from manure, uterine lavage fluid and drain water of horse husbandries (n=14). For isolation of phages, bacterial isolates of the equine genital tract (SEZ (n=1), KP (n=1), and PA (n=1)) were inoculated with the phage samples. For purification, bacteriophage plaques were repeatedly cultivated, harvested and filtered. For determining the host-range, the obtained phages were inoculated with bacterial isolates (SEZ (n=37), KP (n=26), and PA (n=33)). In total, 28 bacteriophages were isolated (against SEZ (n=13), KP (n=6), and PA (n=9)). Bacteriolysis was achieved in 32/37 Streptococcus-, 16/26 Klebsiella-, and 14/33 Pseudomonas-isolates. The bacteriophage showing the broadest host range covered 81% of all tested Streptococcus-isolates (30/37). Host ranges of Klebsiella pneumoniae- (9/26) and Pseudomonas aeruginosa-specific bacteriophages (14/33) covered a lower proportion of tested isolates. Seven bacteriophages (SEZ (n=3), KP (n=3), PA (n=1)) were selected according to theirhost ranges and submitted to electron microscopy for further characterization. Electron micrographs revealed that the selected bacteriophages belonged either to the family Demerecviridae (n=1), Myoviridae (n=1), Podoviridae (n=3) or Siphoviridae (n=2), which was confirmed by sequencing. Next, planktonic killing assay was conducted, incubating liquid cultures of Streptococcus equi ssp. zooepidemicus-, Klebsiella pneumoniae- and Pseudomonas aeruginosa-isolates in presence and absence of a specific bacteriophage and monitoring growth by optical density measurements for 24h. Significantly decreased bacterial growth, as determined by comparing the area under the curve (AUC), was observed in presence of 0/3 Streptococcus equi ssp. zooepidemicus-, 1/1 Pseudomonas aeruginosa- and 3/3 Klebsiella pneumoniae-specific phages, using a multiplicity of infection of 1 (MOI1; one phage per host cell). At MOI10 the most effective Streptococcus-phage reduced the AUC by 61%; the most effective Klebsiella-phage resulted in 95% reduction using MOI1 and for the Pseudomonas-phage AUC was reduced by 14% at MOI100. In conclusion, isolation of phages specific to equine uterine pathogens resulted in 28 purified phages. Seven phages were selected and further characterized by efficacy testing, sequencing and electron microscopy. Results of the in vitro host range determination and planktonic killing assay are promising, suggesting further investigations and the development of an alternative treatment approach for uterine infections in mares.

Antimicrobial and immunoregulatory effects of Lactobacillus delbrueckii 45E against genitourinary pathogens

BackgroundLactobacilli are essential microbiota that maintain a healthy, balanced vaginal environment. Vaginitis is a common infection in women during their reproductive years. Many factors are associated with vaginitis; one of them is the imbalance of microbiota in the vaginal environment. This study aimed to evaluate the antimicrobial properties of Lactobacillus delbrueckii 45E (Ld45E) against several species of bacteria, namely, Group B Streptococcus (GBS), Escherichia coli, Klebsiella spp., and Candida parapsilosis, as well as to determine the concentration of interleukin-17 (IL-17) in the presence of Ld45E.MethodsThe probiotic characteristics of Ld45E were evaluated by examining its morphology, pH tolerance, adhesive ability onto HeLa cells, hemolytic activity, antibiotic susceptibility, and autoaggregation ability. Then, the antimicrobial activity of Ld45E was determined using Ld45E culture, cell-free supernatant, and crude bacteriocin solution. Co-aggregation and competition ability assays against various pathogens were conducted. The immunoregulatory effects of Ld45E were analyzed by measuring the proinflammatory cytokine IL-17. A p-value less than 0.05 was considered statistical significance.ResultsLd45E is 3–5 mm in diameter and round with a flat-shaped colony. pH 4 and 4.5 were the most favorable range for Ld45E growth within 12 h of incubation. Ld45E showed a strong adhesion ability onto HeLa cells (86%) and negative hemolytic activities. Ld45E was also sensitive to ceftriaxone, cefuroxime, ciprofloxacin, and doxycycline. We found that it had a good autoaggregation ability of 80%. Regarding antagonistic properties, Ld45E culture showed strong antimicrobial activity against GBS, E. coli, and Klebsiella spp. but only a moderate effect on C. parapsilosis. Cell-free supernatant of Ld45E exerted the most potent inhibitory effects at 40 °C against all genital pathogens, whereas bacteriocin showed a robust inhibition at 37 °C and 40 °C. The highest co-aggregation affinity was observed with GBS (81%) and E. coli (40%). Competition ability against the adhesion of GBS (80%), E. coli (76%), Klebsiella (72%), and C. parapsilosis (58%) was found. Ld45E was able to reduce the induction of the proinflammatory protein IL-17.ConclusionsLd45E possessed antimicrobial and immunoregulatory properties, with better cell-on-cell activity than supernatant activity. Thus, Ld45E is a potential probiotic candidate for adjunct therapy to address vaginal infections.

Analyses of human papillomavirus, Chlamydia trachomatis, Ureaplasma urealyticum, Neisseria gonorrhoeae, and co-infections in a gynecology outpatient clinic in Haikou area, China

BackgroundThe purpose of this study was to study the infection rates of Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU), Neisseria gonorrhoeae (NG), and co-infections with human papillomavirus (HPV) in a hospital gynecology outpatient clinic in the Haikou region in 2021.MethodsFrom January to December 2021, the Women and Children Medical Center of Hainan Province collected 2389 samples of cervical exfoliated cells and vaginal swab specimens from gynecologic outpatients. The samples were then analyzed descriptively for data, and the detection rate of each pathogen was tallied. All vaginal swabs were obtained for CT, UU, and NG DNA testing, and cervical exfoliated cells for HPV genotyping. Analyses were performed on the detection rate of each group.ResultsIn 2389 samples, the frequencies of pathogen identification among the 2389 samples were as follows: UU (58.43%); HPV (17.29%); CT (7.99%); and NG (0.38%). HPV, CT, UU, and NG were detected in 33.33%, 22.55%, 77.45%, and 2.94% of individuals between 15 and 20 years of age, respectively. The detection rates of CT, UU, and NG were substantially greater in the HPV-positive group than the the HPV-negative group (P < 0.05).ConclusionAmong gynecologic outpatients at a hospital in the Haikou area, the probability of mixed infections with genital tract pathogens in HPV-positive patients was higher compared to HPV-negative patients. Reproductive tract infections are becoming more prevalent in younger people, hence adolescent sexual health education needs improvement.

Determinants of Acquisition, Persistence, and Clearance of Oncogenic Cervical Human Papillomavirus Infection in the Philippines Using a Multi-Omics Approach: DEFEAT HPV Study Protocol

HPV infection is one of the most studied risk factors in cervical cancer-the second most common cancer site and cause of death due to cancer in the Philippines. However, there is a lack of population-based epidemiological data on cervical HPV infection in the Philippines. Local reports on co-infections with other lower genital tract pathogens, commonly reported globally, are also lacking, which emphasizes the need to increase efforts in targeting HPV prevalence, genotype, and distribution. Hence, we aim to determine the molecular epidemiology and natural history of HPV infection among reproductive-age Filipino women using a community-based prospective cohort design. Women from rural and urban centers will be screened until the target sample size of 110 HPV-positive women (55 from rural sites and 55 from urban sites) is reached. Cervical and vaginal swabs will be collected from all screened participants. For HPV-positive patients, HPV genotypes will be determined. One hundred ten healthy controls will be selected from previously screened volunteers. The cases and controls will comprise the multi-omics subset of participants and will be followed up after 6 and 12 months for repeat HPV screening. Metagenomic and metabolomic analyses of the vaginal swabs will also be performed at baseline, after 6 months, and after 12 months. The results of this study will update the prevalence and genotypic distribution of cervical HPV infection among Filipino women, determine whether the current vaccines used for HPV vaccination programs capture the most prevalent high-risk HPV genotypes in the country, and identify vaginal community state types and bacterial taxa associated with the natural history of cervical HPV infection. The results of this study will be used as the basis for developing a biomarker that can help predict the risk of developing persistent cervical HPV infection in Filipino women.

Innate Lymphoid Cell Plasticity in Mucosal Infections.

Mucosal tissue homeostasis is a dynamic process that involves multiple mechanisms including regulation of innate lymphoid cells (ILCs). ILCs are mostly tissue-resident cells which are critical for tissue homeostasis and immune response against pathogens. ILCs can sense environmental changes and rapidly respond by producing effector cytokines to limit pathogen spread and initiate tissue recovery. However, dysregulation of ILCs can also lead to immunopathology. Accumulating evidence suggests that ILCs are dynamic population that can change their phenotype and functions under rapidly changing tissue microenvironment. However, the significance of ILC plasticity in response to pathogens remains poorly understood. Therefore, in this review, we discuss recent advances in understanding the mechanisms regulating ILC plasticity in response to intestinal, respiratory and genital tract pathogens. Key transcription factors and lineage-guiding cytokines regulate this plasticity. Additionally, we discuss the emerging data on the role of tissue microenvironment, gut microbiota, and hypoxia in ILC plasticity in response to mucosal pathogens. The identification of new pathways and molecular mechanisms that control functions and plasticity of ILCs could uncover more specific and effective therapeutic targets for infectious and autoimmune diseases where ILCs become dysregulated.

Virulence Characterization of Coagulase-Negative Staphylococci Isolated from Sexually Active Women with Symptomatic Genital Tract Infection, In Comparison with Staphylococcus aureus Isolated from the Same Cases

This study included 33 coagulase-negative staphylococci (CNS) and 35 Staphylococcus aureus isolated from sexually active females with symptomatic genital tract infection (64 pregnant and 110 non-pregnant). Comparison of phenotypic characteristics showed that the difference is not significant between CNS and S. aureus isolates for hemolytic activity (63.6% and 54.2%, respectively); lipolytic activity (87.8% and 88.5%, respectively); protease production (18.1% and 31.4%, respectively). and biofilm formation ( 96.9% and 94.2%, respectively). Whereas the difference is significant (P≤ 0.05) for urease production (18.1% and 42.8%, respectively). With the exception of lipolytic activity and biofilm formation, co-expression of other factors showed that there was an opposite expression of hemolytic and proteolytic activities of both CNS and S. aureus isolates. For CNS only 3/33 (9%) of the isolates were hemolytic and preoteolytic while for S. aureus only 2/35 (5.7%) isolates had this characteristics. Also for S. aureus isolates, ureolytic activity was more correlated with hemolytic (20%) and proteolytic (11.4%) activities than in CNS (6.0% and 3.0%, respectively). We concluded that, for this study included factors, the virulence of CNS isolated from females' genital tract infection is comparable to that of the potential females' genital tract pathogen, S. aureus. So that, CNS can be opportunistic pathogens in these cases rather than mere contaminants.

Molecular Epidemiology of Genital Infections in Campania Region: A Retrospective Study.

This study provides updated information on the prevalence and co-infections caused by genital microorganisms and pathogens: Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, Ureaplasma urealyticum, Trichomonas vaginalis, and Gardnerella vaginalis, by retrospectively analyzing a cohort of patients living in the Naples metropolitan area, Campania region, Southern Italy. To investigate the genital infections prevalence in clinical specimens (vaginal/endocervical swabs and urines) collected from infertile asymptomatic women and men from November 2018 to December 2020, we used a multiplex real-time PCR assay. Of the 717 specimens collected, 302 (42.1%) resulted positive for at least one of the targets named above. Statistically significant differences in genital prevalence of selected microorganisms were detected in both women (62.91%) and men (37.08%). G. vaginalis and U. parvum represented the most common findings with an 80.2% and 16.9% prevalence in vaginal/endocervical swabs and first-voided urines, respectively. Prevalence of multiple infections was 18.18% and 8.19% in women and men, respectively. The most frequent association detected was the co-infection of G. vaginalis and U. parvum with 60% prevalence. Our epidemiological analysis suggests different infection patterns between genders, highlighting the need to implement a preventative screening strategy of genital infections to reduce the complications on reproductive organs.

STATE OF CYTOKINE PROFILE IN PATIENTS WITH ADENOMYOSIS

Introduction. Adenomyosis is an important problem of modern gynecology, which causes significant fertility declines, leads to disability, dysfunction of adjacent organs, decreased quality of life of women. Objective: to study the state of cytokine profile in patients with adenomyosis.Material and methods. 35 female patients of whom 20 (57.1 %) with adenomyosis of I degree (study group) and 15 (42.9%) healthy female patients (control group) were examined. We determined the content of cathelicidin (LL-37) and β-defensin IL-1β, 4, 6, 10, 18, TFN-α, VEGF in blood serum, cervical mucus and uterine lavage by immunoenzyme method. To assess cytokine imbalance, we used a ratio between IL-1β/IL-10, IL-6/IL-10, TFN-α/IL-10, IL-4/ IFN-γ. Local immunity was studied in women depending on the presence of genital pathogens. In cervical mucus, we determined the presence of HSV-1.2, CMV by the PCR method. In peripheral blood - the presence of HSV-1.2, CMV specific IgM, IgG.Results. Determination of DNA of HSV1.2, CMV pathogens by PCR analysis of cervical mucus of patients with adenomyosis showed the presence of mono-viral contamination of HSV1.2 and CMV in 40 % of cases. Moreover, HSV1.2 was detected 1.7 times more frequently than CMV (p&lt;0.05). The proportion of mixt-viral contamination (HSV1.2+CMV) was 20 %.Six (30 %) patients with adenomyosis had an increased IgG titer against HSV1,2, five (25 %) had an increased IgG titer against CMV. No IgM against HSV1.2, CMV was detected. The study of cytokine profile in blood serum of patients with adenomyosis revealed a statistically significant increase in the level of pro-and anti-inflammatory cytokines. The increase in proinflammatory cytokines was less significant, and the concentration of anti-inflammatory cytokines was more significant, especially IL-10, indicating a cytokine imbalance with Th2-type cytokines predominating. In addition, in women with adenomyosis, an increase in the concentration of pro-and anti-inflammatory cytokines was observed in cervical mucus and uterine lavage, and the dynamics correlated with the values in blood serum; the degree of increase exceeded the values in blood serum. An increase of angiogenesis factor (VEGF) both in blood serum and locally may contribute to activation of vascular growth and ectopic endometrium. On the one hand, antimicrobial peptides decrease in blood serum and to a greater extent locally (defensin in cervical mucus: 384.5±39.1 pg/ml, in uterine lavage 252.2±24.8 pg/ml; in the control group – 447.1±45.3 pg/ml, 446.2±39.3 pg/ml, respectively; ±6.8 pg/ml, in the control group – 62.2±6.1 pg/ml. In all probability, these immune changes through the mechanism of epithelial-mesenchymal transition stimulate the migration of endometrial cells to the myometrium; endocrine imbalance may also contribute to this.Conclusions. The results of the studies showed a pronounced imbalance of the cytokine profile in patients with adenomyosis in case of genital infection. Immune disorders formed against the background of viral pathogens can contribute to the penetration of the endometrium into the myometrium and the development of the disease. The subject for further research should be the search for adequate therapeutic approaches aimed at the regulation of increased proliferation and invasion of the endometrium.

Emergence of genital infections due to Haemophilus pittmaniae and Haemophilus sputorum

BackgroundWe report for the first time the association of Haemophilus pittmaniae and Haemophilus sputorum with urethritis in men who have sex with men and who engage in unprotected intercourse. Patients and methodsA search for genital pathogens was conducted using urethral exudate cultures and PCR tests for Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma spp., and Ureaplasma spp. Recovered microorganisms were identified by MALDI-TOF mass spectrometry, and their susceptibility was evaluated by diffusion gradient test. ResultsH. pittmaniae and H. sputorum were isolated. They both proved susceptible to ampicillin, cefixime, and trimethoprim/sulfamethoxazole. H. pittmaniae was also susceptible to tetracycline and H. sputorum to moxifloxacin. ConclusionThe increased frequency of potentially resistant Haemophilus spp. isolates in genital exudates highlights the need for greater surveillance of these microorganisms and for their consideration in the differential diagnosis of genital system infections.

665 The state of local immune interrelations in patients with adenomyosis

ObjectiveThe frequency of adenomyosis reaches 70%. The intrusion of the basal layer of the endometrium into the myometrium is possible in case of a violation of local immunity, which is formed against the background of causative agents of genital infection. The study of immune interrelations in the uterus in patients with adenomyosis will expand the understanding of the pathogenesis. Study design: 159 women with adenomyosis were examined. In the cervical secretion, a quantitative assessment of the levels of cytokines associated with the activity of innate immune defense IL-4,8,10,18, TNF, INF, the concentration of sIgA, IgA, M, G, the concentration of C3, C4 components of complement and lysozyme. In the endometrium, Chlamydia trachomatis, Ureaplasma spp., Mycomlasma genitalium, HSV1,2 / CMV, HPV were determined by PCR. Results: In the presence of genital pathogens in patients with adenomyosis, disorders of innate immunity were revealed, manifested in an increased content of sIgA, IgA, G (p <0.05), which may be a consequence of impaired permeability of the mucous membranes. An increase in the level of pro-inflammatory (IL-8, IL-18, TNF) and a decrease in the level of anti-inflammatory cytokines (IL-4, IL-10), INF (p <0.05) were revealed. The complement system is activated, as evidenced by an increase in the level of C3, C4 (p <0.05). An increase in proinflammatory cytokines at the local level aims to limit inflammatory processes in the endometrium and promoting the development of a T-cell immune response. Conclusions: In patients with adenomyosis, signs of immunopathology are observed, leading to a weakening of immune control, which creates conditions for the development of endometrial heterotopies. Formed against the background of bacterial and viral pathogens, immune disorders can contribute to the invasion of endometrial cells into the adjacent myometrium.

The Vaginal Microbiota Composition and Genital Infections during and after Pregnancy among Women in Pemba Island, Tanzania.

We investigated the vaginal microbiota (VMB) composition, prevalence of genital pathogens and their association among pregnant and post-delivery women in Pemba Island, Tanzania. Vaginal swabs were collected from 90 women, at two time points during pregnancy (<20 weeks of gestational age [GA] and ≥20 weeks GA) and once after delivery, when possible. IS-pro assay was used for VMB characterization. Chlamydia trachomatis (CT), Neisseria gonorrhea (NG), Trichomonas vaginalis (TV), Mycoplasma genitalium (MG) and human papillomavirus (HPV) were detected by qPCRs. VMB were mostly Lactobacillus dominant during pregnancy and non-Lactobacillus dominant post-delivery. A significant decrease in VMB richness was observed during pregnancy among paired and unpaired samples. Shannon diversity was significantly lower during pregnancy than post-delivery among unpaired samples. Klebsiella species and Streptococcus anginosus were the most commonly identified pathobionts at all timepoints. A high abundance of pathobionts was mostly seen in women with non-Lactobacillus dominant VMB. At ≥20 weeks GA timepoint during pregnancy, 63.0% of the women carrying one or more genital pathogen (either HPV, CT, TV, or MG) had L. iners dominant VMB. NG was not detected pre-delivery. This study contributes evidence on VMB composition, its changes during pregnancy and post-delivery, and their association with pathobionts and genital pathogens.