Isolation And Identification Of Riemerella Anatipestifer And Amplification Of16s RRNA

Riemerella anatipestifer infection is probably the most economically important disease ofduck farms.The present study according to the ecology principle, bacteria physiological andbiochemical identification method and biological characteristics also focuses on16SrRNA (16S ribosomal RNA) to detect Riemerella anatipestifer between otherorganisms and detecting of Riemerella anatipestifer outer membrane protein A, thetechnique widely used as biomarker and for microbial ecology studies. A total of10Riemerella anatipestifer isolates originally from different Fengcheng duck farms inJiangxi province,The samples were taken from the brain and liver. TSA and TSB wereused to isolation and identification of RA. LB was used as cultural media in the methodof amplifying16S rRNA gene. An E.coli, Salmonella, Haemophilus parasuis, andErysipelothrix rhusiopathiae were used to check the specificity of the RA OmpA, andE.coli, Salmonella, Haemophilus parasuis, and Erysipelothrix rhusiopathiae have beenchosen to check the specificity of the RA by using16S rRNA method. biochemical testsshowed that all the isolates are RA. according to results some isolates showed resistanceto some antibiotics.Two pairs of primers used in this study, pair of primer to detect OmpA. The other pair ofprimers is for16S rRNA. the optimizing temperature, the concentration of theprimer,and DNA sensitivity to get sensitive volume, then got the optimizing parametersfor the PCR reaction for16S rRNA. To amplify16S rRNA TIAN GEL Midipurification kit was used to for the study and sequencing of16S rRNA.The results demonstrate that by amplifying16S rRNA and detecting OmpA its usefultool for the diagnosis of Riemerella anatipestifer infection. And the pathogen couldalso detected in clinical samples from diseased ducks.The aim of this study is isolation and identification of RA in pure culture, clinicalsamples and infected duck brains and livers. For diagnosis of RA infection anddifferentiation between other pathogens which appears together and have the sameclinical signs and lesions (E-coli, salmonella enteric).