Detection of filamentous genus Gordonia in foam samples using genus-specific primers combined with PCR--denaturing gradient gel electrophoresis analysis.
A nested-PCR amplification combined with denaturing gradient gel electrophoresis (PCR-DGGE) approach was used to detect and identify Gordonia populations from wastewater treatment plant foam samples. The PCR-amplified region (position 722-1119) by specifically designed primers G699F and G1096R covered the hypervariable region of the Gordonia 16S rRNA gene sequence. This approach successfully distinguished Gordonia species to the interspecies level. The differential ability of PCR-DGGE analysis was effectively used to separate 12 Gordonia species belonging to different 16S rRNA gene-based phylogenetic lineages into 8 groups. Based on this method, the minimum limit of Gordonia detection was 5 x 10(4) CFU.g -1 in the seeded soil samples. The PCR-DGGE bands obtained were excised and identified by sequence analysis. Gordonia polyisoprenivorans, Gordonia amicalis, DGGE type II Gordonia species, and an uncertain Gordonia species dominated the activated sludge foam samples. Results of this study indicate that the detection and analyses of genus Gordonia within a complex microbial community could be accomplished using the PCR-DGGE approach to a larger extent, with certain limitations. Detection of diverse Gordonia populations in foam samples from wastewater treatment plants revealed the significant role of Gordonia in biological foaming during wastewater treatment. The nested-PCR amplification and DGGE can be used as a diagnostic tool for the early detection of foaming incidents in wastewater treatment plants using Gordonia as indicator organism.
Department of Soil and Environmental Sciences, College of Agriculture and Natural Resources, National Chung Hsing University, 250 Kuo Kuang Road, Taichung, 40227, Taiwan, Republic of China.
Fo-Ting
FT
Shen
Hsuan-Ru
HR
Huang
A B
AB
Arun
Hui-Ling
HL
Lu
Ta-Chen
TC
Lin
P D
PD
Rekha
Chiu-Chung
CC
Young
D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004586:Electrophoresis / Q000379:methods*; D004784:Environmental Monitoring; D033601:Gordonia Bacterium / Q000145:classification / Q000235:genetics* / Q000254:growth & development; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013045:Species Specificity; D014865:Waste Disposal, Fluid; D014871:Water Microbiology
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S