title: Antibiotic discovery throughout the Small World Initiative: A molecular strategy to identify biosynthetic gene clusters involved in antagonistic activity.
abstract: The emergence of bacterial pathogens resistant to all known antibiotics is a global health crisis. Adding to this problem is that major pharmaceutical companies have shifted away from antibiotic discovery due to low profitability. As a result, the pipeline of new antibiotics is essentially dry and many bacteria now resist the effects of most commonly used drugs. To address this global health concern, citizen science through the Small World Initiative (SWI) was formed in 2012. As part of SWI, students isolate bacteria from their local environments, characterize the strains, and assay for antibiotic production. During the 2015 fall semester at Bowling Green State University, students isolated 77 soil-derived bacteria and genetically characterized strains using the 16S rRNA gene, identified strains exhibiting antagonistic activity, and performed an expanded SWI workflow using transposon mutagenesis to identify a biosynthetic gene cluster involved in toxigenic compound production. We identified one mutant with loss of antagonistic activity and through subsequent whole-genome sequencing and linker-mediated PCR identified a 24.9 kb biosynthetic gene locus likely involved in inhibitory activity in that mutant. Further assessment against human pathogens demonstrated the inhibition of Bacillus cereus, Listeria monocytogenes, and methicillin-resistant Staphylococcus aureus in the presence of this compound, thus supporting our molecular strategy as an effective research pipeline for SWI antibiotic discovery and genetic characterization.
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Davis
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Tyler
authors.initials: T
authors.lastname: Sloan
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Krista
authors.initials: K
authors.lastname: Aurelius
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Angela
authors.initials: A
authors.lastname: Barbour
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Elijah
authors.initials: E
authors.lastname: Bodey
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Brigette
authors.initials: B
authors.lastname: Clark
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Celeste
authors.initials: C
authors.lastname: Dennis
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Drown
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Megan
authors.initials: M
authors.lastname: Fleming
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Allison
authors.initials: A
authors.lastname: Humbert
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Glasgo
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Trent
authors.initials: T
authors.lastname: Kerns
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Kelly
authors.initials: K
authors.lastname: Lingro
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: MacKenzie
authors.initials: M
authors.lastname: McMillin
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Meyer
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Breanna
authors.initials: B
authors.lastname: Pope
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: April
authors.initials: A
authors.lastname: Stalevicz
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Brittney
authors.initials: B
authors.lastname: Steffen
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Austin
authors.initials: A
authors.lastname: Steindl
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Carolyn
authors.initials: C
authors.lastname: Williams
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Carmen
authors.initials: C
authors.lastname: Wimberley
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Zenas
authors.affiliation: Department of Biology, Seton Hill University, Greensburg, PA, USA.
authors.forename: Kristen
authors.initials: K
authors.lastname: Butela
authors.affiliation: Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, USA.
authors.forename: Hans
authors.initials: H
authors.lastname: Wildschutte
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000302:isolation & purification*; D001419:Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D053898:Biosynthetic Pathways / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D055808:Drug Discovery / Q000379:methods* / Q000458:organization & administration*; D006801:Humans; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013334:Students; D014481:United States; D014495:Universities
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S
keywords: Small World Initiative; biosynthetic gene cluster; citizen science; pseudomonads