Isolation And Identification Of Trueperella Pyogenes And Detection Of Virulence Factors

The Trueperella Pyogenes is a symbiotic bacteria on economic livestock and wild animalssuch as most of the breast in healthy animals, urogenital tract, respiratory tract and gastrointestinaltract, and it is also a conditional pathogens on the mucosa of these animals. It can lead to a varietyof animal purulent infection of many organs and cause metritis, renal abscess, mastitis in dairycows, osteomyelitis, arthritis and other diseases, and serious economic losses to the aquacultureindustry.In this study,3suspected Trueperella Pyogenes isolates have been identified bymorphological characteristics, cultural characteristics, biochemical characteristics.At the same time,16S rRNA gene cloning, sequence analysis has been use in isolates’ identification.Gene sequenceanalysis results by sequence analysis is Trueperella Pyogenes. With the known sequence bysoftware analysis,homology of3isolates and two isolates stored in the laboratory16S rRNAsequence is99.84%.Compared with6isolates from suppurative cows uterine endometritisand5isolates from suppurative pneumonia,values were99.82%and99.96%.3isolates were namedHLJ-1204-F1, HLJ-1204-F3, HLJ-1204-G3.After preliminary identification of virulence factors，5isolates infected mice by intraperitonealinjection with HLJ-1204-F1, HLJ-1204-F3, HLJ-1005. Design primers in order to detect, sequenceand predict the protein structure of virulence genes. Producting whole cell antiserum withHLJ-1204-F1, HLJ-1005,in order to study the role of bacterial protein and whole cell in MDBKcells.The results show that HLJ-1005missing CbpA and contains a NanP, and NanH gene is largermutations at both ends.The CbpA3D structure is similar to Staphylococcus aureus collagen bindingprotein, NanH and NanP comply with the three-dimensional structure of the bacterial sialidasemode.Animal experiments showed HLJ-1005strains have less pathogenic than the other twoisolates.Cell experiments have shown cell adsorption properties of bacterial protein present in thebacterial surface of bacteria,and HLJ-1204-F1have stronger bacterial adsorption.Antiserumsignificantly inhibited the adsorption of the cells.