Isolation and Identification of Klebsiella pneumoniae and Pasteurella multocida and Detection of Drug Resistance in an Intensive Sheep Farm in Shandong Province
In order to identify the pathogens causing cough,polypnea and diarrhea in sheep in an intensive farm in Yanggu County,Shandong Province,nasal pharyngeal swabs and tissue samples of lungs,spleens,livers and tracheas collected from dead sheep to isolate and identify bacteria using morphology observation,biochemical characterization,16S rRNA PCR and genetic evolutionary analysis,followed by test of drug resistance and pathogenicity.The results showed that two kinds of bacteria were isolated and named as LCh/2023-1 and XT/2023-2,respectively,with bacterial culture and biochemical characteristics in compliance with Pasteurella multocida and Klebsiella pneumoniae,respectively;phylogenetic tree revealed that LCh/2023-1 was the highest identical with Pasteurella multocida strain J-4103,and XT/2023-2 was the highest identical with Klebsiella pneumoniae strain HE1;drug resistance analysis results showed that LCh/2023-1 was resistant to cotrimoxazole,ampicillin,florfenicol,kanamycin,neomycin,tetracycline,enrofloxacin,erythromycin and azithromycin,and XT/2023-2 was resistant to cotrimoxazole,carbenicillin,florfenicol,kanamycin,neomycin,tetracycline,doxycycline,enrofloxacin and azithromycin;mouse pathogenicity test results showed that all mice died after the challenge.In conclusion,the disease was caused by mixed infection of Klebsiella pneumoniae and Pasteurella multocida,and the isolates were more virulent with strong drug resistance.
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