Cell-free supernatants of probiotic consortia impede hyphal formation and disperse biofilms of vulvovaginal candidiasis causing Candida in an ex-vivo model

Sudaarsan Aruna Senthil Kumar, Dhesiga Krishnan, Sowndarya Jothipandiyan, Ramyadevi Durai, B. Narayanan Vedha Hari, Paramasivam Nithyanand
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Abstract

Vulvovaginal candidiasis is the second most common vaginal infection caused by drug-resistant Candida species that affects about 70–75% of reproductive age group women across the globe. As current-day antifungal drugs are ineffective against the biofilms formed by the drug-resistant Candida strains, several natural compounds and antagonistic microbes are being explored as alternative antifungal agents. In the present study, we investigated the anti-biofilm activity of Cell-Free Supernatant (CFS) extracted from the commercially available probiotics VSL-3 against the biofilms of Candida species and also evaluated their efficacy in curbing the yeast-to-hyphal transition. Various methodologies like crystal violet staining and scanning electron microscopy were used to study the effect of CFS against the biofilms formed by the species. The ability of CFS to interfere with yeast to hyphal transition in Candida was studied by colony morphology assay and visually confirmed with phase contrast microscopy. The potential of the CFS of the probiotics was also evaluated using goat buccal tissue, a novel ex-vivo model that mimics the vaginal environment. Moreover, the supernatant extracted from VSL-3 had the ability to down-regulate the expression of virulence genes of Candida from the biofilm formed over the ex-vivo model. These results emphasize the anti-fungal and anti-infective properties of the CFS of VSL-3 against drug-resistant Candida strains causing vulvovaginal candidiasis.

Abstract Image

在体外模型中,益生菌联合体的无细胞上清液可阻碍外阴阴道念珠菌病致病念珠菌的菌丝形成并驱散其生物膜
外阴阴道念珠菌病是由耐药念珠菌引起的第二大常见阴道感染,全球约有 70-75% 的育龄妇女患有该病。由于目前的抗真菌药物对耐药念珠菌菌株形成的生物膜无效,人们正在探索一些天然化合物和拮抗微生物作为替代抗真菌药物。在本研究中,我们研究了从市售益生菌 VSL-3 中提取的无细胞上清液(CFS)对白色念珠菌生物膜的抗生物膜活性,并评估了其抑制酵母菌向真菌转化的功效。研究采用了水晶紫染色和扫描电子显微镜等多种方法来研究 CFS 对念珠菌形成的生物膜的影响。通过菌落形态测定法研究了 CFS 干扰念珠菌从酵母向真菌转化的能力,并用相衬显微镜进行了直观确认。此外,还使用山羊颊组织(一种模拟阴道环境的新型体外模型)对益生菌 CFS 的潜力进行了评估。此外,从 VSL-3 提取的上清液还能下调体内外模型上形成的生物膜中念珠菌毒力基因的表达。这些结果表明,VSL-3 的 CFS 对引起外阴阴道念珠菌病的耐药念珠菌菌株具有抗真菌和抗感染特性。
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