Studies On Pathogenic Bacteria Distribution In Qinhuangdao Costal Region By Quantitative Polymerase Chain Reaction

This study systematically investigated five representative marine pathogens:Escherichia coli,Bacillus cereus,Vibrio parahemolyticus,Salmonella,and Staphylococcus aureus using quantitative PCR strategy to identify potential pathogens in different Qinhuangdao shorelines.This article combined the PCR gene amplification technology,gel electrophoresis technology and real-time fluorescent quantitative PCR technology to obtain five kinds of pathogenic bacteria to prepare Q-PCR standard curve.Amplification using specificity primer results in linear correlation coefficient of standard curve greater than 0.99 with good reproducibility.Minimum detectable concentration also conforms to the requirements of the environmental samples.We combined chlorophyll measurement technology,automatic nutrient analyzer technology,and flow cytometry instrument to measure the total number of bacteria.Quantitative PCR assays on five representative pathogenic bacteria and correlation analysis were performed with samples collecting in July and October 2015 in Qinhuangdao coastal Luanhe river mouth and Xinkai river mouth.Result shows:N?P significantly affect the estuarine scallop culture zones deputy hemolytic Escherichia coli and vibrio content,with regression equation established to deputy hemolytic Vibrio Y=0.4TN+2PN+19TP+12NO₂^--443?R²=0.491,P<0.001?.Through regulation related to control quantity of ingredients of the nutritional content of the pathogen,realize the prevention and early warning.We combined chlorophyll measurement technology,automatic nutrient analyzer technology,and flow cytometry instrument to measure the total number of bacteria.Quantitative PCR assays on five representative pathogenic bacteria and correlation analysis were performed with samples collecting in January,march,may,July 2016 in qinhuangdao terrigenous outlet of the sea to analysis,the result shows:N?P content in the outlet can promote deputy hemolytic Vibrio breeding;Estuarine environment is more complex,N?P content affects the reproduction of Escherichia coli.The mouth of five kinds pathogenic bacteria to establish regression equation for For E.coli:y=3.3×10⁵+552TP-2169NO₂^-+6.5PN?R²=0.775,P<0.001?;Salmonella:y=1429DP+PN-6816DO-1400?R²=0.794,P<0.001?.To control the content of nitrogen and phosphorus can effectively reduce the abundance of two pathogens.This article combined chlorophyll measurement technology,national standard method for determining nutrient,fluorescence microscope count the total number of bacteria technology,Q-PCR quantitative five kinds of pathogenic bacteria and correlation analysis,regression analysis technology,taking different temperature gradient into account,sampling in January,march,may,July 2016 in qinhuangdao six main baths to analysis,the result shows:pathogenic bacteria have a sudden high value points,five kinds of pathogenic bacteria to establish regression equation of bath for E.coli:y=2.3×10⁵+434TP-104NO₂^-+9PN?R²=0.885,P<0.001?;For Staphylococcus aureus:y=3.7×10⁴+6TP+2500NO₂^-+1.3×10⁵Tem?R²=0.814,P<0.001?.Control the content of nitrogen and phosphorus can effectively reduce the abundance of two pathogens.