Vaginal Swab Samples Research Articles

Clinical Validation and Comparison of Bacterial Vaginosis Detection

Abstract Introduction/Objective Bacterial vaginosis (BV) is considered a top reason for women’s gynecological visits, causing symptoms such as vaginal discharge, foul odor, itching, and burning associated with the alteration of the normal vaginal microbiome. It is often presented as a decrease in Lactobacillus spp. and an increase in anaerobic organisms such as Gardnerella vaginalis and Atopobium vaginae. In response to the demand for more automated and less subjective testing, molecular methods have recently been developed to aid in BV testing. The objectives of this study are to validate and compare the molecular Aptima® Bacterial Vaginosis Assay by Hologic® to the established BV Screen via a Gram stained vaginal smear graded through Nugent scoring at RUSH University Medical Center in Chicago, IL and perform a cost analysis between the testing methods. Methods/Case Report Samples of known BV value were tested on the Panther® instrument to validate the Aptima® Bacterial Vaginosis Assay for accuracy and precision. Clinical samples of vaginal swabs were used to make a Gram stained BV screen for Nugent scoring, then afterwards were placed into Hologic Multitest Aptima® probes for molecular testing on the Panther® instrument. Nugent scoring was performed blind by the molecular test. Molecular results of negative, positive, and invalid were then compared to the Nugent scores: scores of 0-3 for negative/normal vaginal flora, 7-10 for positive BV, and 4-6 for intermediate/altered scores were screened for discrepancy. Results (if a Case Study enter NA) The Aptima® BV Assay tested 52 known samples with 97% accuracy and 100% precision. A total of 80 clinical samples of vaginal swabs used for Gram stained BV screens were run on the Aptima® BV Assay; 61 provided conclusive data for comparative analysis. When compared to the original Nugent scores, the positive percent agreement (PPA) was 77.0% and negative percent agreement (NPA) was 92.9%. Conclusion The Aptima® BV Assay proved to be both accurate and precise in its validation testing. Clinically, it was able to screen for BV in vaginal samples with a good percent agreement to Gram stained BV screens. Further testing of additional clinical samples are to be done to continue analysis. Although the supplies needed to perform a Gram stain cost less than molecular testing, the reimbursement payment for molecular testing is much greater and overall reduces employee time costs for performing the test, making it the more cost effective choice for the laboratory.

Frequent Evaluation of Herpes Simplex Virus Type 2 in Women with Genital Herpes by Realtime PCR

Background: Herpes simplex type 2 is a common infection worldwide. This disease is common in both developed and developing countries. Early detection of infection is very important to reduce the risk of infection. Real-time reliable PCR is a very sensitive and specific method that can be used as the best marker in determining the therapeutic effect by identifying a viral genome in an individual. The prevalence of herpes simplex virus type 2 in women with genital herpes was evaluated by Real time PCR method. Methods: From January 1999 to March 2010, 45 samples of vaginal swabs and cervix of women with genital herpes were examined for HSV virus DNA detection using Real Time PCR. Results: The mean age of the patients was 35.9 + 5.9. The percentage of positive cases of herpes simplex virus type 2 in the studied women was 22.2% and the history of infection with hpv was 33.3% vs. 12.5%. = 0.094 which was significant. Conclusion: Clinical specimens of vaginal swabs from genital herpes caused by herpes simplex virus 2 can be quantitatively analyzed instead of nucleic acid extraction and amplification by PCR.

Evaluation of the Causative Microorganisms and Antibiogram Results in Women with Vaginal Swab Culture Taken with Prediagnosis of Vaginitis

Aim: Vaginitis occurs when the vaginal flora is disturbed. This study aimed to evaluate the demographic data, causative microorganisms, and antibiogram results of women who underwent vaginal swab culture with a prediagnosis of vaginitis. Methods: Vaginal swab samples from 314 women who visited the Kafkas University Department of Obstetrics and Gynecology were included in the study. Women with immunosuppressive diseases, gynecological malignancies, and IUD use were excluded. Vaginal discharge or swab samples were Gram stained and cultured on various media. Microorganisms were identified using conventional and biochemical methods, and antibiotic susceptibility tests were performed using the Kirby-Bauer disk diffusion method, according to the EUCAST guidelines. Statistical analyses were performed using SPSS, with categorical variables assessed using the chi-square test or Fisher's exact test, and continuous variables assessed using the Mann-Whitney U test. The study complied with the recommendations of the Declaration of Helsinki for human biomedical research and was approved by the Non-Interventional Clinical Research Ethics Committee of the Kafkas University Faculty of Medicine. Results: The median age of 314 women included in this study was 39 (20-69) years. . In total, 73.6% (n=231) were premenopausal, 24.5% (n=77) were postmenopausal, and 1.9% (n=6) were pregnant. The most common complaint was abdominal/pelvic pain (142, 45.2%). Vaginal cultures from 123 women (39.1%) showed the growth of different microorganisms. The most common causative microorganism was Escherichia coli (41.5%). According to the antibiogram results, the most sensitive antibiotic was gentamicin (65.9%), and ampicillin (52.8 %) was the most resistant. Conclusion: It is important to quickly and accurately identify microorganisms involved in the etiology of the correct treatment.

Evaluation of the Prevalence, Antimicrobial Resistance Trait, and Virulence Determinants in Staphylococcus aureus Isolates from the Anogenital Area of 35-37 Weeks Pregnant Women

Staphylococcus aureus is a major pathogen in the postpartum period being frequently implicated as a cause of mastitis, and nursery outbreaks, among others as well as its increasing resistance to common antibiotics. This study therefore aimed at evaluating the presence of S. aureus in pregnant women attend¬ing clinics in rural towns in Eastern Cape, South Africa. A total of 82 pregnant women at 35‒37 weeks of gestation were sampled for the presence of S. aureus. Using standard microbiological methods, presump¬tive S. aureus isolates were obtained from vaginal and rectal swab samples. The isolates were confirmed by conventional polymerase chain reaction and then tested for antimicrobial susceptibility using the disc diffusion technique. Presence of resistance genes and virulence determinants were equally profiled. A to¬tal of 47 (57.3%) out of 82 pregnant women were colonized with S. aureus. Only two virulence genes PVL- 37/47 (78.7%) and eta-10/47 (21.3%) were detected in the isolates. The isolates were all resistant to penicillin G and clindamycin (100%), while resistances to tetracycline, vancomycin, rifampicin were 71% and resistance to oxacillin and erythromycin were above 80% while resistance to other antibiotics tested were below 40%. The isolates showed multiple resistance to 5-6 antimicrobials with indices ranging from 0.5-0.6. Genes encoding resistance to erythromycin (ermB), tetracycline (tetM), and rifampicin (rPOB) were found in 72.7% (34/47) of the isolates, while 15% (7/47) possessed the Bla-Z (penicillin). The high antibiotics resistance traits found in the isolates analysed indicates limited therapeutic options and portents a major threat to public health.

PSV-16 Evaluation of upper respiratory and fecal microbiomes in neonatal beef calves

Abstract There is a critical lack of information on the timing of colonization of bovine respiratory disease-related bacteria in the upper respiratory tract, potentially affecting calf health and disease onset. The objective of this study was to evaluate the upper respiratory tract and fecal microbiomes of neonatal beef calves during the first 24 h of life compared with their dams at the time of birth. Late-gestation commercial beef cows (n = 28) were assigned to individual, soil-surfaced pens (6.25 x 2.29 m) fitted with canvas tarps on shared fences to limit contact between unrelated pairs. Left nasal (LN), right nasal (RN), fecal, and vaginal swab samples were collected from cows at a single time point promptly after parturition. Swabs of the LN, RN, and rectum were collected from calves at birth (0 h), 6, 12, and 24 h post-parturition. Samples were flash-frozen and stored at -80°C until processing. DNA was extracted from swabs using Power Soil Pro kit. Extracted DNA was used to prepare DNA libraries for full-length 16S rRNA gene sequencing on a MinION Mk1C device. Sequenced reads were classified using Centrifuge and used in microbiome analyses conducted in R to estimate the changes in the microbiome according to animal type (cow versus calf), time of collection, and sample type (nasal, fecal, or vaginal). Upper respiratory microbiomes significantly differed (P < 0.005) between cows and their offspring at birth. In nasal microbiomes, animal type explained 12.6% (P = 0.002) and 18.5% (P = 0.001) of variance in beta diversity in left and right cavities, respectively. Animal type explained 42.1% of the beta diversity variance in fecal microbiomes (P = 0.001). Nasal microbiomes at birth were significantly more dispersed in dams compared with their calves in both the LN (P = 0.03) and RN (P = 0.03). Similarly, fecal microbiomes of dams were significantly more dispersed (P < 0.0001) than those of their offspring. There was a clear transition of the microbial communities associated with calf age in the upper respiratory and fecal niches. Calf age accounted for 2% (P = 0.02), 3.5% (P = 0.001), and 28% (P = 0.001) of the variance in the beta diversity of the LN, RN, and fecal microbiomes, respectively. After adjusting for calf age, the laterality of nasal microbiomes was not significantly (P = 0.682) associated with the composition of microbial communities. In conclusion, calf microbial communities were dissimilar to that of their dam at birth. Additionally, microbial community transition was evident with each subsequent sampling time point, potentially influenced by the presence of the dam.

Toward Analysis at the Point of Need: A Digital Microfluidic Approach to Processing Multi-Source Sexual Assault Samples.

Forensic case samples collected in sexual assaults typically contain DNA from multiple sources, which complicates short-tandem repeat (STR) profiling. These samples are typically sent to a laboratory to separate the DNA from sperm and non-sperm sources prior to analysis. Here, the automation and miniaturization of these steps using digital microfluidics (DMF) is reported, which may eventually enable processing sexual assault samples outside of the laboratory, at the point of need. When applied to vaginal swab samples collected up to 12h post-coitus (PC), the new method identifies single-source (male) STR profiles. When applied to samples collected 24-72h PC, the method identifies mixed STR profiles, suggesting room for improvement and/or potential for data deconvolution. In sum, an automated, miniaturized sample pre-processing method for separating the DNA contained in sexual assault samples is demonstrated. This type of automated processing using DMF, especially when combined with Rapid DNA Analysis, has the potential to be used for processing of sexual assault samples in hospitals, police offices, and other locations outside of the laboratory.

Molecular Characterization of Human Papilloma Virus among Women Attending Selected Hospitals in Sokoto, Sokoto State, Nigeria

Human Papilloma Virus (HPV) infection is the primary cause of virtually all cervical cancers. Cervical cancer despite being largely preventable is still the leading cause of gynaecological cancer related death among females in developing countries. This study was carried out to molecularly characterized Human Papilloma Virus among women attending selected hospitals in Sokoto, Sokoto State, Nigeria. This study involves the analysis of high vaginal swab obtained from 208 women. The high vaginal swab samples were analysed using PCR, DNA sequencing and phylogenetic analysis of the identified genotypes. Only two Human papilloma virus genotypes (HPV type 18 and 58) were detected in this study with Human papilloma virus type 58 as the most prevalent. This study identified only two Human papilloma virus genotypes (HPV types 18 and 58) in Sokoto, Nigeria which can serve as basis for establishing vaccine in the study area

Bacterial vaginosis (BV) and Trichomonas vaginalis (TV) co-infection, and bacterial antibiogram profile of pregnant women studied in Lagos, Nigeria

AimThis study was undertaken to determine the prevalence of Bacterial Vaginosis (BV), Trichomonas Vaginalis (TV) co-infection, and the antibacterial sensitivity profile of bacterial isolates.MethodsThe study was a cross-sectional study of 232 pregnant women on a routine antenatal visit between April 2019 and Sept. 2020, at Amukoko clinic in Lagos, Nigeria. The gynaecologist conducted the clinical examination on each patient looking for vaginal discharge and its consistency/homogeneity, colour and odour. Two High Vaginal Swab (HVS) samples were taken from every patient and a semi-structured questionnaire was used to gather the socio-demographic, practices/attitudes, and clinical information of each participant. One sample was employed for wet preparation to identify the TV and BV diagnosis using Amsel’s criteria and Whiff’s test. The second sample was used for bacterial culture and antibiogram was conducted using the disc diffusion technique. The Clinical Laboratory Standard Institutes’ (CLSI) interpretative criteria were used to categorise the results.ResultsThe mean age of the clients was 28.11 ± 7.08 years of age. The majority (88%) were aged 15–35 years. Only 81 (34.9%) had microbial organisms isolated or seen from their specimens and 19 (8.2%) of such were classified as having BV (Bacteriods or Gardnerella isolated). Of the 81 infected, 33 (40.8%) had only bacterial infection, 36 (44.4%) had TV alone and 12 (14.8%) had bacteria co-infected with TV. From the clinical records, the population that was classified as having UTI or vaginitis was only 46 (20.7%) The study observed age (15–35 years) related association between vaginosis/ TV co-infection (X2 = 7.9; P = 0.005). Participants with symptoms of vaginitis or UTI (mainly E. coli & pseudomonas spp. isolated), BV/co-infection with TV significantly associated with female traders (X2 = 8.5; P = 0.003) and were more associated with those from polygamous relationships (X2 = 18.79, P = 0.0001). Women in their 3rd and 2nd. trimester were more significantly associated with vaginal infection (X2 = 9.47, P = 0.002; X2 = 4.79, P = 0.029) respectively. The Pseudomonas showed susceptibility to ciprofloxacin (CIP) and cefuroxime (CXM). While, E. coli isolates were susceptible to cefepime, ciprofloxacin, and imipenem.ConclusionThere is a relatively low prevalence of BV and flagellate co-infection in the community studied.RecommendationWe recommend screening of antenatal women with underlying symptoms for BV and flagellates co-infection to avoid its progression to vaginitis.

Identification and Antimicrobial Susceptibility Patterns of Neisseria gonorrhoeae, Ureaplasma spp. and Mycoplasma spp. Isolated from Tribal Women

Sexually transmitted infections (STIs) are a major public health problem worldwide with significant social and economic implications. Effective control and prevention strategies necessitate a thorough understanding of the prevalence, isolation, and identification of STI pathogens. The present study aims to provide a comprehensive analysis of the isolation, identification, prevalence, and antimicrobial susceptibility pattern of STI pathogens based on culture method analysis. Endocervical /vaginal swab samples from female patients symptomatic for STI were cultured on different selective and differential media and pathogens were identified by colony morphology and biochemical tests. Antimicrobial Susceptibility Test (AST) of isolated and identified culture pathogen was performed by using Kirby-Bauer disc diffusion method. Among 209 endocervical/vaginal swab samples from symptomatic patients, 126 (60.28%) tested positive and 83 (39.71%) negative. Ureaplasma spp. (n = 100) was the most prevalent isolate, constituting 79.36% of culture-positive samples, followed by N. gonorrhoea (n = 99) at 78.57%, and Mycoplasma spp. (n = 41) at 32.54% individually and in combination. AST analysis revealed erythromycin (74%), ofloxacin (69%), and roxithromycin (64%) as the most resistant antibiotics for Ureaplasma spp. N. gonorrhoea showed the highest resistance to cefixime (78.79%), followed by ofloxacin (75.76%) and erythromycin (69.7%). Azithromycin and erythromycin exhibited 100% resistance against Mycoplasma spp. The study provides information on the prevalent bacterial pathogens involved in STIs among women in Anuppur and Shahdol districts, Madhya Pradesh. Understanding the diversity, distribution patterns and antibiotic sensitivity of these pathogens is crucial for developing targeted interventions and effective prevention strategies in such resource-limited areas.

Resistant C. albicans implicated in recurrent vulvovaginal candidiasis (RVVC) among women in a tertiary healthcare facility in Kumasi, Ghana

BackgroundVulvovaginal candidiasis is a common fungal infection that affects the female lower genital tract. This study determined the major risk factors associated with vulvovaginal infection (VVI) in the Ashanti region of Ghana and also determined the antifungal resistance patterns of Candida albicans isolates to some antifungals.MethodsThree hundred and fifty (350) high vaginal swab (HVS) samples were collected from women who presented with signs and symptoms of VVI. A structured questionnaire was administered to one hundred and seventy-two (172) of the women. HVS samples were cultured on Sabouraud dextrose agar with 2% chloramphenicol. The polymerase chain reaction was employed to confirm C. albicans. Antifungal susceptibility testing was performed and the susceptibility of C. albicans isolates to fluconazole, clotrimazole, amphotericin B, nystatin, miconazole and 5-flurocytosine were assessed.ResultsVaginal infection was most prevalent amongst females in their reproductive age (21 to 30 years; 63.0%). The study found a significant association between vaginal infections and some risk factors such as sexual practices (p < 0.001), antibiotic misuse (p < 0.05), poor personal hygiene (p < 0.005) and birth control methods (p < 0.049). Out of the 350 HVS samples collected, 112 yielded yeast cells with 65 (58%) identified as C. albicans. The C. albicans isolates were resistant to 5’ flucytosine (100%), fluconazole (70%), voriconazole (69.2%), miconazole (58.5%) and nystatin (49.2%). C. albicans isolates were more susceptible to amphotericin B (53.8%) and clotrimazole (45.1%), although an appreciable number of isolates showed resistance (46.1% and 52.3%, respectively).ConclusionThere should be nationwide education on all associated risk factors of VVI. Also, use of the various antifungal agents in vaginal candidiasis should proceed after antifungal susceptibility testing to ensure efficacious use of these agents.

Molecular Detection of Coxiella burnetii in Vaginal Swab Samples from Sheep That Aborted.

Background: Coxiella burnetii, an obligate intracellular bacterium, is the etiological agent of Q fever in humans and one of the causes of abortion in small ruminants. Although coxiellosis is considered an exotic disease, there are a few reports in Mexico. Methods: The objective of this work was to determine the presence of C. burnetii DNA in vaginal samples from sheep that presented abortion and ram semen. A total of 180 vaginal exudate samples and 20 semen samples were obtained from five Central and Southern States of Mexico. Total DNA was extracted from vaginal swabs and C. burnetii was identified by PCR amplification and sequencing of the IS1111 insertion sequence. Results and Conclusion: In total, 110 (110/180) vaginal samples and 12 (12/20) semen samples were positive for C. burnetii. This is the first report of C. burnetii in sheep that aborted and in ram semen in Mexico.

Prevalence Studies on Brucellosis in Buffalo Population in Nearby Region of Hyderabad, Telangana

Background: Brucellosis in India is highly endemic in many states and reported as the most prevalent disease in dairy cattle. Brucellosis is a highly contagious bacterial disease caused by various Brucella species mainly affecting wide range of different species of animals like cattle, sheep, goat and pigs. The present study was undertaken to ascertain the prevalence of brucellosis in suspected buffalo population showing reproductive disorders in selected dairy farms residing nearby region of Hyderabad, Telangana. Methods: Total number of 204 vaginal swab samples were collected aseptically from suspected buffaloes in nearby private dairy farms and were screened by employing various diagnostic methods. Enzyme Linked Immuno Sorbent Assay (ELISA), Lateral Flow Assay (LFA) and molecular identification through Polymerase Chain Reaction (PCR) were conducted. The sensitivity of tests was recorded considering PCR as gold standard method. Result: Screening of 204 vaginal swab samples and prevalence of brucellosis were recorded by ensuring the positive reactions and also depicting the sensitivity of various diagnostic tests namely ELISA, LFA and PCR accounting 92(45.1%), 87 (42.64%) and 113(55.4%) samples respectively. The specificity of both LFA and ELISA were recorded 100% when compared to PCR assay.

O-276 Implantation failure is not associated with seven different vaginal bacterial community-state types; however, a distinct signature, including Ureaplasma parvum, is strongly linked to treatment failure

Abstract Study question Is microbial diversity or vaginal bacterial composition associated with embryonic implantation in women undergoing frozen-thawed embryo transfer? Summary answer While global diversity measures of microbial composition do not correlate with positive implantation, specific taxa may serve as predictors of treatment outcomes. What is known already Clinical studies have indicated a reduced likelihood of embryo implantation in women diagnosed with microbial dysbiosis. The application of Next-Generation Sequencing for microbiota analysis has significantly advanced our understanding of vaginal microbiome composition. Recent studies have proposed distinct vaginal microbiome patterns as potential predictors of IVF treatment success or failure. However, the evidence regarding the biological basis of test algorithms (such as the independent association of distinct community state types with outcomes) and the reliability of diagnostic test performance (including replicability and external validity) in predicting implantation success or failure in women undergoing embryo transfer remains less robust. Study design, size, duration In a prospective, single-centre observational cohort study (05/2017-03/2019, NCT03507673, local ethical approval reference no.18-005), conducted at a university-affiliated centre for reproductive medicine in Germany, vaginal swab samples were collected from infertile, asymptomatic female patients (n = 260), aged 18-45, undergoing frozen-thawed embryo transfer. Patients, each contributing a single treatment cycle, received sequential oral estradiol and oral estradiol/dydrogesterone in an HRT regimen for FET. Vaginal swabs were taken from baseline visit before transfer day and stored at –80 °C. Participants/materials, setting, methods 16S rRNA gene amplicon sequencing was performed targeting V3/V4 hypervariable region using Illumina MiSeq platform. Raw data assembly, filtering and taxonomic assignment were performed using mothur against EzBioCloud database. Statistical analyses, and visualization were conducted using R. Community structure via hierarchical clustering, alpha and beta diversity, and indicator species via LefSe analysis were assessed. Main results and the role of chance Based on a hierarchical clustering using the 25 most abundant bacterial taxa, seven separate community state types (CSTs) being either dominated by species of the genus Lactobacillus (CST-A, CST-B, CST-C, CST-E, CST-G) or Gardnerella (CST-F) or presenting a variable community composition (CST-D) were identified. A community state type dominated by Lactobacillus fornicalis was found in a small fraction of women and enhanced resolution among the Gardnerella genus impacts CST definitions in this sample. Embryonic implantation had no significant association to either alpha diversity measures or CST distribution; likewise, the previously suggested ratio of relative abundances of Lactobacillus iners vs. Lactobacillus crispatus was not associated with implantation. Lactobacillus fornicalis is positively linked to implantation, while Lactobacillus iners and Ureaplasma parvum were linked to implantation failure: above 50% relative abundance of Lactobacillus fornicalis, was associated with 75% positive implantation. Conversely, just 0.5% relative abundance of Ureaplasma parvum was associated with a decrease in implantation rate to approximately 20%. LefSe and binary regression analysis identified a relatively rare (6% in our sample) microbiome signature, mostly from CST-C (Lactobacillus iners dominated) with a very low likelihood of implantation, with Ureaplasma parvum as the common denominator. Limitations, reasons for caution Studies on the vaginal microbiome in IVF face issues such as study design variability, small sample-sizes, limited control for temporal dynamics, and patient heterogeneity. Attempts at establishing causal relationships through interventional studies in IVF have been limited and inconclusive. Understanding the microbiome’s functional aspects is crucial for reproductive outcome elucidation. Wider implications of the findings Current study findings don’t confirm previously reported associations between common vaginal bacterial CSTs and outcome. Therefore, screening for vaginal microbiome composition in asymptomatic women with subfertility is not recommended outside research. Signatures/taxae with an independent and strong negative association with implantation are likely to be rare yet deserve further attention. Trial registration number NCT03507673

O-281 Characterization of Vaginal microbiota and reproductive outcomes associated with assisted reproductive technologies through next-generation sequencing, from Indian population

Abstract Study question Is there any association between the composition of the vaginal microbiota and the reproductive outcomes in infertile women’s undergoing for assisted reproductive treatments (ART)? Summary answer Higher microbial abundance in non-pregnant patients compared to pregnant patients. These pathogens could be used as a possible marker for a higher IVF failure rate. What is known already Various next- generation sequencing methods have been used to study bacterial populations in infertile Women. Vaginal microbiota could have an influence on success rates of fertility treatments. The bacterial microbiota varies by the population. There is no direct evidence on this subject to guide clinical decision making or its influence on the hierarchy of fertility treatments. While the presence of pathogenic bacteria non-Lactobacilli dominated microbiota is associated with poor reproductive outcomes in assisted reproductive treatments (ART). Additional studies, particularly those using metagenomics approaches, may be useful. Study design, size, duration A total of 240 infertile women’s were included in the study. The study population consists of patients attending to the ART unit of a tertiary medical center Udaipur, India, between November 2021 to October 2023. Depending on the biochemical pregnancy results, they were divided into two groups. Group 1 (n = 128) was pregnancy (P), and Group 2 (n = 112) non- pregnancy (NP) group, Patients were aged between (20-45 years). The ethical approval was obtained from the institute. Participants/materials, setting, methods DNA was extracted from vaginal swab samples collected at the time of oocyte pick up using the commercially available ( DNeasy Power Soil kit). The Vaginal microbiota of samples were analyzed using 16S ribosomal RNA gene amplification (MinION) Oxford Nanopore Ltd. Bioinformatics analysis was performed using QIIME2 and Microbiome Analyst packages. Alpha, beta diversity and taxonomic characterization were compared for microbial composition. The findings of the principle coordinate analysis were shown using R version 4.0. Main results and the role of chance Our results show that higher Shannon and Simpson index were obtained in the non- pregnancy (NP) group compared to the pregnancy (P) group. In addition, the partial least squares discriminant analysis (PLS-DA) revealed a difference in microbial composition between pregnancies (P) and non- pregnancy (NP) groups, as well as a higher microbial abundance in non-pregnant patients compared to pregnant patients. At the phylum level Firmicutes, Actinobacteria, Proteobacteria, Fusobacteria, and Bacteroidetes are the most common predominant in the pregnancy (P) group (80.77, 9.52, 9.18, 0.04, and 0.40%, respectively) and the non- pregnancy (NP) group (72, 61, 12.41, 8.01, and 3.75%). A higher abundance of Firmicutes (80.77 vs. 72.61%) and Proteobacteria (9.18 vs. 8.01%), and a lower abundance of Actinobacteria (9.52 vs. 12.41%, p = 0.03), Fusobacteria (0.04 vs. 8.01%, p = 0.02), and Bacteroidetes (0.40 vs. 3.75%, p = 0.03) were observed in pregnancy (P) group. At the genus level, pregnancy group (P) increased the abundance of probiotics lactic acid bacteria (74.61 vs 63.09%) and decreased the abundance of the pathogen Gardnerella (6.03 vs 7.24%, p = 0.03), Atopobium (0.84 vs 4.14%, p = 0.02), Sneathia (0.03 vs. 3.75%, p = 0.02), and Prevotella (0.38 vs. 3, 02%, p = 0.04). Limitations, reasons for caution The limited sample size is the primary constraint of this study. Larger studies with a larger sample size are needed to confirm the distinct microbiome patterns identified in female reproductive tracts samples in connection to infertility and live birth rate (LBR) outcomes. Wider implications of the findings Vaginal microbiota composition associated to different clinical outcomes in patients undergoing ART might have profound implications. This may lead to failed fertility treatments and reduced live birth rate (LBR). More studies are needed to clarify the relative effectiveness of treatments for infertility. Trial registration number NA

Benefits of maternal pectin supplementation in gestation diet on vaginal microbiota of sows and intestinal health of newborn piglets.

Pectin is a proven prebiotic and widely used in human health products. This study aims to assess the impact of dietary pectin supplementation during gestation on sow vaginal microbiota and the offspring's intestinal composition. Thirty sows were randomly allocated to two groups and fed a standard diet (CON) or a standard diet supplemented with 3 g/kg pectin (PEC). Blood, feces, and vaginal swab samples from the sows and blood, intestines issue, and colonic content samples from the offspring were collected and analyzed. The results indicate that the relative abundance of vaginal Lactobacillus was notably enhanced in the PEC group and fecal β-glucuronidase (β-G) activity and plasma 17β-estradiol (E2) concentration were also significantly increased in the PEC group. Newborn piglets were found to host different microbial communities as well. At the phylum level, Proteobacteria dominated in the CON group, and Firmicutes was predominant in the PEC group. Newborn piglets in the PEC group had a lower interleukin-6 (IL-6) concentration in their plasma. The expression of intestinal cytokines of offspring was improved as well. Villus height and villus height/crypt depth (V/C) in the PEC group were extremely higher than those in the CON group. In conclusion, dietary pectin supplementation can be of benefit to both sows and newborn piglets.

Combined activity of blue vitriol, brimstone and black stone on clinical Candida species isolates using fractional inhibitory concentration Index

Combination antimicrobial therapy (CAT) involves intentional addition of two or more antimicrobial agents for better therapeutic outcome. CAT is normally applied for treatment of polymicrobial infections, life-threatening infections and prevention of the emergence of resistance. This research aimed at determining the combined activity of blue vitriol, brimstone and black stone on clinical Candida species isolates using Fractional Inhibitory Concentration Index. The brimstone, blue vitriol and black stone samples were purchased from a community market in southeastern Nigeria. The isolates were obtained from high vaginal swab samples of patients attending a Teaching hospital in Nigeria and identified based on their morphological, physiological and molecular characteristics. The minimal inhibitory concentrations (MIC) of the samples were determined using the broth dilution method. The fractional inhibitory concentration indexes (FICI) of the samples against the isolates were obtained from their individual FIC values. FICI of &lt;1 indicates synergism; 1 to 2 (Indifference); while &gt;2 (antagonism). All the test agents were synergistic against C. albicans (FICI &lt; 1). All the test agents were indifferent against C. tropicalis (FICI 1 - 2). Brimstone + black stone combination was indifferent (FICI 1 - 2) against C. glabrata; while all the test agents were antagonistic against the isolate (FICI &gt; 2). Blue vitriol + Black stone combination was antagonistic (FICI &gt; 2) against C. parapsilosis; Blue vitriol + brimstone was indifferent (FICI 1 - 2); while brimstone + black stone was synergistic (FICI &lt; 1). The findings have revealed that antimicrobial combination outcome could be synergistic, antagonistic or indifferent; thus the need for proper in vitro screening before combination therapy.

Feasibility and acceptability of sexually transmitted infection screening during antenatal care of women in Dhaka, Bangladesh.

Sexually transmitted infections (STIs) are a major public health concern worldwide. Untreated STIs may have serious sequelae, particularly in pregnant women. The objective of this study was to assess the feasibility and acceptability of screening and treating common STIs in women during pregnancy in Bangladesh. Women were enrolled from four maternity clinics/hospitals serving the lower-middle class population in Dhaka, Bangladesh. The participants were interviewed, and vaginal swab samples were collected by clinical staff. Specimens were tested for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and high-risk Human Papilloma Viruses (HPVs) using GeneXpert (Cepheid, Sunnyvale, California). Women were informed of their test results and were provided treatment for curable infections. A test of cure was performed. Out of 1157 pregnant women approached, 1000 (86.4%) participated. Ninety-one percent women learned of their test results on the same day of testing. Out of the 996 valid results, 7 (0.7%) tested positive for Chlamydia trachomatis and 1 (0.1%) for Trichomonas vaginalis. There were no gonorrhoea cases. Out of the 971 women with valid results for high-risk HPVs, 46 (4.7%) tested positive. Screening women for STIs during antenatal care was highly feasible and well-accepted in Bangladesh. While the prevalence of common curable STIs was very low, hrHPV infection prevalence was moderately high. Our findings support period monitoring of STIs and continued prevention efforts for cervical cancer in Bangladesh.

Comparison of cervicovaginal fluid extracellular vesicles isolated from paired cervical brushes and vaginal swabs

AbstractEndometriosis is a common gynaecological condition, with a long diagnostic delay. Surgery is required to confirm a diagnosis, highlighting the need for a non‐invasive biomarker. Extracellular vesicles (EVs) may have a role in endometriosis pathogenesis, yet there is limited EV biomarker literature available. This study aimed to investigate the feasibility of isolating cervico‐vaginal fluid EVs sampled using cervical brushes and vaginal swabs and to compare these methods. After providing informed consent, patients undergoing surgery for suspected endometriosis had cervical brush and vaginal swab samples collected under general anaesthetic. Isolated EVs were characterised through negative stain transmission electron microscopy (TEM), Western blotting (TSG101, CD63, Calnexin, ApoB, Albumin), tunable resistive pulse sensing (TRPS), microBCA assays and RT‐qPCR of miRNAs. PCR was performed on samples prior to EV isolation to assess bacteria present in samples. Cervical brush and vaginal swab EVs were intact vesicles with limited co‐isolated contaminants. Cervical brushes had higher concentrations of particles compared to match vaginal swabs, although both samples had low concentrations. Protein and miRNA yield were similar between matched samples. PCR demonstrated only a small amount DNA within samples was bacterial (&gt;0.5%). Cervico‐vaginal fluids EVs were successfully isolated from cervical brushes and vaginal swabs, demonstrating a new method of sampling reproductive EVs. EV yield from both sample types was low. Similar protein and miRNA levels suggest either sampling method may be suitable for biomarker studies.

Occurrence of Coxiellosis in ruminants and its associated risk factors

Coxiellosis in animals is caused by the zoonotic pathogen, Coxiella burnetii. Although the disease is of public health importance it remains underdiagnosed and underreported. The cross- sectional study was aimed to estimate the occurrence of the disease in livestock of study area and also to identify the risk factors associated with the disease in animals. Blood, serum, and vaginal swabs samples were collected from 200 ruminants (cattle, sheep, and goats), across various farms in Karnataka, India. These samples were then screened using ELISA and PCR (com1 and IS1111). A questionnaire was administered to the farm owners to collect the risk factor-related information. About 5.26 % cattle, 12.3 % sheep, and 12.5 % goats were positive by ELISA. By PCR, 9.47 % cattle, 9.3 % sheep, and 10 % goats were positive. Overall, the occurrence of 14.73 %, 18.46 % and 17.5 % was estimated in cattle, sheep and goat, respectively. PCR targeting the IS1111 gene detected higher number of samples as positive as compared to the com1 gene PCR. Higher number of vaginal swab samples were detected as positive as compared to blood. History of reproductive disorders (OR: 4.30; 95 %CI:1.95- 9.46), abortion (OR: 30.94; 95 %CI:6.30- 151.84) and repeat breeding (OR:11.36; 95 %CI:4.16- 30.99) were significantly associated with coxiellosis (p < 0.005). Multivariable analysis by logistic regression model analysis suggested retained abortion, repeat breeding and rearing of animal in semi-intensive system as factors significantly associated with the infection. Cultural identification of the PCR positive samples were cultured using embryonated egg propagation and cell culture techniques and positivity was confirmed in six samples. Phylogenetic analysis of the com1 and IS1111 gene revealed clustering based on similar geographic locations. The study estimated the occurrence of the disease in the study area and identified the potential risk factors.

Socio-demographic, obstetric characteristics and molecular confirmation of &lt;i&gt;Trichomonas vaginalis&lt;/i&gt; detected from pregnant women attending Kawo General Hospital, Kaduna, Nigeria

Trichomoniasis is a widespread Sexually Transmitted Infections (STIs) that can have detrimental consequences, including infection of the skin, endometrium, Bartholin glands, and fallopian tubes and ovaries (the adnexa of the uterus). This study was conducted to assess the prevalence of Trichomonas vaginalis (T. vaginalis) and T. vaginalis from vaginal swabs of pregnant patients attending Kawo general hospital in Kaduna state. A total of 250 high vaginal swab samples were randomly taken from consenting pregnant women attending antenatal. Structured questionnaires were used to collect participant demographic and medical data. Samples were analysed by wet mount preparation and T. vaginalis positive samples were confirmed using Polymerase Chains Reaction (PCR). Out of the 250 pregnant women that were examined, 24 (9.6%) were positive for T. vaginalis infection. T. vaginalis was detected more in age group 20-25 (9.4%), and among women of third trimester stage (11.4%). T. vaginalis infection was significantly associated with age, gravidation, education and occupation. Among the 24 pregnant women that tested positive for T. vaginalis infection, only 2 (8.0%) were without symptoms associated with vaginal trichomoniasis, while the remaining 22 (92.0%) presented symptoms such as vaginal odour (20%), itching and discomfort (32%) and greenish yellow vaginal discharge (92%). Molecular analysis confirmed 1 positive case out of the 5 samples randomly analysed. Trichomoniasis transmission to newborn infants could be avoided with better personal cleanliness, screening, and treatment for pregnant women who are affected.