title: Genetic changes that correlate with the pine-oil disinfectant-reduced susceptibility mechanism of Staphylococcus aureus.
abstract: OBJECTIVE
  To identify factors associated with the Staphylococcus aureus pine-oil disinfectant-reduced-susceptibility (PD(RS)) mechanism and to describe one possible PD(RS) model.
  
  
  RESULTS
  Comparative genomic sequencing (CGS) and microarray analysis were utilized to detect mutations and transcriptome alterations that occur in a S. aureus PD(RS) mutant. Mutant analysis, antimicrobial gradient plates, growth studies and 3-hydroxy-3-methylglutaryl coenzyme A synthase assays were then performed to confirm the biological consequences of the 'omics' alterations detected in a PD(RS) mutant. CGS uncovered three mutations in a PD(RS) mutant in a(n): alcohol dehydrogenase (adh), catabolite control protein A (ccpA) and an NADPH-flavin oxidoreductase (frp). These mutations lead to increased growth rates; increased transcription of an NAD-dependent D-lactate dehydrogenase gene (ddh); and increased flux through the mevalonate pathway. PD(RS) mutants demonstrated reduced susceptibility to bacitracin and farnesol, and one PD(RS) mutant displayed upregulation of bacA, a bacitracin-resistance gene. Collectively, this evidence demonstrates altered undecaprenol metabolism in PD(RS) mutants.
  
  
  CONCLUSIONS
  The PD(RS) mechanism proposed results from increased catabolic capabilities and increased flux through the mevalonate pathway as well as altered bactoprenol physiology.
  
  
  CONCLUSIONS
  A novel mechanism that bacteria utilize to overcome the killing effects of PD formulations is proposed that is unique from the PD(RS) mechanism of the enterobacteraciae.
authors.affiliation: Department of Biology, New Mexico State University, Las Cruces, NM 88003-8001, USA.
authors.forename: R
authors.initials: R
authors.lastname: Lamichhane-Khadka
authors.forename: J T
authors.initials: JT
authors.lastname: Riordan
authors.forename: A
authors.initials: A
authors.lastname: Delgado
authors.forename: A
authors.initials: A
authors.lastname: Muthaiyan
authors.forename: T D
authors.initials: TD
authors.lastname: Reynolds
authors.forename: B J
authors.initials: BJ
authors.lastname: Wilkinson
authors.forename: J E
authors.initials: JE
authors.lastname: Gustafson
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D004252:DNA Mutational Analysis; D004202:Disinfectants / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial / Q000235:genetics* / Q000502:physiology; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000378:metabolism; D045022:Lactate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D046228:Microarray Analysis; D008826:Microbial Sensitivity Tests; D028223:Pinus / Q000737:chemistry*; D010938:Plant Oils / Q000494:pharmacology*; D012097:Repressor Proteins / Q000235:genetics; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D004202:Disinfectants; D010938:Plant Oils; D012097:Repressor Proteins; C069286:catabolite control proteins, bacteria; D045022:Lactate Dehydrogenases; C071195:D-lactate dehydrogenase; D006904:Hydroxymethylglutaryl-CoA Synthase