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Droplet-based high-throughput cultivation for accurate screening of antibiotic resistant gut microbes

机译:基于液滴的高通量培养用于精确筛选抗生素抗性肠道微生物

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摘要

Traditional cultivation approaches in microbiology are labor-intensive, low-throughput, and yield biased sampling of environmental microbes due to ecological and evolutionary factors. New strategies are needed for ample representation of rare taxa and slow-growers that are often outcompeted by fast-growers in cultivation experiments. Here we describe a microfluidic platform that anaerobically isolates and cultivates microbial cells in millions of picoliter droplets and automatically sorts them based on colony density to enhance slow-growing organisms. We applied our strategy to a fecal microbiota transplant (FMT) donor stool using multiple growth media, and found significant increase in taxonomic richness and larger representation of rare and clinically relevant taxa among droplet-grown cells compared to conventional plates. Furthermore, screening the FMT donor stool for antibiotic resistance revealed 21 populations that evaded detection in plate-based assessment of antibiotic resistance. Our method improves cultivation-based surveys of diverse microbiomes to gain deeper insights into microbial functioning and lifestyles.
机译:微生物学中的传统栽培方法是由于生态和进化因素,劳动密集型,低通量,并产生了环境微生物的偏见采样。充足的稀有分类群和慢速种植者的充足表示需要新的策略,这些策略通常在培养实验中的快速种植者常用。在这里,我们描述了一种微流体平台,即厌氧分离和培养数百万吡啶液滴中的微生物细胞,并根据菌落密度自动地分类它们以增强生长缓慢的生物体。我们将我们的策略应用于使用多个生长培养基的粪便微生物群移植(FMT)供体粪便,并发现与常规板相比,液滴生长细胞中分类学丰富性和稀有和临床相关征集的较大表示的显着增加。此外,筛选FMT供体粪便用于抗生素抗性,显示出21种群体,其脱落的抗生素抗性评估中的检测。我们的方法改善了各种微生物瘤的培养调查,以获得深入的微生物功能和生活方式的深刻洞察力。

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