Detection of nitrifying bacteria in activated sludge by fluorescent in situ hybridization and fluorescence spectrometry |
| |
Authors: | Kim In S Ivanov Volodymyr N |
| |
Institution: | (1) Department of Environmental Science and Engineering, Kwangju Institute of Science and Technology, 1 Oryong-dong, Buk-gu, Kwangju, 500-712, Republic of Korea;(2) School of Civil & Structural Engineering, Nanyang Technological University, Block N1, Nanyang Avenue, Singapore, 639798 |
| |
Abstract: | 16S rRNA-targeted oligonucleotide probes for eubacteria (EUB338), ammonium-oxidizing bacteria (Nsm156) and nitrite-oxidizing bacteria (Nb1000) were used for the rapid detection of nitrifying bacteria in the activated sludge of a pilot nitrifying reactor by whole-cell, fluorescent in situ hybridization (FISH). Emission scanning and synchronous scanning fluorescence spectrometry were used to measure the hybridization. The binding of the probes at a temperature significantly lower than the melting temperature of the hybrids was conventionally considered as non-specific. Total binding of the probes at a temperature significantly higher than the melting temperature of the hybrids was conventionally considered as the sum of non-specific and specific binding (hybridization). Non-specific binding of the oligonucleotide probes with a biomass of activated sludge was 37% of the total binding of the EUB338 probe, 54% of the total binding of the Nsm156 probe, and 69% of the total binding of the Nb1000 probe. The ratio of the specific binding of the Nsm156 and Nb1000 probes was 2.3:1. The ratio of the numbers of ammonium-oxidizing bacteria to nitrite-oxidizing bacteria, determined by microbiological methods, was 2.4:1. Measuring fluorescent in situ hybridization by fluorescence spectrometry appears to be a practical tool for monitoring the microbial communities that contain nitrifying bacteria. However, a method that accounts for the non-specific binding of the probes more easily and reliably should be developed for practical application. |
| |
Keywords: | Fluorescent in situ hybridization fluorescence spectrometry nitrifying bacteria oligonucleotide probes |
本文献已被 SpringerLink 等数据库收录! |
|