The Drug Resistant Analysis Of Pathogenic Aeromonas Hydrophila Mediated By Class ? Integron And Preliminary Study Of R Plasmid

Aeromonas hydrophila(AH)is a pathogenic bacterium which affecting humans,animals and fish.At present,the drug resistance of AH has showed a rapid growth trend,new researches find that,integron plays an important role in disseminating multi-drug resistant gene between bacteria,which cause great difficulties in the prevention of AH disease.At the same time,in order to solute the spread of drug resistance,Chinese herbal medicine and natural plants become the focus of attention.Based on these study,this test is aimed at establishing a rapid detection of AH and analyzing its drug resistance,the content as follows:1.A multiplex PCR for detection of aerolysin and class I integron of AHThis study established a multiplex PCR for detection of 16SrRNA,aerolysin and class I integron.Through detecting the suspected pathogen from soft-shelled turtle,monopterus,giant salamander,crucian and rana spinosa david,27 strains were identified as AH,among these strains,the ratio of aerolysin positive strains was 70.37%(19/27),and class I integron positive strains was 44.44%(12/27).Through antibiotic sensitive test,we found that all of the strains presented a seriou,multiple drug resistance.In addition to the eight strains' drug resistant rates were 100%,the rest strains' drug resistant rates were same,the class I integron positive strains'drug resistant rates were higher than the negative ones,which explained that the existence of integron may result to the increase of drug resistance.2.The location of class I integronWe adopted spot hybridization and PCR amplification to position class I integron of the positive strains.The results of the two methods were exactly same,10 strains,class I integron were located in chromosome and plasmid,and the other two were only located in chromosome.3.The location of drug resistant geneUsing garlic and aloe to eliminate the resistant plasmid of AH1 which had 3 kinds of plasmids,the rate reached to 90%.Through plasmid transformation and elimination experiments,the results of antibiotic sensitive test showed that,the gene resistanted ceftriaxone,amikacin,cefoperazone,furazolidone,streptomycin,norfloxacin,enrofloxacin and rifampicin were located in the smaller plasmid;the gene resistanted gentamicin,SMZ-TMP,doxycycline,tobramycin,ampicillin,amoxicillin,SIZ and tetracycline were located in the bigger plasmid;the gene resistanted cefazolin,penicillin and erythromycin were located in chromosome.