Arsenic (As), a toxic element, has impacted life since early Earth. Thus, microorganisms have evolved many As resistance and tolerance mechanisms to improve their survival outcomes given As exposure. We isolated As resistant bacteria from Centralia, PA, the site of an underground coal seam fire that has been burning since 1962. From a 57.4°C soil collected from a vent above the fire, we isolated 25 unique aerobic As resistant bacterial strains spanning seven genera. We examined their diversity, resistance gene content, transformation abilities, inhibitory concentrations, and growth phenotypes. Although As concentrations were low at the time of soil collection (2.58 ppm), isolates had high minimum inhibitory concentrations (MICs) of arsenate and arsenite (>300 mM and 20 mM respectively), and most isolates were capable of arsenate reduction. We screened isolates (PCR and sequencing) using 12 published primer sets for six As resistance genes (AsRGs). Genes encoding arsenate reductase (arsC) and arsenite efflux pumps (arsB, ACR3(2)) were present, and phylogenetic incongruence between 16S rRNA genes and AsRGs provided evidence for horizontal gene transfer. A detailed investigation of differences in isolate growth phenotypes across As concentrations (lag time to exponential growth, maximum growth rate, and maximum OD590) showed a relationship with taxonomy, providing information that could help to predict an isolate’s performance given As exposure in situ. Our results suggest that microbiological management and remediation of environmental As could be informed by taxonomically-linked As tolerance, potential for resistance gene transferability, and the rare biosphere.

23 24Arsenic (As), a toxic element, has impacted life since early Earth. Thus, microorganisms have 25 evolved many As resistance and tolerance mechanisms to improve their survival outcomes given 26 As exposure. We isolated As resistant bacteria from Centralia, PA, the site of an underground 27 coal seam fire that has been burning since 1962. From a 57.4°C soil collected from a vent above 28 the fire, we isolated 25 unique aerobic arsenic resistant bacteria spanning six genera. We 29 examined their diversity, resistance gene content, transformation abilities, inhibitory 30 concentrations, and growth phenotypes. Although As concentrations were low at the time of soil 31PeerJ Preprints | https://doi.org/10.7287/peerj.preprints.2451v2 | CC BY 4.0 Open Access |

23 24Arsenic (As), a toxic element, has impacted life since early Earth. Thus, microorganisms have 25 evolved many As resistance and tolerance mechanisms to improve their survival outcomes given 26 As exposure. We isolated As resistant bacteria from Centralia, PA, the site of an underground 27 coal seam fire that has been burning since 1962. From a 57.4°C soil collected from a vent above 28 the fire, we isolated 25 unique aerobic arsenic resistant bacteria spanning six genera. We 29 examined their diversity, resistance gene content, transformation abilities, inhibitory 30 concentrations, and growth phenotypes. Although As concentrations were low at the time of soil 31PeerJ Preprints | https://doi.org/10.7287/peerj.preprints.2451v2 | CC BY 4.0 Open Access |