Wittman Emma, Yar Neela, De Seta Francesco, Larsen Bryan
College of Osteopathic Medicine, Marian University, Indianapolis, IN 46222, USA.
Department of Medical Sciences, University of Trieste, Institute for Maternal and Child Health-IRCCS, Burlo Garofolo, 34137 Trieste, Italy.
Pathogens. 2021 Mar 3;10(3):289. doi: 10.3390/pathogens10030289.
overgrowth at various mucosal sites is an ongoing and complex clinical concern involving interactions with indigenous microbiota and therapeutic or preventive measures superimposed on the pathogen-microbiome interaction. In this paper we describe the use of quantitative flow cytometry (specific to the cytometer's sample introduction mechanism) to explore the in vitro interaction between , probiotic lactobacilli and a topical vaginal therapeutic. Our central hypothesis was cytometric measurements of co-cultures of yeast and bacteria could provide a useful method for exploring the dynamics of different microbial species in culture, with and without inhibitors. Two commercial products were used as exemplars for this research, a vaginal antimicrobial gel and two species of probiotic lactobacillus intended or oral administration with crystalline bovine lactoferrin to augment the vaginal gel. The cytometer forward channel height parameter distinguished yeast from bacteria in co-culture experiments in the presence of a vaginal therapeutic gel or components of its formulation including EDTA, glycogen, polydextrose as well as the host defense factor, lactoferrin. Flow cytometry showed lactobacilli influenced yeast counts in co-culture, with the technique lending itself to wide-ranging test conditions including organisms, media composition and screening of various antimicrobials. Key findings: The proprietary vaginal gel augmented the effect of lactobacilli, as did EDTA and lactoferrin. Prebiotic compounds also enhanced inhibition by lactobacilli. Propidium iodide (Fluorescence channel 3) discriminated between necrotic and non-necrotic yeast and bacteria in co-cultures under various culture conditions. This research demonstrates the value of flow cytometry to evaluate the population dynamics of yeast and bacteria in co-culture using a proprietary product and its components. We discuss both the limitations of the current study and describe how methods employed here would be transferrable to the investigation of organisms present in defined cultures or at body sites colonized by fungal species and the effects of therapeutics or probiotics on .
各种黏膜部位的过度生长是一个持续且复杂的临床问题，涉及与本地微生物群的相互作用以及叠加在病原体 - 微生物组相互作用之上的治疗或预防措施。在本文中，我们描述了使用定量流式细胞术（特定于细胞仪的样品引入机制）来探索益生菌乳酸杆菌与局部阴道治疗剂之间的体外相互作用。我们的核心假设是，对酵母和细菌共培养物进行细胞计数测量可以提供一种有用的方法，用于探索有或没有抑制剂的培养物中不同微生物物种的动态。本研究使用了两种商业产品作为示例，一种阴道抗菌凝胶和两种旨在口服并与结晶牛乳铁蛋白一起使用以增强阴道凝胶效果的益生菌乳酸杆菌。在存在阴道治疗凝胶或其配方成分（包括乙二胺四乙酸（EDTA）、糖原、聚葡萄糖以及宿主防御因子乳铁蛋白）的共培养实验中，细胞仪的前向通道高度参数可区分酵母和细菌。流式细胞术显示，乳酸杆菌在共培养中影响酵母数量，该技术适用于广泛的测试条件，包括生物体、培养基成分以及各种抗菌剂的筛选。主要发现：专利阴道凝胶增强了乳酸杆菌的作用，EDTA和乳铁蛋白也是如此。益生元化合物也增强了乳酸杆菌的抑制作用。碘化丙啶（荧光通道3）在各种培养条件下的共培养物中区分坏死和非坏死的酵母和细菌。本研究证明了流式细胞术在使用专利产品及其成分评估共培养中酵母和细菌群体动态方面的价值。我们讨论了当前研究的局限性，并描述了此处采用的方法如何可转移用于研究特定培养物中存在的生物体或真菌物种定植的身体部位中的生物体，以及治疗剂或益生菌对其的影响。
