Associations between Vaginal Pathogenic Community and Bacterial Vaginosis in Chinese Reproductive-Age Women

Background

Bacterial vaginosis (BV) is one of the most common urogenital infections among women of reproductive age that represents shifts in microbiota from Lactobacillus spp. to diverse anaerobes. The aim of our study was to evalute the diagnostic values of Gardnerella, Atopobium, Eggerthella, Megasphaera typeI, Leptotrichia/Sneathia and Prevotella, defined as a vaginal pathogenic community for BV and their associations with vaginal pH and Nugent scores.

Methods and Findings

We investigated the vaginal pathogenic bacteria and Lactobacillus spp. with species-specific real-time quantitative PCR (qPCR) in 50 BV-positive and 50 BV-negative Chinese women of reproductive age. Relative to BV-negative subjects, a siginificant decline in Lactobacillus and an obvious increase in bacteria in the vaginal pathogenic community were observed in BV-postive subjects (P<0.05). With the exception of Megasphaera typeI, other vaginal pathogenic bacteria were highly predictable for BV with a better sensitivity and specificity. The vaginal pathogenic community was positively associated with vaginal pH and Nugent scores, while Lactobacillus spp., such as L. iners and L. crispatus was negatively associated with them (P<0.05).

Conclusions

Our data implied that the prevalance of vaginal pathogenic bacteria as well as the depletion of Lactobacillus was highly accurate for BV diagnosis. Vaginal microbiota shifts, especially the overgrowth of the vaginal pathogenic community, showed well diagnostic values in predicting BV. Postive correlations between those vaginal pathogenic bacteria and vaginal pH, Nugent score indicated the vaginal pathogenic community rather than a single vaginal microorganism, was participated in the onset of BV directly.     

RVVC患者阴道分泌物中SP-A的表达及意义

旨在探讨肺表面活性物质相关蛋白A(SP-A)在复发性外阴阴道假丝酵母菌病(RVVC)患者阴道分泌物中的表达及意义。收集RVVC患者、外阴阴道假丝酵母菌患者(VVC)及健康体检者(对照组)阴道分泌物各20例,用酶联免疫吸附测定法(ELISA)测定各组中SP-A蛋白的含量,用逆转录聚合酶链式反应(RT-PCR)方法检测各组中SP-A的基因表达情况。用ELISA方法检测各组中SP-A蛋白的含量发现,VVC组的SP-A明显高于对照组(P0.05),RVVC组的SP-A高于对照组及VVC组(P0.05)。RT-PCR检测结果发现,VVC组的SP-A mRNA的表达水平高于对照组(P0.05),RVVC组的SP-AmR-NA的表达水平明显高于对照组及VVC组(P0.05)。SP-A可能是RVVC患者抗假丝酵母菌的重要物质,其性质或数量的改变可能在RVVC的发病机制中发挥重要作用。     

Understanding Vulvovaginal Candidiasis Through a Community Genomics Approach

Vulvovaginal candidiasis (VVC), predominantly caused by Candida albicans, is one of the most common types of infectious vaginitis. Extensive study has been directed toward understanding host defenses against this infection, and results remain inconclusive. While many have examined the role of innate and cell-mediated immunity, as well as Candida-specific antibodies and the anti-Candida activity of vaginal epithelial cells, little attention has been given to one of the most important players: the vaginal microbiota. Exploring changes in species composition and gene expression within the vaginal community using high-throughput genomic technologies is invaluable to fully understanding Candida pathogenesis and host response to infection. This integrative perspective of pathogenesis, host response and microbial influence are critical to our ability to improve routine gynecologic care and treatment of vaginal infections.     

Prevalence of Candida nivariensis and Candida bracarensis in Vulvovaginal Candidiasis

Candida nivariensis and Candida bracarensis were isolated from patients with vulvovaginal candidiasis (VVC). Candida nivariensis and Candida bracarensis were found in presumptive Candida glabrata isolates, which were identified using the API Candida system. We retrospectively re-examined vaginal presumptive Candida glabrata isolates for Candida nivariensis and Candida bracarensis from January 1, 2003, through December 31, 2012, via detection of the ITS1 region and the 5.8S ribosomal RNA gene. Among 301 presumptive Candida glabrata isolates, 293 isolates were confirmed as C. glabrata (97.34 %), 7 isolates were identified as C. nivariensis (2.33 %) and 1 isolate was identified as C. bracarensis (0.33 %). The C. nivariensis and C. bracarensis isolates were confirmed by sequencing. All C. nivariensis isolates were susceptible to nystatin and susceptible or susceptible dose-dependent to fluconazole, itraconazole, miconazole, and clotrimazole. The C. bracarensis isolate was susceptible to nystatin and the tested azoles. Among the seven patients with VVC caused by C. nivariensis and who were treated with various antifungal agents, only one patient achieved mycological eradication at both the day 7–14 and day 30–35 follow-ups. The C. bracarensis isolate was isolated from a symptomatic pregnant woman; additional data for this patient were unavailable. We conclude that C. nivariensis and C. bracarensis existed in the vaginal samples of patients with VVC. Therapeutic efficacy in the patients with C. nivariensis was poor and inconsistent with the observed in vitro antifungal susceptibility, which requires further study.     

外阴阴道假丝酵母菌病伴发支原体及沙眼衣原体感染的研究

为研究外阴阴道假丝酵母菌病(Vulvovaginal Candidiasis,VVC)患者合并支原体和衣原体感染情况,对86例复发性VVC(recurrent Vulvovaginal Candidiasis,RVVC)患者、219例单纯性VVC患者以及健康妇女200例,分别进行解脲脲原体(Ureaplasma urealyticum,Uu)、人型支原体(mycoplasma hominis,Mh)和沙眼衣原体(Chlamydia trachomatis,CT)检测;所有VVC病例均进行真菌培养。RVVC组和单纯VVC组与对照组之间Uu、Mh及Uu混合Mh阳性率比较,差异均有统计学意义(P0.05)。而三组之间CT和CT混合支原体阳性率比较,均无显著性差异(P0.05)。真菌培养阳性组与阴性组之间,Uu阳性率比较有显著性差异(P0.05),而CT阳性率比较无显著性差异(P0.05)。结果表明,VVC患者合并Uu感染较正常人群明显增高,假丝酵母菌与Uu混合感染可能导致VVC的发生和复发。     

Mouse Strain-Dependent Differences in Estrogen Sensitivity During Vaginal Candidiasis

The animal models available for studying the immune response to genital tract infection require induction of a pseudo estrous state, usually achieved by administration of 17-β-estradiol. In our experimental model of vaginal candidiasis, under pseudo estrus, different strains of mice were used. We observed major differences in the clearance of Candida albicans infection among the different strains, ascribable to differing susceptibility to estradiol treatment. In the early phase of infection CD1, BALB/c, C57BL/6 albino and C57BL/6 mice were colonized to similar levels, while in the late phase of infection, BALB/c mice, which are considered genetically resistant to C. albicans infection, exhibited greater susceptibility to vaginal candidiasis than CD1 and C57BL/6 albino strains of mice. This was because estradiol induced “per se” enlarged and fluid-filled uteri, more pronounced in infected mice and consistently more evident in BALB/c and C57BL/6 mice than in CD1 mice. Unlike CD1, BALB/c and C57BL/6 mice showed a heavy fungal colonization of the uterus, even though C57BL/6 mice apparently cleared C. albicans from the vagina. The presence of C. albicans in the vagina and uterus was accompanied by a heavy bacterial load. Collectively these observations prompted us to carry out a careful analysis of estradiol effects in a mouse model of vaginal infection.     

伊曲康唑治疗复发性外阴阴道念珠菌病38例临床观察

目的了解国产伊曲康唑（商品名“美扶”）治疗复发性外阴阴道念珠菌病的疗效。方法收集38例复发性外阴阴道念珠菌病为治疗组,口服伊曲康唑200mg,1次／d,连续7d,以后每次月经第1天口服伊曲康唑200mg,1次／d,连续6个月经周期停药,而对照组20例则单子硝酸咪康唑栓200mg阴道外用,方法同前。两组完成冲击治疗后1周、3个月、6个月评价疗效。结果1周后治疗组总有效率为92．1％,对照组为90％,两组相比无统计学差异。3个月、6个月后治疗组的复发率分别为3．2％、6．7％,对照组为28．6％、38．5％,两组相比有统计学意义。结论伊曲康唑短程冲击治疗加长期间断给药对复发性外阴阴道念珠菌病的治疗和预防复发效果满意。     

乳杆菌活菌胶囊辅助治疗妊娠期外阴阴道假丝酵母菌病疗效观察

目的探讨乳杆菌活菌胶囊辅助治疗妊娠期外阴阴道假丝酵母菌病疗效观察。方法选取妊娠期外阴阴道假丝酵母菌病患者70例,采用随机数字表将纳入患者分为观察组35例和对照组35例。观察组患者予以乳杆菌活菌胶囊联合制菌素阴道栓治疗,对照组予以单纯的制菌素阴道栓治疗,均连用10d。两组患者停药2周后进行临床疗效观察,并比较治疗后随访3个月的复发率。结果治疗后停药2周,观察组患者的临床总有效率（91．43％）明显优于对照组（71．43％）（χ2=4．63,P〈0．05）;治疗后随访观察3个月,对照组复发11例（44．00％）,观察组复发5例（15．63％）,观察组患者复发率明显低于对照组（χ2=5．60,P〈0．05）。结论乳杆菌活菌胶囊辅助治疗妊娠期外阴阴道假丝酵母菌病疗效好,复发率低,对外阴阴道假丝酵母菌病具有治疗和预防复发的双重功效。     

Genotyping and Persistence of <Emphasis Type="Italic">Candida albicans</Emphasis> from Pregnant Women with Vulvovaginal Candidiasis

Objective

To study Candida albicans genotypes using RAPD and their susceptibility to fluconazole in healthy pregnant women and in vulvovaginal candidiasis (VVC) patients after topical treatment with clotrimazole.

Methods

Vaginal swabs were collected at t = 0 and t = 1 (1 month later) in pregnant women (control group, n = 33), and before (t = 0), at 1 month (t = 1) and at 2 months (t = 2) after clotrimazole treatment in pregnant women with VVC.

Results

Candida albicans was isolated in 30% of healthy pregnant women and 80% of patients with VVC. A high genetic heterogeneity was observed in C. albicans genotypes between individuals. In patients with VVC, topical antifungal treatment with clotrimazole was clinically effective, but only in a 62% C. albicans was eradicated. In patients in which C. albicans was not eradicated, this microorganism persisted for 1 or 2 months after the antifungal treatment. The persistent colonies were not associated with a specific genotype, but they were associated with higher MICs in comparison with colonies isolated from the control group.

Conclusions

Therapy with topical clotrimazole, despite a good clinical outcome, could not eradicate completely C. albicans allowing the persistence of genotypes, with higher MICs to fluconazole. More studies with higher number of patients are needed to validate this preliminary finding.

     

Antifungal Effect of Long Noncoding RNA 9708-1 in the Vulvovaginal Candidiasis Murine Model

Mycopathologia - Vulvovaginal candidiasis (VVC) caused by Candida spp. affects 70–75% of women at least once during their lives. We aim to elucidate the potential mechanism of VVC and...     

Antifungal Susceptibility Profile of Candida Albicans Isolated from Vulvovaginal Candidiasis in Xinjiang Province of China

We investigated the antifungal susceptibility profiles of 207 independent Candida albicans strains isolated from patients with vulvovaginal candidiasis (VVC) in Xinjiang Province of China. Using CLSI M27-A3 and M27-S4 guidelines, anidulafungin and micafungin were the most active drugs against C. albicans showing an MIC₅₀/MIC₉₀ corresponding to 0.016/0.0313 µg/mL, followed by caspofungin (0.25/0.25 µg/mL), posaconazole (0.125/0.5 µg/mL), ravuconazole (0.063/1 µg/mL), itraconazole (0.125/1 µg/mL), amphotericine B (0.5/1 µg/mL), isavuconazole (0.063/2 µg/mL), 5-flucytosine (1/2 µg/mL), voriconazole (0.125/4 µg/mL), and fluconazole (0.5/4 µg/mL). 96.1% (199)–100.0% (207) isolates were sensitive to the three echinocandins tested, amphotericine B and 5-flucytosine. The in vitro activity of triazoles against all isolates tested was variable; itraconazole and voriconazole had reduced the activity to almost half of the isolates (55.1% (114) and 51.2% (106) susceptible, respectively). Fluconazole was active against 76.3% (158) isolates tested. The new triazoles ravuconazole, isavuconazole and posaconazole showed good in vitro potency against 89.9% (186)–95.2% (197) of isolates with the geometric mean MIC (µg/mL) of 0.10, 0.12 and 0.14 µg/mL, respectively. In conclusion, our study indicates that for effective management of systemic candidiasis in Xinjiang Province of China, it is important to determine the susceptibility profiles of isolated C. albicans from patients with VVC.

     

The Relationship of Pregnancy,Vaginal Candidiasis and Glucose Metabolism

In 72 pregnant women with culture-proved vaginal candidiasis, and an equal number of controls matched for parity, size, stage of gestation and age, the intravenous glucose tolerance curves were compared using Silverstone''s method. The results in both study and control groups did not differ and resembled closely those in Silverstone''s series. Half the study group were treated using nystatin alone, while the other half also received tolbutamide. The cure rate was equal in the two groups. Candidiasis in pregnancy does not appear to be related to altered glucose metabolism nor does the addition of this hypoglycemic drug (tolbutamide) improve results. No ill effects, fetal or maternal, were apparent following the use of tolbutamide.     

Increased Diversity of Fungal Flora in the Vagina of Patients with Recurrent Vaginal Candidiasis and Allergic Rhinitis

Recurrent vaginal candidiasis (RVC) is considered to be a hypersensitivity disorder that is associated with allergic rhinitis (AR) in immune deficiencies; however, whether or not the composition of the vaginal fungal flora in patients with AR and RVC is altered and if such alterations in patients with AR are associated with the development of RVC remain unclear. In the present study, a cultivation-independent method with the 18S rRNA gene clone library was used to analyze the diversity and composition of the vaginal fungal flora in patients with AR and RVC and to explore the association. Three fungal phyla (Ascomycotae, 22 out of 28; Basidiomycetes, 5 out of 28; and Oomycetes, 1 out of 28) were identified from groups of healthy volunteers, patients with AR, patients with RVC, and patients with RVC complicated by AR, including 28 phylotypes of fungal flora (10, 15, 17, and 21 phylotypes for each group, respectively). The predominant genera of fungi identified in the vagina included Candida, uncultured fungi, and Dothideomycetes. An increased proportion of Candida albicans accompanied with decreased proportions of Saccharomyces cerevisiae and uncultured fungi was observed in patients with AR or RVC (P?<?0.05). Candida glabrata, Eladia saccula, Trichosporon jirovecii, and Phytophthora spp. occurred simultaneously in the three patient groups. The composition of the fungal communities in the four groups was statistically different (P?<?0.001). The vaginal fungal diversity in patients with AR or RVC was significantly higher compared with healthy volunteers (P?<?0.05). The data revealed an increased diversity and varied composition of the vaginal fungal flora in patients with AR and RVC and indicated that disturbed vaginal fungal flora in patients with AR might be correlated with disease progression in patients with RVC.     

Microbiomes in Canidae

Because of their range expansion across North America, coyotes (Canis latrans) now occur sympatrically with numerous other predator species, including red foxes (Vulpes vulpes). This raises several interesting ecological questions, including if and how sympatry affects the diet and gut microbiomes of coyotes and red foxes. We examined the gut microbiomes of sympatric populations of coyotes and red foxes within two different National Parks in Virginia, USA, that differ in land use, vegetation, and anthropogenic disturbance: Prince William Forest Park (PRWI) and Manassas National Battlefield Park (MANA). From 2012 to 2017, scat samples from PRWI and MANA were collected and analyzed. Polymerase chain reaction (PCR) amplification of a region of the mitochondrial cytochrome‐b gene followed by restriction enzyme digestion of the PCR product was used to determine the origin of each scat sample. Next‐Generation DNA sequencing of a hypervariable 16S rRNA gene region was used to determine gut microbiome information about the scat samples. There was no evidence for a difference between the gut microbiomes of red foxes in either location, or for a difference between the gut microbiomes of red foxes at either location and coyotes at the location with lower human disturbance, PRWI. However, the gut microbiomes of coyotes at the location with higher anthropogenic disturbances, MANA, revealed a marked change from those found in red foxes at either location and from those in coyotes at the location with lower disturbances. The gut microbiomes of coyotes subjected to greater human impact may provide evidence of dysbiosis, indicative of increased physiological stress and reduced health. We discuss our observations in the context of understanding anthropogenic impacts on coyote and red fox interactions. Our results suggest that physiological stress in the form of human disturbance may play an important role in the composition of the gut microbiome of coyotes, which can affect their overall health.     

Genotyping and Drug Resistance Profile of Clinical Isolates of Candida albicans from Vulvovaginal Candidiasis in the Eastern China

Mycopathologia - A total of 244 Candida albicans isolates recovered from vulvovaginal candidiasis (VVC) patients in Suzhou, Eastern China, were investigated. According to CLSI documents M27-A4 and...     

Characteristics of Vaginal Microbiome in Women with Pelvic Inflammatory Disease in Korea

Human vaginal microorganisms play an important role in maintaining good health throughout the human life cycle. An imbalance in the vaginal microbiota is associated with an increased risk of pelvic inflammatory disease (PID). This study aimed to characterize and compare vaginal microbial profiles of premenopausal Korean women with and without PID. 74 Korean premenopausal female vaginal samples were obtained; 33 were from healthy women (a control group) and 41 from PID patients. Vaginal fluid samples were collected from the vaginal wall and posterior cervix and then analyzed by 16S ribosomal ribonucleic acid (rRNA) gene-based amplicon sequencing. Results showed a significant difference between the vaginal microbial communities of the two groups (Jensen-Shannon, p = 0.014; Bray-Curtis, p = 0.009; Generalized UniFrac, p = 0.007; UniFrac, p = 0.008). Lactobacillus accounted for the highest percentage (61.0%) of the control group but was significantly decreased (34.9%) in PID patients; this was the most significant difference among all bacterial communities (p = 0.028, LDA effect size = 5.129). In addition, in the PID patient group, species diversity significantly increased (Simpson, p = 0.07) as the proportion of various pathogens increased evenly, resulting in a polymicrobial infection. Similarly, lactate, which constituted the highest percentage of the organic acids in the control group, was significantly decreased in the PID patient group (p = 0.04). The present study’s findings will help understand PID from the microbiome perspective and are expected to contribute to the development of more efficient PID diagnosis and treatment modalities.