1. College of Resources and Environment Engineering, Wuhan University of Technology, Wuhan 430070, China;
2. South China Institute of Environmental Sciences, Ministry of Environmental Protection, Guangzhou 510655, China;
3. The key Laboratory of Water and Air Pollution Control of Guangdong Province, Guangzhou 510655, China;
4. College of Resources and Environment Engineering, Wuhan University of Science and Technology, Wuhan 430081, China;
5. School Environmental Science and Engineering, Zhejiang GongShang University, Hangzhou 310000, China
Identification of the host source of microbial contaminants accurately is the basis for targeted treatment in water. To analyze the feature of pollution in the Pearl River Delta Region, qPCR method was applied to choose the suitable specific bacteroides primer, and culture-method was combined which detected the Fecal coliform and Escherichia coli to assess the water quality and the sources of fecal pollution in target area. The results showed that human-specific marker qHS601F/qBac725R, ruminant-specific marker BacB2-590F/Bac708Rm, porcine-specific marker Bac41F/Bac163R and chicken-specific marker qC160F-HU/qBac265R-HU had higher sensitivity and specificity in the Pearl River Delta region. 14 sampling sites were polluted by human, ruminant and poultry fecal. The intensity of pollution by different source was found to be human > ruminant > poultry. And the concentration of qHS601F/qBac725R had a significant correlationship with Fecal coliform, Escherichia coli and EC23S857F/EC23S857R (P<0.05).It reflects that the major contribute of pollutant source was human.
Byamukama D, Mach R L, Kansiime F, et al. Discrimination efficacy of fecal pollution detection in different aquatic habitats of a high-altitude tropical country, using presumptive coliforms, Escherichia coli, and Clostridium perfringens spores[J]. Applied & Environmental Microbiology, 2005,71(1):65-71.
Zhang C M, Wang X C, Zhou J H, et al. Comparative study on enteric pathogens and their relations with fecal indicator bacteria in urban surface waters[J]. Acta Scientiae Circumstantiae, 2012,32(11):2789-2794.
[6]
Schriewer A, Odagiri M, Wuertz S, et al. Human and animal fecal contamination of community water sources, stored drinking water and hands in rural India measured with validated microbial source tracking assays[J]. American Journal of Tropical Medicine & Hygiene, 2015,93(3):509-16.
[7]
Harwood V J, Brownell M, Wang S, et al. Validation and field testing of library-independent microbial source tracking methods in the Gulf of Mexico[J]. Water Research, 2009,43(19):4812-4819.
[8]
Ahmed W, Yusuf R, Hasan I, et al. Quantitative PCR assay of sewage-associated Bacteroides markers to assess sewage pollution in an urban lake in Dhaka, Bangladesh[J]. Canadian Journal of Microbiology, 2010,56(10):838-845(8).
[9]
Oyafuso Z S, Baxter A E, Hall J E, et al. Widespread detection of human-and ruminant-origin Bacteroidales markers in subtidal waters of the Salish Sea in Washington State[J]. Journal of Water & Health, 2015,13(3):827-37.
[10]
Gawler A H, Beecher J E, Brandão J, et al. Validation of host-specific Bacteriodales 16S rRNA genes as markers to determine the origin of faecal pollution in Atlantic Rim countries of the European Union[J]. Water Research, 2007,41(16):3780-3784.
[11]
Okabe S, Okayama N, Savichtcheva O, et al. Quantification of host-specific Bacteroides-Prevotella 16S rRNA genetic markers for assessment of fecal pollution in freshwater[J]. Applied Microbiology & Biotechnology, 2007,74(4):890-901.
[12]
Villemur R, Imbeau M, Vuong M N, et al. An environmental survey of surface waters using mitochondrial DNA from human, bovine and porcine origin as fecal source tracking markers[J]. Water Research, 2015,69:143-153.
[13]
Kapoor V, Pitkänen T, Ryu H, et al. Distribution of Human-Specific Bacteroidales and Fecal Indicator Bacteria in an Urban Watershed Impacted by Sewage Pollution, Determined Using RNA-and DNA-Based Quantitative PCR Assays[J]. Applied & Environmental Microbiology, 2015,81(1):91-9.
[14]
Mieszkin S, Yala J F, Joubrel R, et al. Phylogenetic analysis of Bacteroidales 16S rRNA gene sequences from human and animal effluents and assessment of ruminant faecal pollution by real-time PCR[J]. Journal of Applied Microbiology, 2010,108(3):974-984.
[15]
Reischer G H, Kasper D C, Steinborn R, et al. Quantitative PCR method for sensitive detection of ruminant fecal pollution in freshwater and evaluation of this method in alpine karstic regions[J]. Applied & Environmental Microbiology, 2006,72(8):5610-5614.
[16]
Bernhard A E, Field K G. A PCR assay To discriminate human and ruminant feces on the basis of host differences in Bacteroides-Prevotella genes encoding 16S rRNA[J]. Applied & Environmental Microbiology, 2000,66(10):4571-4.
[17]
Mieszkin S, Furet J P, Corthier G, et al. Estimation of Pig Fecal Contamination in a River Catchment by Real-Time PCR Using Two Pig-Specific Bacteroidales 16S rRNA Genetic Markers[J]. Applied & Environmental Microbiology, 2009,75(10):3045-3054.
[18]
Kobayashi A, Sano D, Hatori J, et al. Chicken-and duck-associated Bacteroides-Prevotella genetic markers for detecting fecal contamination in environmental water[J]. Applied Microbiology & Biotechnology, 2013,97(16):7427-7437.
[19]
Lu J, Domingo J S, Shanks O C. Identification of chicken-specific fecal microbial sequences using a metagenomic approach[J]. Water Research, 2007,41(16):3561-3574.
[20]
Siefring S, Varma M, Atikovic E, et al. Improved real-time PCR assays for the detection of fecal indicator bacteria in surface waters with different instrument and reagent systems[J]. Journal of Water & Health, 2008,6(6):225-37.
[21]
Chern E C, Siefring S, Paar J, et al. Comparison of quantitative PCR assays for Escherichia coli targeting ribosomal RNA and single copy genes[J]. Letters in Applied Microbiology, 2011, 52(3):298-306.
[22]
Suzuki M T, Taylor L T, Delong E F. Quantitative Analysis of Small-Subunit rRNA Genes in Mixed Microbial Populations via 5'-Nuclease Assays[J]. Applied & Environmental Microbiology, 2000,66(11):4605-14.
[23]
Ballesté E, Bonjoch X, Belanche L A, et al. Molecular indicators used in the development of predictive models for microbial source tracking[J]. Applied & Environmental Microbiology Aem, 2010,76(6):1789-1795.
Schmittgen T D. Real-time quantitative PCR[J]. Methods, 2002, 25(4):383-385.
[27]
Ahmed W, Hughes B, Harwood V J. Current Status of Marker Genes of Bacteroides and Related Taxa for Identifying Sewage Pollution in Environmental Waters[J]. Water, 2016,8(6).
[28]
Shanks O C, Kelty C A, Oshiro R, et al. Data Acceptance Criteria for Standardized Human-Associated Fecal Source Identification Quantitative Real-Time PCR Methods[J]. Applied & Environmental Microbiology, 2016,82(9).
Sokolova E, Aström J, Pettersson T J, et al. Decay of Bacteroidales genetic markers in relation to traditional fecal indicators for water quality modeling of drinking water sources[J]. Environmental Science & Technology, 2011,46(2):892-900.
[32]
Jeanneau L, Solecki O, Wéry N, et al. Relative decay of fecal indicator bacteria and human-associated markers:A microcosm study simulating wastewater input into seawater and freshwater[J]. Environmental Science & Technology, 2012,46(4):2375.
[33]
Li X, Harwood V J, Nayak B, et al. A novel microbial source tracking microarray for pathogen detection and fecal source identification in environmental systems[J]. Environmental Science & Technology, 2015,49(12):7319.
[34]
Rogers S W, Donnelly M, Peed L, et al. Decay of Bacterial Pathogens, Fecal Indicators, and Real-Time Quantitative PCR Genetic Markers in Manure-Amended Soils[J]. Applied & Environmental Microbiology, 2011,77(14):4839-4848.
