Isolation And Drug Resistance Evaluation Of Campylobacter And Clostridium Perfringens From Poultry Origin

There are many kinds of zoonotic pathogens that can cause human and many animal diseases,not only is it a serious threat to animal and human food safety,it also has an impact on public health safety.Campylobacter,Clostridium and Vibrio are the main species related to human food-borne diseases,moreover,foodborne diseases will not disappear with economic development or a or science and technology advances.In nature,the host of the Campylobacter is very common,Clostridium perfringens is widely distributed in the natural environment,in the feces,and in the intestines of many people and animals.These pathogens can infect humans and animals in different ways and lead to the occurrence of various diseases,there are not only antimicrobial effects,but also diversity of bacterial resistance.For the moment,reports of human and animal poisoning caused by infection with Campylobacter and Clostridium perfringens in China are not many,the relevant epidemiological data statistics are not detailed as well.In order to compare and analyze the epidemic situation of poultry and wildlife infection in different areas of China,and obtain drug resistance background data,1244 poultry faecal samples,549 healthy birdundefineds faecal samples and 54 samples of heart,liver,spleen,lung,kidney and intestine of 9 dead animals from some provinces in China(Jilin,Liaoning,Jiangxi,Qinghai,Guangdong,Guangxi,Inner Mongolia and other places were collected).Culture and isolation of Campylobacter was conducted by optimizing the culture conditions,Campylobacter was identified by PCR,isolation and identification Clostridium perfringens was carried out by selective culture medium and PCR.Culture and isolation of Campylobacter was conducted by optimizing the culture conditions,Campylobacter was identified by PCR,isolation and identification Clostridium perfringens was carried out by selective culture medium and PCR.Their virulence genes were detected by multiplex PCR,and was determined the biofilm,determination of drug resistance of Clostridium perfringens by drug sensitive tablet and E-test strip,which provided basic data for clinical application.In this study,20 strains of Campylobacter(16 strains of Campylobacter jejuni and 4 strains of Campylobacter coli)from 1244 poultry faecal samples were recovered from six provinces in China(Jilin,Liaoning,Jiangxi,Qinghai,Guangdong,Guangxi),the total separation rate is 1.6%(16/1244).24 strains of C.perfringens were isolated from 123 samples of poultry faeces,the isolation rate was 19.5%(24/123),with 42 strains from 549 migratory birds fecal,the isolation rate was 7.7%(42/549),16 strains from 54 dead migratory animals samples,the isolation rate was 29.63%(16/54),they are from some provinces in China(Jilin,Heilongjiang,Jiangxi and other places),the total isolation rate of C.perfringens was 11.29%(82/726).Clostridium perfringens isolated and identified were tested for biofilm.Among them,30 Clostridium perfringens could form biofilm,the results show that the formation of biofilm is related to the sensitivity of bacteria to antibiotics.Identification of toxicotypes of isolated strains by multiple PCR,according to the results of PCR identification:One toxin E strain was confirmed from night heron of Nanning,Guangxi,and two toxin E strains from shorebirds of Beihai,Guangxi,two toxin E strains from disease tissue of dead birds in Songhua River,Jilin Province,other strains were toxin A strains,however,no ?2 toxinen gene,enterotoxin gene and NetB toxin genes were detected in all strains.The minimal inhibitory concentration(MIC)of antibiotics was determined by using E-test test test strip,criteria for determination of CLSI resistance to anaerobic Clostridium spp.according to the standard of resistance to anaerobic clostridium,sensitivity and drug resistance was analyzed,The percentage of strains reducing sensitivity to Colistin,Clindamycin and Erythromycin were more than 40%,all strains were sensitive to compound sulfamethoxine and Moxifloxacin,the percentage of multi-drug resistant(R?3classes)isolates was 50%(22/44).A total of 6 strains were resistant to more than 5 kinds of antibiotics.Migratory birds carrying C.perfringens were not popular,but strains with high rate of resistance to some antibiotics need to be paid more attention.In this study,poultry samples from different areas were collected,optimization of Campylobacter culture conditions,isolation and identification of Campylobacter were conducted.Their virulence genes were detected,biofilm formation and drug resistance of these C.perfringens strains were evaluated.This study explored the risk of transmissible disease through environment and food chain in migrant birds,to understand the prevalence of drug-resistant bacteria,and laid the foundation for the prevention and control of bacterial drug resistance,provide reference data for food safety monitoring.