Wet-mount microscopy aerobic vaginitis (AV) diagnostic criteria need phase-contrast microscopy and keen microscopists, and the preservation of saline smears is less common in clinical practice. This research work developed new AV diagnostic criteria that combine Gram stain with clinical features. We enrolled 325 AV patients and 325 controls as a study population to develop new AV diagnostic criteria. Then, an independent group, which included 500 women, was used as a validation population. AV-related microscopic findings on Gram-stained and wet-mount smears from the same participants were compared. The accuracy of bacterial indicators from the two methods was verified by bacterial 16S rRNA V4 sequencing (n = 240). Logistic regression was used to analyse AV-related clinical features. The screened clinical features were combined with Gram-stain microscopic indicators to establish new AV diagnostic criteria. There were no significant differences in the leukocyte counts or the parabasal epitheliocytes (PBC) proportion between the Gram-stain and wet-mount methods (400×). Gram stain (1000×) satisfied the ability to identify bacteria as verified by 16S rRNA sequencing but failed to identify toxic leukocytes. The new criteria included: Lactobacillary grades (LBG) and background flora (Gram stain, 1000×), leukocytes count and PBC proportion (Gram stain, 400×), and clinical features (vaginal pH > 4.5, vagina hyperemia, and yellow discharge). These criteria satisfied the accuracy and reliability for AV diagnosis (Se = 86.79%, Sp = 95.97%, and Kendall’s W value = 0.899) in perspective validation. In summary, we proposed an alternative and valuable AV diagnostic criteria based on the Gram stain, which can make it possible to diagnose common vaginitis like AV, BV, VVC, and mixed infections on the same smear and can be available for artificial intelligence diagnosis in the future.

/pdf/aerobic-vaginitis-diagnosis-criteria-combining-gram-stain-kiell7p6.pdf

Aerobic Vaginitis Diagnosis Criteria Combining Gram Stain with Clinical Features: An Establishment and Prospective Validation Study

Abstract Background Increasing number of studies have demonstrated certain patterns of microbial changes in gynecological diseases; however, the interaction between them remains unclear. To evaluate the consistency or specificity across multiple studies on different gynecological diseases and microbial alterations at different sites of the body (gut and genital tract), we conducted a systematic review and meta-analysis. Methods We searched PubMed, Embase, Web of Science, and Cochrane Library up to December 5, 2022(PROSPERO: CRD42023400205). Eligible studies focused on gynecological diseases in adult women, applied next-generation sequencing on microbiome, and reported outcomes including alpha or beta diversity or relative abundance. The random-effects model on standardized mean difference (SMD) was conducted using the inverse-variance method for alpha diversity indices. Results Of 3327 unique articles, 87 eligible studies were included. Significant decreases were found in gut microbiome of patients versus controls (observed species SMD=-0.35; 95%CI, -0.62 to -0.09; Shannon index SMD=-0.23; 95%CI, -0.40 to -0.06), whereas significant increases were observed in vaginal microbiome (Chao1 SMD = 1.15; 95%CI, 0.74 to 1.56; Shannon index SMD = 0.51; 95%CI, 0.16 to 0.86). Most studies of different diagnostic categories showed no significant differences in beta diversity. Disease specificity was observed, but almost all the changes were only replicated in three studies, except for the increased Aerococcus in bacterial vaginosis (BV). Patients with major gynecological diseases shared the enrichment of Prevotella and depletion of Lactobacillus , and an overlap in microbes was implied between BV, cervical intraepithelial neoplasia, and cervical cancer. Conclusions These findings demonstrated an association between alterations in gut and genital microbiota and gynecological diseases. The most observed results were shared alterations across diseases rather than disease-specific alterations. Therefore, further investigation is required to identify specific biomarkers for diagnosis and treatment in the future. 

Alterations in gut and genital microbiota associated with gynecological diseases: a systematic review and meta-analysis

Abstract Background Aerobic vaginitis (AV) is a vaginal inflammation characterized by disruption of the lactobacillus microbiota and increased counts of different aerobic bacteria. AV may result in severe complications, especially during pregnancy, including preterm delivery, neonatal and maternal infections. This study aimed to determine the prevalence of AV in the third trimester of pregnancy, and the relationship between AV and pregnancy outcomes. Methods A cross-sectional descriptive study included 323 pregnant women attending for routine antenatal care in the Hue University Hospital. Vaginal samples collected at the third trimester of pregnancy were evaluated for AV according to the scoring system of Donders and cultured for identification of predominant bacteria. Pregnancy was followed to its end, and pregnancy outcomes were recorded for both mothers and infants. Results The proportion of pregnant women diagnosed with AV in the third trimester was found to be 15.5%, with the vast majority of the cases (84%) displaying the light AV and 16% the moderate AV. The vaginal cultures in the women with AV revealed most frequently Streptococcus agalactiae (6%), followed by Enterococcus spp (4%), Staphylococcus aureus (4%), and Acinetobacter baumannii (2%). In addition, AV during the last trimester of pregnancy was associated with an increased risk of puerperal sepsis (OR 8.65, 95% CI: 1.41—53.16, p = 0.020) and there was a slightly increased risk for neonatal infections, which was statistically insignificant. Conclusions The proportion of AV is relatively high in Vietnamese pregnant women. Since it is associated with an increased risk of puerperal sepsis, it needs to be diagnosed and treated before delivery. 

/pdf/aerobic-vaginitis-in-the-third-trimester-and-its-impact-on-1vjcik8r.pdf

Aerobic vaginitis in the third trimester and its impact on pregnancy outcomes

Bacterial vaginosis (BV) affects one-third of reproductive age women, increasing the risk of acquiring sexually transmitted infections (STIs) and posing a risk for reproductive health. The current diagnosis with Gram stain (Nugent Score) identifies a transitional stage named partial BV or intermediate microbiota, raising the problem of how to clinically handle it. We retrospectively analyzed cervicovaginal swabs from 985 immunocompetent non-pregnant symptomaticspp. women (vaginal discharge, burning, itching) by Nugent score and qPCR for BV, aerobic or fungal vaginitis, and STIs (Mycoplasmas spp., Chlamydia t., Trichomonas v., and Neisseria g.). Nugent scores 0–3 and 7–10 were confirmed in 99.3% and 89.7% cases, respectively, by qPCR. Among Nugent scores 4–6 (partial BV), qPCR identified 46.1% of BV cases, with 37.3% of cases negative for BV, and only 16.7% of partial BV. Gram staining and qPCR were discordant (p value = 0.0001) mainly in the partial BV. Among the qPCR BV cases, the presence of aerobic vaginitis and STIs was identified, with a significant association (p < 0.0001) between the STIs and partial BV/overt BV. qPCR is more informative and accurate, and its use as an alternative or in combination with Gram staining could help clinicians in having an overview of the complex vaginal microbiota and in the interpretation of partial BV that can correspond to vaginitis and/or STIs.

Vaginal Dysbiosis and Partial Bacterial Vaginosis: The Interpretation of the "Grey Zones" of Clinical Practice.

In the study published by Potter et al. in this issue of Clinical Chemistry (1), 103 genomes under the auspices of either Gardnerella vaginalis or Gardnerella spp. contributed to the National Center for Biotechnology Information database were accessed with the goal of further elucidating subspecies or new species designations (termed genomospecies in the context of in silico analysis). This genus is one of several that can contribute to the dynamic disease entity of bacterial vaginosis. A Canadian group has recently identified differentiating genetic patterns between bacterial vaginosis-, aerobic vaginitis-, and Lactobacillus spp.-dominated vaginal microbiomes (2). Intricate genetic characterization of novel taxa within the Gardnerella genus may identify target organisms for subsequent studies of pathogenesis.

In lieu of the traditional DNA-DNA hybridization standard, Potter et al. used 4 modalities of in silico analysis for subsequent genetic characterization of whole genome assemblies. These included 2 average nucleotide identity (ANI)3 platforms (3), tetranucleotide frequency (4), and average amino acid identity (AAI) (5). The authors used a conservative criterion for assignment of genomospecies (i.e., concordant results derived from ≥2 modalities of characterization). Classifications were supported by ancillary core genome, accessory genome, and metatranscriptome analyses. Using this algorithm, the authors report 9 Gardnerella genomospecies—verifying the taxonomic designation of Gardnerella piotii sp. nov. but suggesting potential conflicts in the taxonomic status of Gardnerella leopoldii sp. nov. and Gardnerella swidsinskii sp. nov. (6).

Past literature has espoused prokaryotic species-level ANI cutoffs of 94% to 96% (3). In a recent publication, Ciufo et al. (7) evaluated this threshold by identifying 335 taxonomic designations for which at least 10 GenBank assemblies were available with ANI alignments above 10% coverage. These assemblies were compared with the submitted type taxon assembly and labeled as concordant (taxonomic agreement) or discordant (taxonomic disagreement). The average ANI for concordant comparisons was 97.1%, …

/pdf/whither-extensive-genomic-based-microbial-taxonomic-revision-54jeqiwjz2.pdf

Whither Extensive Genomic-Based Microbial Taxonomic Revision?

Vaginitis is often diagnosed by microscopy and limited to testing for bacterial vaginosis (BV), vulvovaginal candidiasis, and trichomoniasis. Approximately 10% of vaginal swabs are negative but designated "altered flora" by BV Nugent score, leaving clinicians unsure how to treat patients. Accurate and comprehensive vaginitis diagnostics are needed to direct treatment and reduce risks of recurrent or more severe infections. Vaginal swabs were collected from 93 women (mean age, 23.53 years; range, 18 to 42 years) in a cross-sectional study. Microscopy results for BV and Candida were compared to those from two molecular approaches: (i) a comprehensive quantitative PCR (qPCR) assay, including testing for aerobic vaginitis (AV), Candida, sexually transmitted infections (STI), and BV (Applied Biosystems) with an accompanying BV interpretive algorithm (Coriell Life Sciences), and (ii) microbiome profiling of the 16S rRNA gene (Illumina). Microscopy plus BV Nugent score had 76% overall agreement with the qPCR plus BV interpretive algorithm method (24 positive, 47 negative). OF the nine samples designated altered flora by Nugent, five were categorized BV positive and four were BV negative by the qPCR method. Although BV negative, 3/4 of the latter samples had positive AV targets with one also was STI positive. Microscopic identification of Candida versus that by qPCR had 94% agreement (9 positive, 78 negative). The comprehensive qPCR assay revealed alternative etiologies summarized as 38% BV, 10% AV, 5% Candida, 2% STI, 10% mixed infection (positive targets in multiple panels), and 35% negative for all targets. 16S microbiome analysis confirmed the bacterial qPCR results and identified differentiating patterns between AV, BV, and Lactobacillus-dominated vaginal microbiomes.

Molecular Diagnosis of Vaginitis: Comparing Quantitative PCR and Microbiome Profiling Approaches to Current Microscopy Scoring.

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Molecular Diagnosis of Vaginitis: Comparing Quantitative PCR and Microbiome Profiling Approaches to Current Microscopy Scoring.