Structural Characterization of Vaginal Microbiota in Threatened Abortion Patients with Syndrome of Kidney Deficiency by 16S rRNA Sequencing / 中国实验方剂学杂志

ObjectiveTo investigate the structural features of vaginal microbiota in the early threatened abortion patients with the syndrome of kidney deficiency. MethodThirty-one patients with early threatened abortion of kidney deficiency syndrome （DK-TA group） and 116 women with normal early pregnancy （NP group） attending the First Affiliated Hospital of Guangzhou University of Chinese Medicine from May 2018 to December 2020 were selected. The vaginal secretions were collected for 16S rRNA sequencing， which can reveal the vaginal microbiota composition and differential bacteria between the two groups. ResultThe DK-TA group had higher abundance and diversity of vaginal microbiota than the NP group. The Binary jaccard and unweighted_unifrac distance matrix analysis showed that the similarity， dispersion， abundance， and phylogenetic relationship of vaginal microbiota were significantly different between the two groups. At the phylum level， the DK-TA group had lower relative abundance of Bacteroidetes and Fusobacteria and higher relative abundance of Proteobacteria and Gemmatimonadetes than the NP group. At the genus level， the DK-TA group had lower relative abundance of Sneathia and Bifidobacterium and higher relative abundance of Escherichia-Shigella and Shuttleworthia than the NP group. Linear discriminant analysis Effect Size（LEfSe）revealed that Bacteroidetes， Fusobacteria， and Bifidobacteria were dominant in the NP group and Proteobacteria and Firmicutes in the DK-TA group. The function prediction found that DK-TA was closely associated with 38 functional pathways， including cyclic adenosine monophosphate signaling pathway and regulation of tryptophan channels by inflammatory mediators. In addition， the vaginal differential bacteria between the two groups had significant positive or negative association with the differential metabolic pathways. ConclusionProteobacteria， Gemmatimonadetes， and Bacteroidetes in the vaginal microbiota may be biomarkers for threatened abortion of kidney deficiency syndrome.

Characteristics of vaginal microbiota in pregnant women with premature rupture of membranes and establishment of prediction model / 中华微生物学和免疫学杂志

Objective:To study the characteristics of vaginal microbiota in pregnant women with premature rupture of membranes (PROM) and to establish prediction models for PROM.Methods:This study involved 35 women with preterm premature rupture of membranes (PPROM), 180 with term premature rupture of membranes (TPROM) and 255 term birth cases without premature rupture of membranes (TBWPROM, control group). The V3-V4 hypervariable region sequences in the vaginal samples collected at 16-28 weeks of gestation were detected by 16S rRNA gene next-generation sequencing. The differences in Alpha and Beta diversity, and the attributes and metabolic function prediction of each recognized species among the three groups were analyzed. Subsequently, a random forest model was used to establish the prediction models for PROM using vaginal microbiota species and environmental risk factors.Results:Compared with the control group, the Alpha diversity of the PPROM group was higher (Observed features, P=0.022; Faith_pd index, P=0.024) and Beta diversity was also significantly different (Unweighted UniFrac, P=0.010; Jaccard index, P=0.008). In PPROM cases, Megasphaera genomosp. typeⅠ was significantly increased ( P=0.017) and Lactobacillus mulieris was significantly decreased ( P=0.003). In the patients with TPROM, Megasphaera was significantly increased ( P=0.009) and Lactobacillus mulieris was significantly decreased ( P=0.002). In terms of functional pathways, sulfur oxidation ( P=0.021), methanogenesis from acetate ( P=0.036), L-histidine biosynthesis ( P=0.009), adenosylcobalamin biosynthesis ( P=0.041) and fucose degradation ( P=0.001) were significantly increased in patients with PPROM; L-histidine biosynthesis ( P<0.001) and fucose degradation ( P=0.030) were significantly increased in patients with TPROM. The prediction models were established using the random forest model with vaginal microbiota species and environmental risk factors and the prediction model for PPROM performed well [AUC: 0.739 (95%CI: 0.609-0.869), sensitivity: 0.928, specificity: 0.659, positive predictive value: 0.750, negative predictive value: 0.906], which had a certain reference value. Conclusions:Vaginal microbiota might be related to the development and progression of PROM. Studying the differences in vaginal microbiota might provide a new idea for the prevention and treatment of PROM. Functional prediction provided a direction for further research on the mechanism of PROM. The established prediction model could prevent the occurrence of PPROM and promote maternal and infant health.

Prevalencia de infecciones de transmisión sexual e infecciones vaginales en grupo de mujeres reclusas de la cárcel de Arica

Resumen Introducción: Las infecciones de transmisión sexual (ITS) han incrementado su incidencia universalmente en la última década, incluido Chile. Una de las poblaciones afectadas es la privada de libertad. Objetivo: Evaluar la prevalencia ITS en mujeres del Centro de Detención Preventiva de Arica y Parinacota (Chile) y su asociación con factores biodemográficos. Metodología: En 127 mujeres se realizó un análisis bivariado de los resultados de serología para VHB, VHC, VIH1-2 y VDRL, y un estudio de flujo vaginal convencional microbiológico incluyendo Trichomonas vaginalis, Neisseria gonorrhoeae y Chlamydia trachomatis. Resultados: El 33,1% de las reclusas tuvo al menos una ITS; aquellas menores de 34 años, las consumidoras de drogas y con más de una pareja sexual tuvieron mayor riesgo. Las ITS prevalentes fueron infección por VIH (15,7%) y sífilis (7,9%) asociadas al consumo de drogas y relaciones sexuales antes de 14 años. Trichomonas vaginalis (12,9%) se encontró en mujeres jóvenes con más de una pareja sexual. El 53,2% tuvo un cultivo de flujo vaginal positivo, principalmente con Gardenella vaginalis (32,3%), asociada al mayor número de parejas sexuales y menor tiempo de estadía en reclusión. Candida albicans (11,3%) tuvo mayor prevalencia en mujeres entre 19 y 24 años no heterosexuales. Chlamydia trachomatis, VHB, VHC y N. gonorrhoeae tuvieron prevalencias menores. Conclusión: Existe una alta frecuencia de infección por VIH, sífilis y T vaginalis, predominio de G. vaginalis en aproximadamente un tercio de las mujeres estudiadas y en sobre la mitad de los casos estudiados se comprobó una disbiosis vaginal.

Abstract Background: Sexually transmitted infections (STIs) have increased their incidence worldwide in the last decade, as well as in Chile. One of the affected populations is the deprived of liberty. Aim: To evaluate the STI prevalence in women from the Arica y Parinacota Preventive Detention Center (Chile) and its association with biodemographic factors. Methods: 127 women were studied who underwent a bivariate analysis of the serology results for HBV, HCV, HIV1-2 and VDRL, and a study of conventional microbiological vaginal discharge including Trichomonas vaginalis, Neisseria gonorrhoeae and Chlamydia trachomatis. Results: 33.1% of the inmates had at least one STI, where, women under 34 years old, drug use and more than one sexual partner were at greater risk. The most prevalent STI were HIV infection (15.7%) and syphilis (7.9%) associated with drug use and sexual intercourse before the age of14. Trichomonas vaginalis (12.9%) was identified in young women with more than one sexual partner. 53.2% had a positive culture, mainly with Gardenella vaginalis (32.3%) associated with an increase in sexual partners and a shorter stay in prison. Candida albicans (11.3%) had a higher prevalence in non-heterosexual women between 19 and 24 years old. Chlamydia trachomatis, HBV, HCV and N. gonorrhoeae had lower prevalences. Conclusion: There is a high frequency of HIV infection, syphilis and T. vaginalis, predominance of G. vaginalis in approximately a third of the women studied and about half of the cases studied had vaginal dysbiosis.

Vaginal microbiota characteristics and influencing factors in normal pregnant women / 中华微生物学和免疫学杂志

Objective:To study the characteristics and influencing factors of vaginal microbiota in normal pregnant women.Methods:This study was based on a cohort of pregnant women established in Anqing Municipal Hospital Affiliated to Anhui Medical University from February 2018 to February 2020. Vaginal samples of normal pregnant women who met the inclusion and exclusion criteria were ordered by the gestational weeks at sampling. Five samples were randomly selected from each gestational week group and if the samples were less than five, all samples were included. Sequencing of the V3-V4 region of the 16S rRNA gene was performed. Dominant species were analyzed by MicrobiomeAnalyst. Alpha diversity was measured with Chao1, Observed Features, Shannon diversity, Simpson diversity, Faith_pd and Pielou′s Evenness. The dominant status of Lactobacillus was also described and compared. Multiple linear regression and logistic regression were used to analyze the factors influencing vaginal microbiota. Analysis of variance and Kruskal Wallis test were used for statistical analysis of continuous variables, and Chi-square test and Fisher′s exact test were used for categorical data. The differences were considered statistically significant when the P value was less than 0.05. Results:This study enrolled 91 pregnant women (91 vaginal samples) with an average age of (27.37±3.60) years. There were 18, 56 and 17 vaginal samples collected at the median gestational age of 11.93 weeks (the first trimester), 19.43 weeks (the second trimester) and 38.29 weeks (the third trimester), respectively. The relative abundance of Firmicutes and Lactobacillus was 91.30% and 87.67%, respectively. Lactobacillus iners and Lactobacillus crispatus had a relative abundance of 43.95% and 36.33%, respectively. Moreover, Lactobacillus iners-dominated vaginal microbiota was detected in all trimesters. The number of samples with high relative abundance of Lactobacillus iners gradually decreased with gestational age. Lactobacillus crispatus-dominated vaginal microbiota was found in the second and third trimesters and the number of samples with high relative abundance gradually increased during pregnancy. The Alpha diversity of vaginal microbiota had a decreasing trend during the gestation. There were significant differences in Pielou′s Evenness diversity index of vaginal microbiota between different smoking groups ( P<0.05) and in Shannon diversity index between different drinking groups ( P<0.05). There were significant differences in Chao1, Observed Features and Faith_pd diversity index of vaginal microbiota between pregnant women with different education ( P<0.05) and in Shannon and Simpson diversity index between different income groups ( P<0.05). Conclusions:Vaginal microbiota was dominated by Lactobacillus in normal pregnant women. The dominance of Lactobacillus iners gradually decreased, while that of Lactobacillus crispatus increased during gestation. In normal pregnant women, the Alpha diversity of vaginal microbiota was correlated with smoking, drinking, education and family annual income. Smoking cessation and drinking before pregnancy were related to lower Alpha diversity of vaginal microbiota in pregnant women, while lower education and higher family income were associated with higher Alpha diversity.

Vaginal microbiota transplantation for treatment of bacterial vaginosis: a review / 生物工程学报

Bacterial vaginosis (BV) is a disease caused by vaginal microbiota dysbiosis. The conventional antibiotic treatment can aggravate microbial dysbiosis, alter the acid environment of the vagina and lead to drug resistance, thus shows low cure rate and high recurrence rate. This poses significant physiological and psychological burden to the BV patients. Vaginal microbiota transplantation (VMT) is a novel live biotherapeutic approach. It directly engrafts the whole vaginal microbiota from healthy women to the vaginal tract of patients to rapidly reconstruct the vaginal microbiota environment and restore the vaginal health. This article summarizes the development, present challenges, and future directions of using VMT, with the aim to explore new strategies for treatment of BV and promote the clinical use of VMT.

Vaginal microbiota in pregnancy: Role in induction of labor and seeding the neonate’s microbiota?

Compared to other human microbiota, vaginal microbiota is fairly simple with low bacterial diversity and high relativeabundance of Lactobacillus species. Lactobacillus dominance is even more pronounced during pregnancy. Genetic factors,such as ethnicity, along with environmental, individual and lifestyle factors all have an impact on vaginal microbiotacomposition. The composition of the vaginal microbiota appears to play an important role in pregnancy as recent studieshave linked it to adverse obstetric outcomes such as preterm birth, a leading cause of neonatal morbidity and mortalityworldwide. However, the same vaginal microbiota does not seem to cause the same response in all women, calling forfuture research to fully understand the complex host–microbiota interplay in normal and complicated pregnancies.

Clinical study on the short-term outcome of asymptomatic bacterial vaginosis / 中国实用妇科与产科杂志

OBJECTIVE: To explore the method of management of asymptomatic bacterial vaginosis,and to evaluate the effect of probiotics on asymptomatic bacterial vaginosis and analyze the benefit gained by those patients.METHODS: A multicenter,randomized,double-blind,controlled study was performed with probiotics in test group,and placebo in control group,from september 2013 to september 2016 in 4 hospitals.Forty five patients in test group were treated with vaginal capsule probiotics,and placebo was used in forty-five patients in control group.The follow-up in both groups lasted for 4 weeks.Cure rate,relapse rate and progression rate in both groups were analyzed to evaluate the effect and the benefits gained by patients.RESULTS: Ninty patients were recruited in the study,and seventy-nine patients finished the study and were followed up with complete information.Two weeks after the treatment,in FAS the cure rate in test group was significantly higher than that of the control(44.4% vs. 6.7%,P0.05).In PPS,the cure rate in test group was significantly higher than that of the control(52.5% vs. 2.6%,P0.05).After 2 weeks of treatment,there was no relapsed patient.Four weeks after the treatment,in FAS the cure rate in test group was significantly higher than that of the control(46.7% vs. 8.9%,P0.05).In PPS,the cure rate in test group was significantly higher than that of the control(57.5% vs. 5.1%,P0.05).Four weeks after the treatment,there was no relapsed patient,either.CONCLUSION: Vaginal capsule probiotics can be used to treat asymptomatic BV with high cure rate,less side effect and low relapse rate.

Effect of imblanced vaginal flora on female infertility / 上海交通大学学报（医学版）

Recently the incidence of female infertility is on the rise. The variable and complicated cause of infertility makes it difficult to diagnose in clinical practise. The relationship between vaginal flora and host immune system plays a vital role in female physiological function. Vaginal microbiota dysbiosis may be related to all kinds of infertility, such as tube infertility, endometriosis, anovulatory infertility and idiopathic infertility. In other words, vaginal microbiota dysbiosis may take part in the infertile process. For now the mechanism is far from clear. Some scientists suppose that Lactobacillus deficiency, chronic inflamation and low estrogen level might get involved in this pathophysiological progression, which needs to be studied in the furture.

Infecciones vaginales en un Centro de Salud Familiar de la Región Metropolitana, Chile / Vaginal infections in a Family Health Clinic in the Metropolitan Region, Chile

Background: Vaginal infections are a frequent cause for consultation, but their prevalence and etiology vary in different populations. Objectives: To determine the prevalence and etiologies of vaginal infection in women attending a family health center in the Metropolitan Region of Chile. Methods: The microbiological diagnosis was made by wet mount and Gram stain. Diagnosis of trichomoniasis was performed by wet mount, culture and polymerase chain reaction. Results: 101 women aged 15-54, not selected by signs or symptoms of vaginal infection, 46 of them pregnant were included. In 47 women (46.5%), vaginal infections were diagnosed. An association was observed between age and frequency of vaginal infection. The proportion of infections among pregnant and non-pregnant women was similar. The most frequent infections were bacterial vaginosis (16.8%), vulvovaginal candidiasis (11.9%) and co-infections (6.9%). We found 5.9% of intermediate microbiota cases, 3% of trichomoniasis and 2% of aerobic vaginitis. Symptoms of vaginal infection had poor agreement with microbiological findings. Otherwise physical signs had good agreement with the presence of infection, but low to moderate concordance with a specific etiology. Conclusions: We found a high prevalence of vaginal infections in the study population. It is necessary to improve the definitions and criteria of microbiological diagnosis of co-infections and intermediate microbiota, for them to be diagnosed in the clinical practice. More descriptive questionnaires are recommended to enhance the usefulness of clinical examination.

Introducción: Las infección vaginales constituyen un motivo frecuente de consulta, pero su prevalencia y etiología varían en distintas poblaciones. Objetivos: Determinar la prevalencia y tipos de infección vaginal en mujeres atendidas en un centro de salud familiar de la Región Metropolitana. Métodos: El diagnóstico microbiológico fue efectuado mediante examen microscópico al fresco y tinción de Gram y para tricomoniasis examen al fresco, cultivo y reacción de la polimerasa en cadena. Resultados: Se incluyeron 101 mujeres de 15-54 años, no seleccionadas por signos ó síntomas, 46 de ellas embarazadas. En 47 mujeres (46,5%) se diagnosticaron infecciones vaginales. Se observó asociación entre edad y frecuencia de infección vaginal. La proporción de infecciones entre gestantes y no gestantes fue similar. Las infecciones más frecuentes fueron vaginosis bacteriana (16,8%), candidiasis vulvo-vaginal (11,9%) y co-infecciones (6,9%). Se observó 5,9% casos de microbiota intermedia, 3% de tricomoniasis y 2% de vaginitis aeróbica. Los síntomas de infección vaginal tuvieron mala concordancia con los hallazgos microbiológicos. A su vez, los signos físicos tuvieron buena concordancia con la existencia de infección, pero escasa a moderada concordancia con una etiología específica. Conclusiones: Se encontró alta frecuencia de infecciones vaginales. Es necesario mejorar las definiciones y criterios de diagnóstico microbiológico de las co-infecciones y microbiota intermedia, para ser diagnosticadas en la práctica clínica. Se recomienda emplear cuestionarios más descriptivos para mejorar la utilidad del examen clínico.

Biological control of vaginosis to improve reproductive health.

The human vaginal microbiota plays an important role in the maintenance of a woman’s health, as well as of her partner’s and newborns’. When this predominantly Lactobacillus community is disrupted, decreased in abundance and replaced by different anaerobes, bacterial vaginosis (BV) may occur. BV is associated with ascending infections and obstetrical complications, such as chorioamnionitis and preterm delivery, as well as with urinary tract infections and sexually transmitted infections. In BV the overgrowth of anaerobes produces noxious substances like polyamines and other compounds that trigger the release of pro-inflammatory cytokines interleukin (IL)-1 β and IL-8. BV can profoundly affect, with different mechanisms, all the phases of a woman’s life in relation to reproduction, before pregnancy, during fertilization, through and at the end of pregnancy. BV can directly affect fertility, since an ascending dissemination of the involved species may lead to tubal factor infertility. Moreover, the increased risk of acquiring sexually transmitted diseases contributes to damage to reproductive health. Exogenous strains of lactobacilli have been suggested as a means of re-establishing a normal healthy vaginal flora. Carefully selected probiotic strains can eliminate BV and also exert an antiviral effect, thus reducing viral load and preventing foetal and neonatal infection. The administration of beneficial microorganisms (probiotics) can aid recovery from infection and restore and maintain a healthy vaginal ecosystem, thus improving female health also in relation to reproductive health.

Microbiota cérvico-vaginal de ovelhas mestiças e sua susceptibilidade aos antibióticos / Cervical-vaginal microbiota of crossbred sheep in Petrolina/PE, Brazil, and its susceptibility to antibiotics

A criação de ovinos tem se desenvolvido nas últimas décadas, entretanto ainda são escassas informações sobre a composição e potencial patogênico da microbiota cérvico-vaginal de ovelhas. O presente estudo teve como objetivo conhecer os microrganismos constituintes da microbiota cérvico-vaginal de ovelhas, bem como sua susceptibilidade aos antimicrobianos. Foram realizadas coletas em 60 animais sadios, pertencentes a rebanhos de Petrolina e região. Foi realizado o isolamento bacteriano em ágar sangue e ágar MacConkey, sendo os microrganismos identificados de acordo com características morfológicas, tintoriais e bioquímicas. As amostras foram submetidas ao teste de difusão em disco para determinar o perfil de sensibilidade aos antimicrobianos: sulfametazina, enrofloxacina, doxiciclina, tetraciclina, penicilina, amoxicilina, cefalotina e lincomicina. Foram obtidos 94 isolados, sendo constatada uma maior frequência de Staphylococcus spp. (32,97 por cento), Escherichia coli e Micrococcus spp., sendo observado ainda, isolados de Acinetobacter spp., Shigella spp., Enterobacter spp., Klebsiella spp. e Streptococcus spp. Os isolados apresentaram alta sensibilidade aos antimicrobianos testados sendo observado o menor percentual de sensibilidade para lincomicina. A presença de microrganismos oportunistas de potencial patogênico, na microbiota, como Staphylococcus spp e Escherichia coli, remete a uma análise criteriosa em relação ao diagnóstico de infecções genitais. Os isolados bacterianos obtidos neste estudo são sensíveis à maioria dos grupos de drogas antimicrobianas testadas, demonstrando o potencial de utilização desses princípios ativos, além da disponibilidade de escolha, visto a ausência de multirresistência.

Sheep farming has developed in recent decades; however there is still little information on the composition and pathogenic potential of cervical-vaginal flora of sheep. The purpose of the present study was to determine the main constituents of microorganisms in the cervical-vaginal flora of sheep and their antimicrobial susceptibility. Samples were taken from 60 healthy sheep belonging to herds in the region of Petrolina, Pernambuco. Bacterial isolation was performed on blood agar and MacConkey agar, and the microorganisms were identified according to morphology, Gram staining and biochemical characteristics. The samples were subjected to disk diffusion test for determination of sensitivity to the following antimicrobial drugs: sulfamethazine, enrofloxacin, doxycycline, tetracycline, penicillin, amoxicillin, cephalothin and lincomycin. We obtained 94 isolates and found a higher frequency of Staphylococcus spp., Escherichia coli and Micrococcus spp., and also observed isolates of Acinetobacter spp., Shigella spp., Enterobacter spp., Klebsiella spp. and Streptococcus spp. The isolates were highly sensitive to the antibiotics tested, with the lowest percentage of susceptibility to lincomycin. The presence of opportunistic microorganisms of a potential pathogenic microbiota, such as Staphylococcus spp. and Escherichia coli, refers to a careful analysis in the diagnosis of genital infections. The bacterial isolates obtained in this study are sensitive to most groups of antibiotics tested, demonstrating the potential use of these active ingredients, plus the availability of choice, given the lack of multidrug resistance.

Bacterial vaginosis: clinical, epidemiologic and microbiological features / Bacterial vaginosis: clinical, epidemiologic and microbiological features

Bacterial Vaginosis (BV) is a polymicrobial clinical syndrome, whose etiology has not been fully understood. It occurs in approximately 30% of the women in childbearing age and is the result of the shift of protective resident microorganisms as Lactobacillus spp. by opportunistic pathogenic bacteria such as Gardnerella vaginalis. Patients with BV generally present copious, thin, homogeneous, milky, foul-smelling flow. Vaginal pH is > 4.5 and microscopy reveals bacteria-covered epithelial cells, termed “clue cells”. Around 50% of the patients are asymptomatic and the disease is associated with gynecologic complications, such as cervicitis, salpingitis, endometritis, post-operative infections and pelvic inflammatory disease; and obstetric complications, such as premature rupture of the membranes, preterm deliveries, chorioamniotitis and postpartum endometritis. Although the Nugent´s method is accepted as the “gold standard” for diagnosing BV, Amsel criteria are generally used for diagnosis in clinical practice. The Papanicolaou method is a valid diagnostic option, chiefly when it yields a positive result (mean specificity 95% when compared to gold standard). G. vaginalis has been almost universally recovered from women with BV, plays an important role in the pathogenesis of the disease, besides it may be detected in about 50% of healthy women. According to the literature, first-line drugs for BV treatment worldwide are metronidazole and clindamycin. However, regional studies on G. vaginalis drug susceptibility patterns are needed faced the so fast growing antimicrobial resistance phenomenon.