Vaginal Swab Samples Research Articles

PSXIV-9 Effects of Dietary Restriction and One-Carbon Metabolite Supplementation During Early Gestation on the Maternal Gut and Vaginal Microbiota in Cattle

Abstract Maternal diet quality and quantity have significant impacts on both maternal and fetal health and development. The composition and function of the maternal gut microbiome is also significantly influenced by diet. However, little is known about the impact of gestational nutrient restriction on the maternal microbiome during early gestation, which is a critical stage for many developmental processes. Here, we evaluated the effects of dietary restriction and one-carbon metabolite (OCM) supplementation on the ruminal and vaginal microbiota of beef heifers during early pregnancy. For this, 32 beef heifers (approximately 14-month-old) were used in a 2 × 2 factorial experiment with main factors including targeted gain (CON; 0.45 kg/d gain vs. RES; -0.23 kg/d gain), and OCM supplementation (OCM vs. noOCM) (n = 8/treatment). OCM heifers received ruminal-protected methionine (20 g/day) and choline (60 g/day) daily via a fine-ground corn carrier, with weekly intramuscular injections of folate (320 mg) and vitamin B12 (20 mg). The noOCM heifers received only corn carrier and saline injections. Heifers were individually fed and began treatment at breeding (d 0) and remained on treatment until d 63 of gestation. Ruminal fluid and vaginal swab samples were collected at d -2, d 35, and d 63 (at necropsy). Microbiota from samples were assessed using 16S rRNA gene (V3-V4) sequencing. Overall community structure of the ruminal microbiota was affected by gain (PERMANOVA: R2 = 0.04, P < 0.0001), OCM (R2 = 0.013, P = 0.03), time (R2 = 0.16, P < 0.0001) and by their interactions (R2 = 0.08, P = 0.004). The greatest difference in ruminal microbial community structure was between RES and CON heifers (R2 ≥ 0.16, P ≤ 0.003) on d 63. Overall microbial richness (total ASVs) was greater in RES (2,900 vs. 2,737; P = 0.005) but was unaffected by OCM (P = 0.25). Analysis via MaAsLin2 revealed that phyla Firmicutes (31% vs. 22%), Actinobacteriota (0.65% vs. 0.36%) and Chloroflexi were enriched (P < 0.05) in RES heifers. Twenty-five of the 34 differentially abundant genera in the rumen microbiota, including Christensellaceae R7 (5.0% vs 2.6%), Lachnobacterium (2.0% vs. 0.46%), and Butyrivibrio were more relatively abundant in RES; whereas 9 genera including Prevotella (16.5% vs. 31.1%), Ruminococcus (1.6% vs. 2.5%) and Saccharofermentans (0.4% vs. 0.5%) were reduced in RES (P < 0.05) on d 63. Overall vaginal microbiota structure was affected by gain (R2 = 0.02, P = 0.023) and time (R2 = 0.34, P < 0.0001) but not OCM (P >0.05). Community structure of RES was distinct from CON (R2 = 0.16, P = 0.032) on d 63, but microbial community richness and diversity were not influenced by OCM (P > 0.05). On d 63, RES heifers had increased Actinobacteriota (30.5% vs. 17.7%), but reduced Proteobacteriota (6.2% vs. 9.4%) and Fusobacteriota (0.02% vs. 0.94%; P < 0.05). In summary, restricted dietary intake during early gestation induced significant alterations in the ruminal microbiota and its impact also extended to the vaginal microbiota. The composition of these two microbial communities remained largely unaffected by OCM supplement.

188 Characterization of the Vagino-Uterine Microbiota in Pregnant and Non-Pregnant Beef Cattle Using 16S Rrna Gene Sequencing and Culturing

Abstract Emerging evidence suggests that urogenital microbiome-targeted approaches may provide a novel opportunity to reduce the incidence of reproductive failures in cattle. The objectives of this study were to: 1) characterize the vaginal and uterine microbiota between beef cattle that became pregnant via artificial insemination (AI) and those that did not; 2) identify differentially abundant taxa between pregnant and non-pregnant cattle; and 3) characterize the culturable fraction of the vaginal and uterine microbiota using extensive culturing. Vaginal and uterine swab samples were collected from two cohorts of Angus-crossbred cattle: mature cows (n =100) and heifers (n = 72, vaginal swab only) at the time of AI. At 35 days post-AI, pregnancy diagnoses were made via ultrasound. A subset of vaginal and uterine swabs from cows (27 non-pregnant and 31 pregnant) and heifers (26 non-pregnant and 33 pregnant) were selected and processed for microbiota assessment using 16S rRNA gene (V3-V4) sequencing. For culturing, a subset of cryopreserved vaginal and uterine swabs (128) were plated onto three different agar types and cultured, and isolates were identified by near-full length 16S rRNA gene sequencing. Sequencing results revealed 11 taxa that were more abundant in the vaginas of heifers that failed to become pregnant via AI than heifers that did become pregnant. While there was no significant difference in vaginal microbiota community structure (P = 0.21) between pregnant and non-pregnant cows, non-pregnant cows tended to have greater microbial richness and Shannon diversity (P ≤ 0.08) compared with pregnant cows. No differentially abundant taxa were detected in the vaginas of pregnant versus non-pregnant cows. Pregnant cows had a distinct uterine microbiota community structure (R2 = 0.032, P = 0.008) but similar alpha diversity (P > 0.1) compared with non-pregnant cows. Twenty-eight differentially abundant taxa were observed in the uterine microbiota between pregnant and non-pregnant groups, with 11 of them more abundant in pregnant cows. Methanobrevibacter ruminantium (archaea) and Fusobacterium necrophorum were among these 11 positive pregnancy-associated taxa. Although community structure was significantly different between the vagina and uterus, 277 “core taxa” (present in ≥ 80% of all samples) were shared between the two communities. A total of 733 bacterial isolates were recovered from vaginal (512) and uterine (221) swabs under aerobic (n = 363, 83 different species) and anaerobic (n = 370, 69 species) culturing. Potential pathogenic isolates within the Trueperella, Mannheimia and Histophilus genera were most exclusively identified from vaginal swabs. Only two Lactobacillus isolates were recovered. Overall, our results indicate that pregnancy-associated taxonomic signatures are present in the bovine uterine and vaginal microbiota. Among 39 pregnancy status-associated taxa, 41% were unclassified at the genus level, suggesting that shotgun metagenomics and culturomics are needed to identify and characterize the pregnancy-associated taxa in cattle.

Longitudinal analysis of vaginal microbiota during IVF fresh embryo transfer and in early pregnancy.

Non-Lactobacillus-dominated vaginal microbiota has been associated with poor gynecologic health and complications during pregnancy. Lactobacilli and especially Lactobacillus crispatus associate with good reproductive health and dominate the microbiota during healthy pregnancy. We examined whether the composition of vaginal microbiota at the time of fresh embryo transfer (ET) has an impact on the success of in vitro fertilization (IVF) and whether vaginal microbiota changes from IVF-ET to early pregnancy within individuals. Vaginal swab samples were collected from subfertile women at the time of IVF-ET (n = 76) and at the eighth gestational week (n = 21) from those who achieved clinical pregnancy. The microbiota composition was analyzed using 16S rRNA gene amplicon sequencing. L. crispatus was more abundant among the 30 women who achieved clinical pregnancy (46.9% vs. 19.1%, q = 0.039) and the 26 women who had live birth (43.3% vs. 23.1%, q = 0.32) compared to those who did not. Lactobacilli, mainly L. crispatus (76.2%), dominated all early pregnancy samples. Microbiota remained the same, i.e., Lactobacillus-dominated type in 52% (11/21), shifted from one Lactobacillus-dominated type to another in 24% (5/21), or shifted from mixed community to Lactobacillus-dominated type in 24% (5/21) women, but never from Lactobacillus dominance to non-lactobacilli dominance. Our results emphasize the role of L. crispatus in the success of IVF-ET and in early pregnancy. During pregnancy, the microbiota shifted toward L. crispatus dominance even if it was undetectable before pregnancy, indicating that most women hold a reservoir of this beneficial Lactobacillus in their reproductive tract.IMPORTANCEInfertility is a global public health issue which leads many couples to seek fertility treatments, of which in vitro fertilization (IVF) is considered to be the most effective. Still, only about one-third of the women achieve live birth after the first IVF embryo transfer (IVF-ET). Factors affecting embryo implantation are poorly known, but the female reproductive tract microbiota may play a key role. Our study confirms the beneficial role of vaginal lactobacilli, especially Lactobacillus crispatus, in the probability of achieving clinical pregnancy and live birth following IVF-ET. Our findings regarding the intra-individual shift of vaginal microbiota between non-pregnancy and pregnancy states are novel and provide new information about the dynamics of microbiota in the early steps of human reproduction. These findings may help clinicians in their attempts to optimize the conditions for ET by microbiota screening or modulation and timing the ET when the microbiota is the most favorable.

Microbiome alterations in women with pelvic organ prolapse and after anatomical restorative interventions

Pelvic organ prolapse (POP) is a benign gynecological disease in which the pelvic organ descends into the vagina and causes voiding, and defecatory dysfunction, mainly occurs in older women. This study aimed to investigate the vaginal microbiome of POP and associated changes after anatomical restorative pessary or reconstructive pelvic operation. We analyzed the vaginal microbiome using 16S ribosomal RNA gene sequencing and compared the results among patient groups with POP, pessary, and postoperation. We also measured 10 inflammation-related cytokines in vaginal swab samples using multiplex immunoassay. In pelvic organ prolapse, vaginal community status type IV was the most prevalent, which showed a low abundance of Lactobacillus with increased diversity and abundance of anaerobic species. The alpha diversity of species richness was highest in the POP group. The beta diversity distance differed significantly between the three groups (p = 0.001). While human intestinal taxa-associated bacteria were reduced after pessary or operation, vaginitis-associated bacterial composition was altered but vaginal microbiome homeostasis was not improved. IFN-γ, IL-10, IL-12p70, IL-1β, IL-4 and TNF-α levels increased in the pessary group. Therefore, in addition to anatomical restorative treatment, supplementary treatment focusing on the recovery of the vaginal microbiome may be needed to maintain the health of gynecological organs in old age.

Alternative direct-to-amplification cell lysis techniques for forensically relevant non-sperm cells.

While efforts have been made to reduce the pervasive backlog of sexual assault evidence collection kits, the actual laboratory process remains very time-consuming due to the requirement of a differential lysis step before DNA purification, as well as intricate mixture analysis towards the end of the DNA workflow. Recently, an alternative, direct-to-amplification sperm lysis method (using 1 M NaOH) was identified. However, a direct cell lysis method for non-sperm cells has not been identified yet. Thus, the primary objective of this work was to find an alternative method that is quick, inexpensive, and does not require multiple purification steps for the lysis of non-sperm cells in sexual assault samples. In this study, vaginal swab samples were lysed with the control method, prepGEM™, as well as six alternative reagents: alkaline buffer with 25-200 mM NaOH, high-salt stain extraction buffer, modified radioimmunoprecipitation assay (RIPA) buffer, mammalian protein extraction reagent (M-PER™), digitonin buffer, and urea/thiourea buffer. Quantification using Quantifiler® Trio of vaginal and semen lysates revealed that the alkaline (25 mM NaOH) and M-PER™ methods were efficient for the lysis of vaginal epithelial cells without substantial sperm cell lysis. Following quantification, analysis of STR profiles from vaginal lysates revealed that the M-PER™ method showed promising results across all metrics examined, including the percentage of detected STR alleles, mean peak heights, peak height ratio, and interlocus balance. Thus, this method was recommended as an alternative to the traditional differential lysis method for non-sperm cells given its ability to produce amplification-ready lysates without any DNA purification step.

Treponema pallidum Detection at Asymptomatic Oral, Anal, and Vaginal Sites in Adults Reporting Sexual Contact with Persons with Syphilis.

We investigated Treponema pallidum PCR positivity at mucosal sites (oral, anal, and vaginal sites) among adults who had sexual contact with a person with syphilis (syphilis contacts). All syphilis contacts had oral rinse and swab samples collected for testing. Men who have sex with men had anal swab and women had vaginal swab samples collected for testing, regardless of the presence of lesions. Of 407 persons tested, 42 (10%) had early syphilis diagnosed; of those, 19 (45%) tested positive by PCR from any anatomic site and had a positive serologic test. T. pallidum was positive from vaginal samples in 3 women, anal samples in 3 men, and oral cavity samples in 2 women and 3 men, without symptoms at those sites. Three women had no prior syphilis serologic test. T. pallidum detection at asymptomatic mucosal sites suggests early syphilis infections, particularly in cases that would conventionally be staged as latent syphilis of unknown duration.

Trend of HPV Molecular Epidemiology in the Post-Vaccine Era: A 10-Year Study.

Cervical cancer, a major health concern among women worldwide, is closely linked to human papillomavirus (HPV) infection. This study explores the evolving landscape of HPV molecular epidemiology in Taiwan over a decade (2010-2020), where prophylactic HPV vaccination has been implemented since 2007. Analyzing data from 40,561 vaginal swab samples, with 42.0% testing positive for HPV, we reveal shifting trends in HPV genotype distribution and infection patterns. The 12 high-risk genotypes, in order of decreasing percentage, were HPV 52, 58, 16, 18, 51, 56, 39, 59, 33, 31, 45, and 35. The predominant genotypes were HPV 52, 58, and 16, accounting for over 70% of cases annually. The proportions of high-risk and non-high-risk HPV infections varied across age groups. High-risk infections predominated in sexually active individuals aged 30-50 and were mixed-type infections. The composition of high-risk HPV genotypes was generally stable over time; however, HPV31, 33, 39, and 51 significantly decreased over the decade. Of the strains, HPV31 and 33 are shielded by the nonavalent HPV vaccine. However, no reduction was noted for the other seven genotypes. This study offers valuable insights into the post-vaccine HPV epidemiology. Future investigations should delve into HPV vaccines' effects and their implications for cervical cancer prevention strategies. These findings underscore the need for continued surveillance and research to guide effective public health interventions targeting HPV-associated diseases.

Antimicrobial and Phytochemical Activities of Garlic (Allium sativum) on Staphylococcus aureus and Candida albicans Isolated from High Vaginal Swab samples and Female Students with UTI

Studies on antimicrobial and phytochemical activities of garlic on Staphylococcus aureus and Candida albicans isolated from HVS samples and female students with urinary tract infection were investigated. Garlic is used in many homes as condiment or as flavoring agent. The garlic used was purchased from Eke market Agbani in Nkanu West Enugu State Nigeria. Ten Urine and HVS samples were collected from female students using a universal container and swab sticks. Preliminary qualitative phytochemical analysis was done using standard methods. The powder of garlic was macerated in ethanol and chloroform to produce crude extract reconstituted with distilled water to concentrations of 200mg/ml, 100mg/ml, 50mg/ml, 25mg/ml and 12.5mg/ml. The ethanol and chloroform extract of garlic obtained was used to evaluate antimicrobial activity against Staphylococcus aureus and Candida albicans. Also, minimum inhibitory concentration was determined. The garlic extract contained alkaloids, tannins, flavonoids, terpenoids and saponins. The inhibitory effect of ethanol extract started at 200mg/ml with inhibition zones of 2mm against Staphylococcus aureus and Candida albicans. MIC result for garlic extract on Staphylococcus aureus and Candida albicans showed that the extract has inhibitory effect at low concentration of 50mg/ml and inhibition increased at 100mg/ml. Ethanol extract was the most effective extract retarding microbial growth of Staphylococcus aureus and Candida albicans. This study however, justify the use of garlic extract in traditional medicine practice as a therapeutic agent. In cases where possible, the ethanol extracts of garlic should be used at a concentration up to 100mg/ml so as to give a better treatment.

The prevalence of group B streptococcal infection during intrapartum period among high-risk group in labour

Background and objective: Pregnancy associated Group B streptococcal infection is a well-established cause of significant neonatal morbidity and mortality. A cross-sectional study was conducted to determine the prevalence of Group B streptococcal infection among women presenting in labor at different ages and gestational ages and its correlation with different risk factors. Methods: Vaginal swab and urine sample for culture from 150 women having risk factors and being in labor and having risk of developing Group B streptococcal infection from 2nd of November 2020 to 2nd of December 2021. Results: The prevalence of Group B streptococcal infection among risky group women during labor in Maternity Teaching Hospital was 8%. The rate of agreement between the vaginal swab results and urine culture results was 92%. There was no significant association between urine culture results with age, parity and prolonged rupture of membrane. A significant high rate of infected urine in culture was found among women with a very extreme preterm gestational age (p value 0.049), and it was also significant among women with pyrexia (p value 0.001). Conclusion: The prevalence of Group B streptococcal infection among high-risk group of women during labor in Maternity Teaching Hospital of Erbil city was 8%.

Rapid and sensitive Cas12a-based one-step nucleic acid detection with ssDNA-modified crRNA

CRISPR-Cas12a RNA-guided complexes have been developed to facilitate the rapid and sensitive detection of nucleic acids. However, they are limited by the complexity of the operation, risk of carry-over contamination, and degradation of CRISPR RNA (crRNA). In this study, a Cas12a-based single-stranded DNA (ssDNA)-modified crRNA (mD-crRNA)-mediated one-step diagnostic method (CasDOS) was established to overcome these drawbacks. mD-crRNA consisted of wild-type crRNA (Wt-crRNA) with ssDNA extensions at the 3′ and 5′ ends. Compared to Wt-crRNA, mD-crRNA exhibited a 100–1000-fold increase in sensitivity in the one-step assay, reducing the cis-cleavage activity of Cas12a to avoid excessive cleavage of the target DNA in the early stages of the reaction, leading to increased amplification and accumulation of the target amplicons, and improved the speed and intensity of the generated fluorescence signal. The detectability of CasDOS was 16.6 aM for the constructed plasmids of Streptococcus agalactiae (GBS), human papillomavirus type 16 (HPV16), and type 18 (HPV18). In clinical trials, CasDOS achieved 100% accuracy in identifying the known genotypes of the five HPV DNA samples. Moreover, CasDOS showed complete concordance with the qPCR results for GBS detection in ten vaginal or cervical swab samples, with a turnaround time from sampling to results within 30 min. In addition, mD-crRNA remained stable after Ribonuclease R treatment, suggesting that it might be more suitable as a raw material for the CRISPR detection kit. In conclusion, we have developed a universal, rapid, and highly sensitive one-step CRISPR detection assay.

O-259 1.\tComparison of endometrial versus vaginal microbiota in 71 infertile patients

Abstract Study question 1.Mostly, endometrial microbiome probes are acquired by trans-cervical sampling using a “pipelle”. However, it is unclear, whether this approach “contaminates” the endometrial material. Summary answer 1.Vaginal and endometrial microbiome samples are significantly different from each other with respect to (alpha-)diversity showing the endometrial community to be more polymicrobial. What is known already 1. The female genital tract is colonized by a continuum of microbiota, ranging from the typically more diverse, but lower-biomass, bacterial communities in the endometrium to the typically less diverse, but higher-biomass, communities in the vagina (Chen et al., 2017). In reproductive-age women, vaginal bacterial communities are mainly dominated by lactobacilli (Ravel, 2011). By contrast, the composition of the endometrial microbiome has remained less well-characterized. With the more recent introduction of next generation sequencing a more quantitative and comprehensive picture of the composition of endometrial bacterial communities has begun to emerge (Baker et al., 2018). Study design, size, duration 1. The study was conducted at “novum, Zentrum für Reproduktionsmedizin”, Essen / Duisburg, Germany. Of 76 enrolled patients undergoing treatment,73 provided both vaginal swab and endometrial biopsy samples from the same sequencing run between February 2020 and March 2021. All analyses were carried out retrospectively. Information about pregnancy or delivery were extracted from medical records or obtained by telephone or email interviews conducted through June 2022. Participants/materials, setting, methods 1.Amplification of V1-V2 16S rDNA regions was carried out at “dus.ana, Düsseldorf Analytik”, Germany, using the QIAseq 16S screening and regional panels. CLC Microbial Genomic Workbench was used to assess bacterial community composition, alpha diversity, and microbial community structure (PCoA). Pairs of complete corresponding vaginal and endometrial microbiota data were available for 71 patients (two patient was excluded due to too low read counts in the vaginal or the endometrial probe, respectively). Main results and the role of chance 1.Patients exhibiting a mean age of 35 years (range 26 – 42) and a mean body mass index (BMI) of 24,5 (range 18,8 to 38,7). 20 (28,2 %) patients were diagnosed with chronic endometritis (more than 1 plasmacyte per HPF) and 15 (21,1 %) with endometriosis. 40 study participants were found to have undergone past miscarriages, and 11 study participants were found to have given birth to 15 children. 2.Abundance analyses showed differences between vaginal and endometrial samples for the Lactobacillus sp., most pronounced for L. jensenii and L. gasserie. The group of non-lactobacilli was only slightly enlarged in the endometrial group, encompassing mainly G. vaginalis. With regard to diversity (Shannon entropy), both microbiota are significantly different (Kruskal-Wallis test). In addition, distribution of the genital microbial communities illustrated by PCoA indicated that the vaginal samples were clustered together showing variance only in one dimension. However, the endometrial samples were far away from each other and grouped very disperse. Limitations, reasons for caution 1.Group of infertile patients of an IVF center with different underlying clinical diagnosis studied in a retrospective setting. Wider implications of the findings 1.Trans-cervical sampling with a “pipelle” provides an endometrial microbiome which is obviously a more polymicrobial community - with a higher diversity, that may harbor more biological heterogeneity, and hence functionality – than the corresponding vaginal microbiome. Further analyses studying the correlation between sequencing and clinical data are ongoing. Trial registration number 2021-1448

Effect of Lactobacillus delbrueckii subsp. lactis on vaginal radiotherapy for gynecological cancer

The aim of this study was to evaluate the effect of Lactobacillus delbrueckii subsp. lactis (L.del) on vaginal microbiota (VM) dysbiosis and vaginal radiation injury in gynecologic cancer patients. The inhibitory effects of L.del on cervical cancer cells were also studied in vitro. Gynecologic cancer patients receiving radiotherapy were randomized into control and L.del intervention groups. The control group received radiotherapy, while the intervention group received radiotherapy and L.del intervention (1 capsule/day placed into the deep vagina from the first day of radiotherapy until the end of treatment). Vaginal swab samples were collected on the first day pre-treatment and the last day post-treatment. DNA from 54 patients was extracted and assessed by the 16S rRNA sequencing method. Radiotherapy resulted in vaginal microbiome dysbiosis characterized by increased phylogenetic diversity and increased abundance of Brevundimonas, Streptococcus and Prevotella, but a decreased abundance of Lactobacillus. Level 2 vaginal radiation injury was positively associated with the abundance of Brevundimonas and gram-negative non-fermenting bacteria. Administration of L.del attenuated the reduction of Lactobacillus while also inhibiting the abundance of Streptococcus and Prevotella, thereby ameliorating radiotherapy-related vaginal microbiota dysbiosis. CLD inhibited the in vitro proliferation of SiHa cells by altering the expression of BCL2, HPV16-E6, HPV16-E7, IL6, MAP7, BAX, Caspase-3, Caspase-9 and LTF. In conclusion, L. del application can alleviate radiation-induced vaginal dysbiosis and restore Lactobacillus dominance of the vaginal microbiome. Moreover, CLD was found to inhibit cell growth and promote the apoptosis of SiHa cells in vitro. The registration number for this clinical trial is ChiCTR1900021784.

The safety assessment of tampons: illustration of a comprehensive approach for four different products.

We illustrate a comprehensive tampon safety assessment approach that assures products can be used safely. Material biocompatibility, vaginal mucosa assessment, vaginal microbiome evaluation, and in vitro assessment of potential risk of staphylococcal toxic shock syndrome expressed through growth of Staphylococcus aureus (S. aureus) and production of TSST-1 are the four essential portions of the approach. Post-marketing surveillance informs of possible health effects that warrant follow up. The approach meets or exceeds US and international regulatory guidance and is described through the example of four tampon products. Each product is comprised mostly of large molecular weight components (cotton, rayon, polymers) that cannot pass the vaginal mucosa, are widely used across the industry, and replete with a vast body of safety data and a long history of safe use in the category. Quantitative risk assessment of all small molecular weight components assured a sufficient margin of safety supporting their use. Vaginal mucosa assessment confirmed that pressure points, rough edges and/or sharp contact points were absent. A randomized cross-over clinical trial (ClinicalTrials.gov Identifier: NCT03478371) revealed favorable comfort ratings, and few complaints of irritation, burning, stinging, or discomfort upon insertion, wear, and removal. Adverse events were few, mild in severity, self-limited and resolved without treatment. Vaginal microbiota assessment in vitro presented no adverse effect on microbial growth. Culture-independent microbiome analyses from vaginal swab samples obtained during the clinical trial showed no differences attributable to tampon usage, but instead due to statistically significant subject-to-subject variability. Growth of S. aureus and TSST-1 toxin production in the presence of any of the four products in vitro were statistically significantly reduced when compared to medium control alone. The data from the four elements of the comprehensive safety assessment approach illustrated herein confirm that tampons evaluated using this system can be used safely for menstrual protection. A post-marketing surveillance system that monitors and responds to in-market experiences indicated in-use tolerability of the product among consumers, thus confirming the conclusions of the pre-marketing safety assessment.

The microbiota of uterine biopsies, cytobrush and vaginal swabs at artificial insemination in Norwegian red cows

The individual resistance or tolerance against uterine disease in dairy cattle might be related to variations in the uterine tract microbiota. The uterine tract microbiota in dairy cattle is a field of increasing interest. However, its specific taxonomy and functional aspects is under-explored, and information about the microbiota in the endometrium at artificial insemination (AI) is still missing. Although uterine bacteria are likely to be introduced via the vaginal route, it has also been suggested that pathogens can be transferred to the uterus via a hematogenous route. Thus, the microbiota in different layers of the uterine wall may differ. Norwegian Red (NR) is a high fertility breed that also has a high prevalence of subclinical endometritis (SCE), an inflammation of the uterus that has a negative effect on dairy cattle fertility. However, in this breed the negative effect is only moderate, raising the question of whether this may be due to a favorable microbiota. In the present study we investigated the endometrial microbiota in NR at AI by biopsy and cytobrush samples, and comparing this to the vaginal microflora. The second objective was to describe potential differences at both distinct depths of the endometrium, in healthy vs SCE positive NR cows. We sampled 24 lactating and clinically healthy Norwegian red cows in their second heat or more after calving, presented for first AI. First, we obtained a vaginal swab and a cytobrush sample, in addition to a cytotape to investigate the animal's uterine health status with respect to SCE. Secondly, we acquired a biopsy sample from the uterine endometrium. Bacterial DNA from the 16S rRNA gene was extracted and sequenced with Illumina sequencing of the V3–V4 region. Alpha and beta diversity and taxonomic composition was investigated. Our results showed that the microbiota of endometrial biopsies was qualitatively different and more even than that of cytobrush and vaginal swab samples. The cytobrush samples and the vaginal swabs shared a similar taxonomic composition, suggesting that vaginal swabs may suffice to sample the surface-layer uterine microbiota at estrus. The current study gave a description of the microbiota in the healthy and SCE positive NR cows at AI. Our results are valuable as we continue to explore the mechanisms for high fertility in NR, and possible further improvements.

Disposable Device for Bacterial Vaginosis Detection.

Due to the increasing demand for clinical testing of infectious diseases at the point-of-care, the global market claims alternatives for rapid diagnosis tools such as disposable biosensors, avoiding the need for specialized laboratories and skilled personnel. Bacterial vaginosis (BV) is an infectious disease that commonly affects reproductive-age women and predisposes the infection of sexually transmitted diseases. Especially in asymptomatic cases, BV can lead to pelvic inflammatory conditions, postpartum endometritis, and preterm labor. Conventionally, BV diagnosis involves the microscopic analysis of vaginal swab samples; it thus requires highly trained personnel. In response, we report a novel microfluidic paper-based analytical device for BV diagnosis. Sialidase, a biomarker overexpressed in BV, was detected by exploiting an immunosensing mechanism previously discovered by our team. This technology employs a graphene oxide-coated surface as a quencher of fluorescence; the fluorescence of the immunoprobes that do not experiment immunoreactions (antibody-antigen) are deactivated by graphene oxide via non-radiative energy transfer, whereas those immunoprobes undergoing immunoreactions preserve their photoluminescence due to the distance and the low affinity between the immunocomplex and the graphene oxide-coated surface. Our paper-based test was typically carried out within 20 min, and the sample volume was 6 μL. Besides, it was tested with 14 vaginal swabs specimens to discriminate clinical samples of women with normal microbiota from those with BV. Our disposable device represents a new tool to prevent the consequences of BV.

Prevalence and Risk Factors of Trichomonas vaginalis Infection among Female Students of Prince Abubakar Audu University, Nigeria

Objective: This study aimed to determine the prevalence rate and associated risk factors of Trichomonas vaginalis infection among undergraduate female students at Prince Abubakar Audu University, Anyigba, Kogi State, Nigeria.
 Methods: This cross-sectional and institutional-based study involved female students who consented to screening. A total of four hundred (400) high vaginal swab samples were randomly collected from consenting female students and analyzed by wet mount preparation. Demographic and clinical information about the participants was obtained using prepared questionnaires.
 Results: It was observed that the overall prevalence of T. vaginalis infection among the 400 female participants was 170 (42.5%). The age ranges 14–18 and 24-28 had the highest prevalence of 50%, while the age range 19–23 had the least prevalence of 41.2%. The Ebira tribe had the highest prevalence of 60%, while the lowest prevalence was 0.0% among Hausa and others. Islam had the highest prevalence of 66.7%, while Christianity had the lowest prevalence of 40.5%. Students at the 100 level had the highest prevalence of 50%, and those at the 200 level had the least prevalence of 38.5%. There was no significant association between T. vaginalis infection and predisposing factors such as douching, sanitation, and sexual habits. However, the study did find that females who did not use sanitary pads were at a significantly higher risk of infection, while females who did not douche their vagina with antiseptic or deodorant products had a slightly higher risk of infection.
 Conclusion: It is clear from the results of the present study that Prince Abubakar Audu University's undergraduate female students are at a high prevalence of 42.5% for Trichomonas vaginalis infection. The study offers crucial information about the incidence of T. vaginalis infection and the risk factors connected to it among female participants in the study area. The high prevalence of infection highlights the need for increased awareness and education about the infection and its prevention in the study area.

Investigation of SARS-CoV-2 in vaginal secretions of women with coronavirus disease 2019

Abstract Objectives The present study investigates the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the vaginal swabs of female patients diagnosed with coronavirus disease 2019 (COVID-19) based on a positive real-time reverse transcription polymerase chain reaction (RT-PCR) test on a combined throat and nasopharyngeal swab. Methods This study included 48 female patients hospitalized in two tertiary hospitals diagnosed with COVID-19 based on a positive RT-PCR test of the combined throat and nasopharyngeal swab samples, along with clinical and radiological findings. The IBM SPSS software package was used for the statistical analysis of the study data. Results SARS-CoV-2 positivity was detected in only one patient (2.08 %) in the present study from RT-PCR tests of vaginal swab samples. This patient was a 64-year-old, postmenopausal woman who tested positive for SARS-CoV-2 in a RT-PCR test of a vaginal swab sample six days after having tested positive in an RT-PCR test of a combined throat and nasopharyngeal swab. The patient’s partner also tested positive for SARS-CoV-2 in an RT-PCR of a combined throat and nasopharyngeal swab. Conclusions The present study is the first to report the presence of SARS-CoV-2 in vaginal secretions in Türkiye. The authors believe there is a need for studies investigating the presence of SARS-CoV-2 in the semen samples of the male partners of female patients to establish whether the presence of SARS-CoV-2 in vaginal secretions can play a role in the transmission of the virus.

Microbial Isolates And Antibiotic Sensitivity Among Pregnant Women With Abnormal Vaginal Discharge At Nuhu Bamalli Maternity Hospital, Kano

Background: Abnormal vaginal discharge in a pregnant woman increases risk of complications such as abortion, premature rupture of fetal membranes, low birth weight and transmission of HIV to the fetus. Objective: To determine the microbial isolates and antimicrobial sensitivity pattern of abnormal vaginal discharge in pregnant women in Nuhu Bamali maternity hospital Kano. Methods: This was a cross-sectional descriptive study where semi structured interviewer administered questionnaires were employed to a sample of 200 respondents using a simple random sampling technique. High vaginal swab samples were collected from pregnant women with abnormal vaginal discharge at the antenatal clinic. Result: Prevalence of abnormal vaginal discharge among pregnant women in this study was 5.25%. Young maternal age, low parity, lack of western education and unemployment were found to be associated with risk of abnormal vaginal discharge (p˂0.001). The prevalence of positive culture was 83.5% in women with abnormal vaginal discharge; C. albicans (33%), E. coli (23%), S. aureus (17.5%), Klebseilla sp. (7.5%), and Streptococcus sp. (2.5%). Eighty-three percent of microorganism were sensitive to Augmentin, 79% to erythromycin, 62% to Ofloxacin and 41% to gentamicin. Most of the organisms were resistant to amoxicillin and ampicillin. Conclusion: The prevalence of abnormal vaginal discharge in pregnancy is 5.25% with C. albicans and E. coli being the commonest causes. Most of the organisms are sensitive to Augmentin and Erythromycin. Both antibiotics are safe in pregnancy.

Evolution ofanelloviruses from a circovirus-like ancestor through gradual augmentation ofthe jelly-roll capsid protein.

Anelloviruses are highly prevalent in diverse mammals, including humans, but so far have not been linked to any disease and are considered to be part of the 'healthy virome'. These viruses have small circular single-stranded DNA (ssDNA) genomes and encode several proteins with no detectable sequence similarity to proteins of other known viruses. Thus, anelloviruses are the only family of eukaryotic ssDNA viruses currently not included in the realm Monodnaviria. To gain insights into the provenance of these enigmatic viruses, we sequenced more than 250 complete genomes of anelloviruses from nasal and vaginal swab samples of Weddell seal (Leptonychotes weddellii) from Antarctica and a fecal sample of grizzly bear (Ursus arctos horribilis) from the USA and performed a comprehensive family-wide analysis of the signature anellovirus protein ORF1. Using state-of-the-art remote sequence similarity detection approaches and structural modeling with AlphaFold2, we show that ORF1 orthologs from all Anelloviridae genera adopt a jelly-roll fold typical of viral capsid proteins (CPs), establishing an evolutionary link to other eukaryotic ssDNA viruses, specifically, circoviruses. However, unlike CPs of other ssDNA viruses, ORF1 encoded by anelloviruses from different genera display remarkable variation in size, due to insertions into the jelly-roll domain. In particular, the insertion between β-strands H and I forms a projection domain predicted to face away from the capsid surface and function at the interface of virus-host interactions. Consistent with this prediction and supported by recent experimental evidence, the outermost region of the projection domain is a mutational hotspot, where rapid evolution was likely precipitated by the host immune system. Collectively, our findings further expand the known diversity of anelloviruses and explain how anellovirus ORF1 proteins likely diverged from canonical jelly-roll CPs through gradual augmentation of the projection domain. We suggest assigning Anelloviridae to a new phylum, 'Commensaviricota', and including it into the kingdom Shotokuvirae (realm Monodnaviria), alongside Cressdnaviricota and Cossaviricota.

Prevalence of Staphylococcus aureus among Female Patients Attending Rivers State University Teaching Hospital (RSUTH) in Port Harcourt, Nigeria

Objectives: To isolate, identify and determine the prevalence of Staphylococcus aureus among Female Patients attending Rivers State University Teaching Hospital in Port Harcourt using High Vaginal Swab samples.
 Methods: A cross-sectional study was carried out on total of 103 female patients attending the outpatients department and Antenatal clinic of Rivers State University Teaching Hospital. High Vaginal Swab samples were aseptically collected from 55 non-pregnant and 48 six weeks postpartum women. These were then analysed in the Microbiology Laboratory of the Hospital, following standard microbiological procedures. Questions on age, number of sexual partners, knowledge of vaginal infection, symptomatic case, History of previous infection and treatments was responded to by each participant through the completing of questionnaires. Mannitol Salt Agar (MSA) was used for the isolation of the organism and their phenotypic traits were determined based on their cultural and biochemical characteristics.
 Results: The study revealed that 25 (24.3%) consisting of 20 (36.4%) non-pregnant women and 5 (10.4%) Postpartum women were positive for Staphylococcus aureus infection. The prevalence of S. aureus among different groups was statistically significant, (P= .002). The highest percentage of S. aureus isolation (36.4%) was found in non-pregnant population. Prevalence of S. aureus was found highest 10 (18.2%) in 21-25 age group among non-pregnant women and none in 26-30 age group among postpartum women. Data obtained from the questionnaires returned showed that 10 (40%) that tested positive for the organism were asymptomatic as at the time of sample collection, had history of previous reproductive tract infection and had previously undergone treatment. 
 Conclusion: The high rate of vaginal Staphylococcal colonisation observed and the positive correlation between Staphylococus aureus prevalence and age, Child delivery status as well as treatment of previous reproductive tract infection seen in the study suggests the need for awareness on the importance of personal hygiene, proper antibiotic usage and regular screening of females within reproductive age.