title: Short-Term Exposure to Coal Combustion Waste Has Little Impact on the Skin Microbiome of Adult Spring Peepers (Pseudacris crucifer).
abstract: Disruptions to the microbiome can impact host health as can exposure to environmental contaminants. However, few studies have addressed how environmental contaminants impact the microbiome. We explored this question for frogs that breed in wetlands contaminated with fly ash, a by-product of coal combustion that is enriched in trace elements. We found differences in the bacterial communities among a fly ash-contaminated site and several reference wetlands. We then experimentally assessed the impacts of fly ash on the skin microbiome of adult spring peepers (Pseudacris crucifer). Frogs were exposed to fly ash in the laboratory for 12 h, the duration of a typical breeding event, and the skin microbiome was assessed after 5 days (experiment 1) or after 5 and 15 days (experiment 2). We examined bacterial community structure using 16S rRNA gene amplicon sequencing and metabolite profiles using high-pressure liquid chromatography-mass spectrometry (HPLC-MS). We found little impact as the result of acute exposure to fly ash on the bacterial communities or metabolite profiles in either experiment, suggesting that the bacterial symbiont communities of adults may be relatively resistant to brief contaminant exposure. However, housing frogs in the laboratory altered bacterial community structure in the two experiments, which supports prior research suggesting that environmental source pools are important for maintaining the amphibian skin microbiome. Therefore, for contaminants like fly ash that may alter the potential source pool of symbionts, we think it may be important to explore how contaminants affect the initial assembly of the amphibian skin microbiome in larval amphibians that develop within contaminated sites.
  
  
  
  Animals are hosts to many symbiotic microorganisms, collectively called the microbiome, that play critical roles in host health. Therefore, environmental contaminants that alter the microbiome may impact hosts. Some of the most widespread contaminants, produced worldwide, are derived from the mining, storage, and combustion of coal for energy. Fly ash, for example, is a by-product of coal combustion. It contains compounds such as arsenic, selenium, cadmium, and strontium and is a recognized source of ground and surface water contamination. Here, we experimentally investigated the impacts of short-term fly ash exposure on the skin microbiome of spring peepers, one of many species of amphibian that sometimes breed in open fly ash disposal ponds. This research provides a look into the potential impacts of fly ash on an animal's microbiome and suggests important future directions for research on the effects of environmental contaminants on the microbiome.
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA myrahughey@gmail.com.
authors.forename: Myra C
authors.initials: MC
authors.lastname: Hughey
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Jenifer B
authors.initials: JB
authors.lastname: Walke
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Becker
authors.affiliation: Department of Chemistry, Villanova University, Villanova, Pennsylvania, USA.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Umile
authors.affiliation: Department of Chemistry, Villanova University, Villanova, Pennsylvania, USA.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Burzynski
authors.affiliation: Department of Chemistry, Villanova University, Villanova, Pennsylvania, USA.
authors.forename: Kevin P C
authors.initials: KPC
authors.lastname: Minbiole
authors.affiliation: Department of Chemistry, Villanova University, Villanova, Pennsylvania, USA.
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Iannetta
authors.affiliation: Department of Chemistry, Villanova University, Villanova, Pennsylvania, USA.
authors.forename: Celina N
authors.initials: CN
authors.lastname: Santiago
authors.affiliation: Department of Fish and Wildlife Conservation, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: William A
authors.initials: WA
authors.lastname: Hopkins
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Lisa K
authors.initials: LK
authors.lastname: Belden
mesh_terms: D000818:Animals; D001001:Anura; D001419:Bacteria / Q000145:classification* / Q000235:genetics*; D058448:Biota / Q000187:drug effects*; D016000:Cluster Analysis; D003031:Coal; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004785:Environmental Pollutants / Q000378:metabolism*; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012867:Skin / Q000187:drug effects*
chemical_list: D003031:Coal; D004275:DNA, Ribosomal; D004785:Environmental Pollutants; D012336:RNA, Ribosomal, 16S