(492c) Micro-Droplet Co-Cultivation and Characterization of Vaginal Bacteria

The vaginal microbiome (VMB) plays a fundamental role in women’s health and their susceptibility to diseases. For instance, Bacterial vaginosis (BV), the most common genital tract disorder in women of reproductive-age in the United States, is associated with preterm birth, an increased acquisition of HIV, and a higher risk of developing pelvic inflammatory disease. BV is characterized by the replacement of Lactobacillus species by other obligate and facultative anaerobic bacteria, such as Gardnerella vaginalis. Despite its significance, the fundamental questions of how diverse microorganisms in this microbial community interact with one another and with their host still remain largely unanswered. Current approaches for investigating these questions include conventional in vitro microbial cultivation and animal models, of which each has severe limitations such as low throughput and high resource requirement. More recently, microfluidic droplet co-cultivation has emerged as a new technology for dissecting microbial communities in a high-throughput fashion that renders substantial reduction of required reagents, time, and labor. In this work, we employ microfluidic droplet co-cultivation and several downstream analysis methods to recapitulate an antagonistic relationship between G. vaginalis, associated with BV, and Lactobacillus jensenii, a favored lactic acid producing vaginal bacterium. Pools of droplets can be characterized readily by fluorescence in situ hybridization (FISH) or quantitative polymerase chain reaction (qPCR). Furthermore, we show isolation of single droplets and colony formation on agar plates for quantitative analysis based on cell viability. Additionally, multiple displacement amplification (MDA) amplifies DNA content within a single droplet enabling metagenomic analysis. This work demonstrates the foundation of our microdroplet technology and represents the first step of our efforts aiming to elucidate microbial interactions in the human vagina and subsequently to intervene rationally the vaginal microbiome through engineering of probiotics and prebiotics.