Bioremediation Methods and Protocols
Since its first systematic application during the 1970s, bioremediation, or the exploitation of a biological system’s degradative potential to combat toxic pollutants such as heavy metals, polyaromatic hydrocarbons (PAH), cyanides, and radioactive materia
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		    1. Introduction It has been widely recognised that the environment has been polluted by a number of synthetic compounds in the past century, especially after industrialisation. The past few decades have witnessed an increasing interest in bioremediation where living organisms (mainly microorganisms) are used to break down these pollutants in the environment to environmental-friendly compounds. To evaluate the role of microorganisms in the metabolism of certain pollutants in the environment, many techniques need to be employed, including biological and analytical methods, of which stable isotope probing (SIP) is of particular interest (Table 9.1). Stable isotope probing (SIP) relies on the incorporation of stable isotopes (13 C, 15 N) into DNA (1), RNA (2) or phospholipid fatty acid (PLFA) (3–4). Subsequent separation and identification of labelled DNA, RNA or PLFAs offer S.P. Cummings (ed.), Bioremediation, Methods in Molecular Biology 599, DOI 10.1007/978-1-60761-439-5 9, © Humana Press, a part of Springer Science+Business Media, LLC 2010
 
 129
 
 C6 -benzene
 
 C-acetate + perchloroethene
 
 -polychlorinated biphenyls
 
 C-phenanthrene,13 C-pyrene
 
 C-labelled 2,4dichlorophenoxyacetic acid
 
 C-pyrene
 
 C6 -benzene
 
 C6 salicylate; C naphthalene phenanthrene
 
 C-labelled naphthalene and glucose
 
 13
 
 13
 
 13
 
 13
 
 13
 
 13
 
 13
 
 12
 
 13
 
 13
 
 PAH-contaminated soil
 
 C-pyrene
 
 13
 
 Soil
 
 Bioreactor treating PAH-contaminated soil
 
 Gasoline-contaminated groundwater
 
 Bioreactor-treated soil
 
 Agriculture soil
 
 PAH-contaminated soil
 
 Pine tree soil
 
 Pristine river sediment
 
 Coal gasification soil
 
 Habitat
 
 Substrate Acti-
 
 Acidovorax; rangium
 
 Pseudomonas;
 
 Intraspo-
 
 Acidovorax.; Pseudomonas; Ralstonia
 
 16S rRNA
 
 16S rRNA
 
 16S rRNA
 
 16S rRNA
 
 Sphingomonas; uncultivated - and ␥Proteobacteria Azoarcus
 
 16S rRNA
 
 β-Proteobacteria related to Ramlibacter (Comamonadaceae)
 
 16S rRNA; ARHDs1 16S rRNA
 
 Nocar-
 
 16S rRNA
 
 16S rRNA
 
 Acidovorax
 
 Pseudonocardia; Kribbella; dioides; Sphingomonas
 
 Dehalococcoides
 
 Clostridia;
 
 16S rRNA
 
 Uncultivated ␥-Proteobacteria Deltaproteobacteria; nobacteria
 
 Marker genes
 
 Phylogenetic groups identified
 
 Table 9.1 Recent studies using DNA/RNA-SIP for identifying active microorganisms for bioremediation
 
 (continued)
 
 (35)
 
 (34)
 
 (33)
 
 (32)
 
 (31)
 
 (30)
 
 (20)
 
 (29)
 
 (28)
 
 (27)
 
 Reference
 
 130 Chen et al.
 
 C-phenol
 
 C-pentachlorophenol
 
 C-phenol
 
 CH3 Cl CH3 Br
 
 C6 -benzene; 13 C7 -toluene
 
 C-benzoic acid
 
 13
 
 13
 
 13
 
 13
 
 13
 
 13
 
 Agriculture soil
 
 Garden soil
 
 Soil Soil
 
 Agriculture soil
 
 Pristine grassland soil
 
 Activated sludge
 
 Marine sediment or contaminated sediment
 
 Habitat
 
 1 Aromatic ring hydroxylating dioxygenase – Not available
 
 13
 
 C7 -benzoate
 
 13
 
 Substrate
 
 Table 9.1 (Continued)
 
 Burkholderia;
 
 Sphin-
 
 Burkholderia
 
 Arthrobacter; Hydrogenophaga; Pseudomonas; Rhodococcus
 
 Hyphomicrobium; Aminobacter Burkholderia
 
 Kocuria; Staphylococcus; Pseudomonas
 
 Pseudomonas; gomonas
 
 Acidovorax
 
 –
 
 Phylogenetic groups identified
 
 16S rRNA
 
 16S rRNA; bphA; todC
 
 cmuA 16S rRNA; cmuA
 
 16S rRNA
 
 16S rRNA
 
 16S rRNA
 
 nosZ
 
 Ma		
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