| This paper studied the isolation and biochemical identification of bacteria and other methods of suspected chicken identification of Klebsiella pneumoniae were cultured, was determined by micro-dilution method for the determination of the antimicrobial agents against clinical isolates of Klebsiella pneumoniae chicken minimum inhibitory concentration （MIC）, followed byβ-lactamase genetypes and subtypes of the analysis to determine the source of Klebsiella pneumoniae producing the chickenβ-lactamase molecular type. To further understand the production CTX-M-14 type ESBL gene and gene environmental structure transmission mechanism, on the production of CTX-M-14 type ESBL genes were plasmid conjugal transfer test; of positive transconjugants were plasmid extraction and digestion, PBR322 Vector and transformed into the fragment is confirmed, the delivery of biological sequencing company, and the Blast sequence alignment. Finally, save the laboratory identification of TEM-57 type containingβ-lactamase gene expression in bacteria inhibiting enzyme hydrolysis protection experiments and tests to detect hydrolysis rates of this enzyme to antibiotics and inhibitors andβ-lactam drugs combined inhibition of the enzyme after the enzyme rate of protection, aimed to provide a theoretical basis for the rational use of antibiotics in veterinary practice.Klebsiella pneumoniae from chicken resistant phenotype, genotype test results showed that only the carbapenems （imipenem）, cefepime and polymyxin K1 of Klebsiella pneumoniae strains with high antibacterial activity; measured K1 strain producing both ESBLs and AmpC enzymes produced by ESBLs as CTX-M-14 type, produced by the AmpC enzyme ACT-like Evolution-type, and also produced TEM-1 and SHV-11 type extended-spectrumβ-lactamase. Will blaCTX-M-14, blaSHV-11 and the Evolution of type blaACT-like gene sequences obtained uploaded to the NCBI sequence numbers were GU211011, GU211012 and GU211014. CTX-M-14 type ESBL-produ- cing plasmid conjugal transfer test results show that CTX-M-14 type ESBL gene-positive test strains to pass through the plasmid conjugal transfer resistance, CTX-M-14 and TEM-1 gene at the same time by plasmid transfer, transconjugants resistant phenotype strains remained the same donor, the donor strains with similar resistance phenotype, only the MIC value than the individual drugs for body of fungi decreased. Cefotaxime-containing screening plates containing CTX-M-14 gene, the positive transformants resistant to cefotaxime, while the rest of the performance was sensitive to most drugs.CTX-M-14 type ESBL-producing genetic environmental results show that: transformant contain 2637 bp gene sequence, through Blast comparison was informed that the gene sequence of the environment into structural elements, including upstream insertion sequence ISEcp1, -35 and -10 region, repeat IRR, 876 bp of the CTX-M-14 gene downstream open reading frame and insertion sequence IS903 and IRL, the sequence has been uploaded to the NCBI sequence number is HQ650134.TEM-57 typeβ-lactamase analysis show that, TEM-57-typeβ-lactamase the Km value of the minimum of cephalexin, the affinity of the strongest, the drug most likely to hydrolysis. Sulbactam and tazobactam inhibition of enzyme amoxicillin protection rate was the highest （87.3% and 92.4%）. Tazobactam to penicillin, amoxicillin, ampicillin and ceftriaxone in the protective effect of enzyme inhibition was stronger than sulbactam, but tazobactam ceftiofur sodium on the inhibition effect of enzyme protection was weaker than sulbactam. |
