Response of 1,2-dichloroethane-adapted microbial communities to ex-situ biostimulation of polluted groundwater.
The microbial community of a groundwater system contaminated by 1,2-dichloroethane (1,2-DCA), a toxic and persistent chlorinated hydrocarbon, has been investigated for its response to biostimulation finalized to 1,2-DCA removal by reductive dehalogenation. The microbial population profile of samples from different wells in the aquifer and from microcosms enriched in the laboratory with different organic electron donors was analyzed by ARISA (Amplified Ribosomal Intergenic Spacer Analysis) and DGGE (Denaturing Gradient Gel Electrophoresis) of 16S rRNA genes. 1,2-DCA was completely removed with release of ethene from most of the microcosms supplemented with lactate, acetate plus formate, while cheese whey supported 1,2-DCA dehalogenation only after a lag period. Microbial species richness deduced from ARISA profiles of the microbial community before and after electron donor amendments indicated that the response of the community to biostimulation was heterogeneous and depended on the well from which groundwater was sampled. Sequencing of 16S rRNA genes separated by DGGE indicated the presence of bacteria previously associated with soils and groundwater polluted by halogenated hydrocarbons or present in consortia active in the removal of these compounds. A PCR assay specific for Desulfitobacterium sp. showed the enrichment of this genus in some of the microcosms. The dehalogenation potential of the microbial community was confirmed by the amplification of dehalogenase-related sequences from the most active microcosms. Cloning and sequencing of PCR products indicated the presence in the metagenome of the bacterial community of a new dehalogenase potentially involved in 1,2-DCA reductive dechlorination.
Marzorati
Massimo
M
DISTAM, Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche, Università degli Studi di Milano, via Celoria 2, 20133 Milan, Italy.
Borin
Sara
S
Brusetti
Lorenzo
L
Daffonchio
Daniele
D
Marsilli
Caterina
C
Carpani
Giovanna
G
de Ferra
Francesca
F
D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology / Q000235:genetics; D002713:Chlorine / Q000737:chemistry / Q000378:metabolism; D003001:Cloning, Molecular; D021901:DNA, Intergenic; D045858:Desulfitobacterium / Q000187:drug effects / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005025:Ethylene Dichlorides / Q000378:metabolism* / Q000494:pharmacology*; D005618:Fresh Water / Q000737:chemistry* / Q000382:microbiology*; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D014871:Water Microbiology; D014874:Water Pollutants, Chemical / Q000378:metabolism* / Q000494:pharmacology; D018508:Water Purification
D021901:DNA, Intergenic; D005025:Ethylene Dichlorides; D012336:RNA, Ribosomal, 16S; D014874:Water Pollutants, Chemical; D002713:Chlorine; C024565:ethylene dichloride; D010088:Oxidoreductases; C112990:tetrachloroethene dehalogenase