title: Inferring community dynamics of organohalide-respiring bacteria in chemostats by covariance of rdhA gene abundance.
abstract: We have developed a novel approach to identifying and quantifying closely related organohalide-respiring bacteria. Our approach made use of the unique genomic associations of specific reductive dehalogenase subunit A encoding genes (rdhA) that exist in known strains of Dehalococcoides mccartyi and Desulfitobacterium and the distinguishing covariance pattern of observed rdhA genes to assign genes to unknown strains. To test this approach, we operated five anaerobic reductively dechlorinating chemostats for 3-4 years with tetrachloroethene and trichloroethene as terminal electron acceptors and lactate/formate as electron donors. The presence and abundance of rdhA genes were determined comprehensively at the community level using a custom-developed Reductive Dehalogenase Chip (RDH Chip) DNA microarray and used to define putative strains of Dehalococcoides mccartyi and Desulfitobacterium sp. This monitoring revealed that stable chemical performance of chemostats was reflected by a stable community of reductively dechlorinating bacteria. However, perturbations introduced by, for example, electron donor limitation or addition of the competing electron acceptor sulfate led to overall changes in the chemostat performance, including incomplete reduction in the chloroethene substrates, and in the population composition of reductively dehalogenating bacteria. Interestingly, there was a high diversity of operationally defined D. mccartyi strains between the chemostats with almost all strains unique to their specific chemostats in spite of similar selective pressure and similar inocula shared between chemostats.
authors.affiliation: Department of Civil and Environmental Engineering, Stanford University, Stanford, CA, USA.
authors.forename: Ian P G
authors.initials: IP
authors.lastname: Marshall
authors.forename: Mohammad F
authors.initials: MF
authors.lastname: Azizian
authors.forename: Lewis
authors.initials: L
authors.lastname: Semprini
authors.forename: Alfred M
authors.initials: AM
authors.lastname: Spormann
mesh_terms: D041862:Chloroflexi / Q000235:genetics* / Q000378:metabolism; D045858:Desulfitobacterium / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial; D020411:Oligonucleotide Array Sequence Analysis; D010088:Oxidoreductases / Q000235:genetics*; D013750:Tetrachloroethylene / Q000378:metabolism; D014241:Trichloroethylene / Q000378:metabolism
chemical_list: D014241:Trichloroethylene; D010088:Oxidoreductases; D013750:Tetrachloroethylene
keywords: DNA microarray; Dehalococcoides mccartyi; Desulfitobacterium; chemostat; reductive dehalogenases