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ABSTRACT Electroactive organisms contribute to metal cycling, pollutant removal, and other redox-driven environmental processes via extracellular electron transfer (EET). Unfortunately, developing genotype-phenotype relationships for electroactive organisms is challenging because EET is necessarily removed from the cell of origin. Microdroplet emulsions, which encapsulate individual cells in aqueous droplets, have been used to study a variety of extracellular phenotypes but have not been applied to investigate EET. Here, we describe the development of a microdroplet emulsion system to sort and enrich EET-capable organisms from complex populations. We validated our system using the model electrogenShewanella oneidensisand described the tooling of a benchtop microfluidic system for oxygen-limited conditions. We demonstrated the enrichment of strains exhibiting electroactive phenotypes from mixed wild-type and EET-deficient populations. As a proof-of-concept application, we collected samples from iron sedimentation in Town Lake (Austin, TX) and subjected them to microdroplet enrichment. We measured an increase in electroactive organisms in the sorted population that was distinct compared to a population growing in bulk culture with Fe(III) as the sole electron acceptor. Finally, two bacterial species not previously shown to be EET-capable,Cronobacter sakazakiiandVagococcus fessus, were further cultured and characterized for electroactivity. Our results demonstrate the utility of microdroplet emulsions for isolating and identifying EET-capable bacteria.IMPORTANCEThis work outlines a new high-throughput method for identifying electroactive bacteria from mixed populations. Electroactive bacteria play key roles in iron trafficking, soil remediation, and pollutant degradation. Many existing methods for identifying electroactive bacteria are coupled to microbial growth and fitness—as a result, the contributions from weak or poor-growing electrogens are often muted. However, extracellular electron transfer (EET) has historically been difficult to study in high-throughput in a mixed population since extracellular reduction is challenging to trace back to the parent cell and there are no suitable fluorescent readouts for EET. Our method circumvents these challenges by utilizing an aqueous microdroplet emulsion wherein a single cell is statistically isolated in a pico- to nano-liter-sized droplet. Then, via fluorescence obtained from copper reduction, the mixed population can be fluorescently sorted and gated by performance. Utilizing our technique, we characterize two previously unrecognized weak electrogensVagococcus fessusandCronobacter sakazakii. 

Abstract There is not a clear understanding of the extent by which naturally occurring reactions can attenuate trichloroethene (TCE) and its daughter products within low permeability zones (LPZs), and addressing this knowledge gap requires advancement of methods to accurately measure in situ volatile chemical concentrations. In this study, a soil coring method that freezes the soil in‐situ (a.k.a., cryogenic coring) was utilized to measure depth‐discrete distributions of TCE and its volatile reaction products through a TCE‐impacted silty clay aquitard, and results were compared with those from adjacent soil cores taken using a conventional coring approach. Vertical concentration profiles of TCE,cis‐1,2‐dichloroethylene (DCE), vinyl chloride (VC), ethane, and methane were all compared between the two coring methods, and results indicate the two coring methods recovered statistically equivalent concentrations of volatiles across most depths of the fine‐grained cohesive clayey soil at the study site. Biotic reductive dechlorination was the dominant TCE reaction pathway at the site; several reduced gasses that are possible markers for abiotic reduction were detected, but their concentrations and intervals of occurrence were not sufficiently consistent to indicate whether they were from abiotic TCE reduction or unrelated biological processes. Overall, cryogenic coring yielded improved recovery of sand lenses compared to conventional coring, but offered no apparent benefits for improved recovery of TCE and its volatile reaction products in the low permeability aquitard material at the site.