ERIC-PCR-based strain-specific detection of phenol-degrading bacteria in activated sludge of wastewater treatment systems

Lett Appl Microbiol. 2009 Oct;49(4):522-8. doi: 10.1111/j.1472-765X.2009.02696.x. Epub 2009 Jul 21.

Abstract

Aims: To evaluate the use of Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR)-derived probes and primers to specifically detect bacterial strains in an activated sludge microbial community.

Methods and results: ERIC-PCR was performed on two phenol-degrading bacterial strains, Arthrobacter nicotianae P1-7 and Klebsiella sp. P8-14. Their amplicons were DIG labelled for use as probes and then hybridized with ERIC-PCR fingerprints. The results showed the distinct band patterns for both bacterial strains. Strain-specific PCR primers were designed based on the sequences of ERIC-PCR bands. The DNA of each of these strains was successfully detected from its mixture with activated sludge DNA, either by using their respective ERIC-PCR-based probes for hybridization or by using species-specific primers for amplification, with higher sensitivity by latter method.

Conclusions: Two phenol-degrading bacterial strains were identified from a mixture of activated sludge by using ERIC-PCR-based methods.

Significance and impact of the study: The study demonstrated that the bacteria, which have important functions in complex wastewater treatment microbial communities, could be specifically detected by using ERIC-PCR fingerprint-based hybridization or amplification.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biodegradation, Environmental
  • DNA Fingerprinting / methods*
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Enterobacteriaceae / genetics
  • Enterobacteriaceae / isolation & purification*
  • Enterobacteriaceae / metabolism
  • Phenol / metabolism*
  • Polymerase Chain Reaction / methods*
  • Repetitive Sequences, Nucleic Acid
  • Sewage / microbiology*
  • Species Specificity
  • Waste Disposal, Fluid

Substances

  • DNA Primers
  • DNA, Bacterial
  • Sewage
  • Phenol