Study On The Distribution Of Bacteria Group In Huangpu River In Shanghai

Huangpu River is a branch of the lower reaches of the Yangtze River. It takes its rise from the Tai Lake, runs eastwards through the Dianshan Lake in Qingpu County and then flows to Shanghai. As the mother river of Shanghai, Huangpu River is a multi-functional river with the values of drinking water, shipping, tourism and many other aspects.However, due to high population density of Shanghai, which means the total amount of sewage and the weak pollution control measures to the pollution from many sources such as industry and tourism, the Huangpu River has been a heavy load condition. Therefore,the Huangpu river water environment has been widely concerned by society.In this study, by the method of construction of a 16S rDNA clone library and cultivation of Vibrio cholerae, total plate count and total coli-group, we can understand the distribution of bacteria in the Huangpu River, which can provide evidence for the government to institute environmental improvement policy and disease prevention and control policyThe study includes three aspects as follows.1. Monitoring analysis of Vibrio cholerae in Huangpu River in Shanghai.The sample were collected monthly from sixty-three fixed monitoring points along the Huangpu River from July 2011 to October 2011.The V.cholerae 01 and 0139 strains were isolated and cultured from the aquatic samples, and the strains isolated would be immediately analyzed about their serotype, virulence gene brobes of ctx by PCR method, drug susceptibility by K-B method and their molecular typing by Pulsed field gel electrophoresis (PFGE). The results found that one strain of V.cholerae 0139 was isolated from 252 water samples, with the positive rate of 0.4%. The V.cholerae 0139 strain was found to be non-toxic, and test on susceptibility of the strain to 11 antimicrobial agents show that it was sensitive to ampicillin but resistant to other 10 kinds of antibiotics.Clustering analysis of results from PFGE showed that the strain isolated from Huangpu River and the strain isolated from patient were in different branch, with 76% similarity coefficient. Research shows that Vibrio cholerae was presence in the Huangpu River during the epidemic period of cholera. It’s important to strengthen the surveillance of Vibrio cholerae in aquatic environment to understand it’s pollution situation, so we can get the message in time to make effective measure to prevent and control the epidemic of cholera.2. Detection and analysis of total bacteria count and coliform group in Huangpu River in Shanghai.The samples were collected from sixty-three fixed monitoring points along the Huangpu River from July 2011 to October 2011,and the total bacteria count and coliform group were examined in order to investigate the present status of the microbial pollution of Huangpu River and the spatial-temporal distribution of microbial indicators. The results found that the distribution of total bacterial count and total coliform bacteria were varied in different months (P<0.0001),and the distribution of total bacterial count were varied in different places (P<0.0001). The positive correlation was also found between two types of microbial indicators (P<0.0001). Research shows that microbial pollution in Huangpu River is seriously affected by the time and human activities, and there are significant correlations between two microbial indicators, which show a regular Space-time distribution.3. Study on the bacteria diversity in Huangpu River in Shanghai.In this study, bacteria diversity of samples from three areas of Huangpu River that from Wujing to Lu Pu Bridge, from Lupu Bridge to Yangpu bridge and from Yangpu bridge to Renaissance Island,were analyzed by 16S rDNA sequence analysis from July 2011 to October 2011.The results found that a total of 1135 positive clone was obtained from twelve 16S rDNA clone library in the Huangpu River, which can be divided into 660 OTUs. Phylogenetic analyses placed the 660 OTUs in the following groups of the domain bacteria:Proteobacteria, Actinobacteria, Bacteroidete, Cyanobacteria, Verrucomicrobia, Firmicutes, Acidobacteria, Armatimonadetes, Fusobacteria, Planctomycetes and Unclassified Bacteria.In addition, Proteobacteria, Actinobacteria and Bacteroidete were the predominant phyla,and p-Proteobacteria was the most dominant groups. Neither of the bacterial diversity in all libraries were the same, which present temporal and spatial differences. The results of Blast and the analysis of Phylogeny show that most bacteria were uncultured bacteria, and comparative phylogenetic analysis of the closest relatives of the sequences obtained in the Huangpu River implied the presence of influences of human activities.