The Investigation Of Shellfish Vibrio Disease Epidemiology In Shandong Province And Establishment Of A Rapid Detection Method About Vibrio Parahaemolyticus

Shandong shellfish aquaculture resources, vast area, about 66% of the mariculture area, the annual production that which accounted for about 80% in the provincial total mariculture production is the first in all kinds of aquatic products.But in recent years, along with the rapid development of coastal economy,more and more serious environmental pollution,ecological farming system significantly damaged,fungal diseases are increasingly rampant,some parts of the farming shellfish appeared serious death phenomenon, farmers enthusiasm reduced, production is hampered by a serious ecological farming.Each summer of July- August is the season of farming shellfish microbial disease outbreaks.Therefore,understanding the prevalence of vibrio on shellfish in shangdong province,and establish a relatively rapid detection method is particularly important.Vibrio is one of the main pathogenic bacteria in the breeding of shellfish.Investigating some farming areas in shandong province,the results showed that Yangkou of Weifang,Kenli of Dongying, Zhaoyuan of Yantai,Tianheng of Qingdao and so on appeared the phenomenon of death shellfish.This experiment investigated economic shellfish and wild shellfish of each main breeding in the shandong coastal sea area,by bacteria isolation,cloning and sequence analysis of 16 srRNA,then for identification.Find out main economic shellfish in shandong province in the law of the outbreak of vibrio, build shellfish vibriosis rapid detection method.Results should that altogether collected 68 samples from Yangkou of Weifang;Kenli,Diaokou,Guangli of Dongying and other places.There are 44 samples of the main breeding economic shellfish,24 samples of wild shellfish.151 strains bacteria were isolated from samples,through the analysis of 16 s identification,there are vibrio alginolyticus,vibrio harveyi,vibrio parahaemolyticus,vibrio tubiashii,vibrio vulnificus,aeromonas hydrophila and so on.Separation rate of vibrio is as high as 96%; separation rate of vibrio harveyi in vibrio is as high as 24%, followed by the solution is vibrio alginolyticus 21%.Vibrio harveyi and vibrio parahaemolyticus mainly separat from Dongying, Yantai and other places,vibrio alginolyticus mainly separat from Qingdao, Weihai and other places.The separation rate of vibrio harveyi is higher on shellfish in shandong province, the second is vibrio alginolyticus.Vibrio harveyi mainly separat on scallops,ruditapes philippinarum,scapharca subcrenata.Vibrio parahaemolyticus mainly separat on ruditapes philippinarum,scapharca subcrenata.vibrio alginolyticus mainly separat on ostrea gigas thunberg,scapharca broughtonii.Vibrio harveyi in each type is relatively common.Experimental results show that vibrio is one of the main pathogenic bacteria on economic shellfish in shandong.Particularly,it damaged serious on scallops,ostrea gigas thunberg,scapharca subcrenata,scapharca broughtonii,ruditapes philippinarum and so on.Vibriosis occurs mainly in high temperature season(July, August),this consistent with bacteria optimum growth temperature.Vibrio parahemolyticus in food detection method specified in the national food safety standards, the visible state of its attention.Purpose of this experiment is to establish a rapid detection method of vibrio parahemolyticus in shellfish, not only to ensure the food safety, also facilitate shellfish vibrio parahemolyticus disease detection.Test three pairs of specific primers was designed on not heat-resistant hemolytic toxin genes of vibrio parahemolyticus, according to the principle of ring mediated isothermal amplification techniques(LAMP) for amplification.Select 20 strains of vibrio parahemolyticus and 20 strains of other vibrio for amplification, results show that the LAMP test method of strong specificity, also shows high sensitivity comparison with ordinary PCR detection methods,88fg/LAMP.Therefore, research shows that the LAMP test method is of high sensitivity, specificity, and short time(about 1 h), simple operation, to be an effective methoddetection of shellfish vibrio parahemolyticus disease detection.