title: In situ exposure to low herbicide concentrations affects microbial population composition and catabolic gene frequency in an aerobic shallow aquifer.
abstract: The aim of this study was to evaluate how the in situ exposure of a Danish subsurface aquifer to phenoxy acid herbicides at low concentrations (<40 micro g l(-1)) changes the microbial community composition. Sediment and groundwater samples were collected inside and outside the herbicide-exposed area and were analyzed for the presence of general microbial populations, Pseudomonas bacteria, and specific phenoxy acid degraders. Both culture-dependent and culture-independent methods were applied. The abundance of microbial phenoxy acid degraders (10(0) to 10(4) g(-1) sediment) was determined by most probable number assays, and their presence was only detected in herbicide-exposed sediments. Similarly, PCR analysis showed that the 2,4-dichlorophenoxyacetic acid degradation pathway genes tfdA and tfdB (10(2) to 10(3) gene copies g(-1) sediment) were only detected in sediments from contaminated areas of the aquifer. PCR-restriction fragment length polymorphism measurements demonstrated the presence of different populations of tfd genes, suggesting that the in situ herbicide degradation was caused by the activity of a heterogeneous population of phenoxy acid degraders. The number of Pseudomonas bacteria measured by either PCR or plating on selective agar media was higher in sediments subjected to high levels of phenoxy acid. Furthermore, high numbers of CFU compared to direct counting of 4',6-diamidino-2-phenylindole-stained cells in the microscope suggested an increased culturability of the indigenous microbial communities from acclimated sediments. The findings of this study demonstrate that continuous exposure to low herbicide concentrations can markedly change the bacterial community composition of a subsurface aquifer.
authors.affiliation: Department of Geochemistry, Geological Survey of Denmark and Greenland, Øster Voldgade 10, DK-1350 Copenhagen K, Denmark.
authors.forename: Julia R
authors.initials: JR
authors.lastname: de Lipthay
authors.forename: Nina
authors.initials: N
authors.lastname: Tuxen
authors.forename: Kaare
authors.initials: K
authors.lastname: Johnsen
authors.forename: Lars H
authors.initials: LH
authors.lastname: Hansen
authors.forename: Hans-Jørgen
authors.initials: HJ
authors.lastname: Albrechtsen
authors.forename: Poul L
authors.initials: PL
authors.lastname: Bjerg
authors.forename: Jens
authors.initials: J
authors.lastname: Aamand
mesh_terms: D015084:2,4-Dichlorophenoxyacetic Acid / Q000378:metabolism; D000332:Aerobiosis; D001419:Bacteria / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D049308:Dioxygenases; D017753:Ecosystem; D005618:Fresh Water / Q000737:chemistry / Q000382:microbiology*; D019015:Geologic Sediments / Q000382:microbiology; D006540:Herbicides / Q000494:pharmacology*; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D010105:Oxygenases / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D006540:Herbicides; D012336:RNA, Ribosomal, 16S; D015084:2,4-Dichlorophenoxyacetic Acid; D006899:Mixed Function Oxygenases; D010105:Oxygenases; D049308:Dioxygenases; C044173:chlorocatechol 1,2-dioxygenase; C084087:2,4-dichlorophenoxyacetate-alpha-ketoglutarate dioxygenase