中华草龟恶臭假单胞菌的分离鉴定及药物敏感性分析

doi:10.16431/j.cnki.1671-7236.2019.10.034

Abstract

Abstract: In order to find out the pathogen of Chinemys reevesii which was sent for examination from a turtle farm in Haikou city,a dominant growth bacteria (named as J1) was isolated from diseased Chinemys reevesii.Colony culture,Gram staining observation,physiological and biochemical identification and 16S rRNA sequence alignment analysis were used to identify the isolotes,and the pathogenicity and drug sensitivity were verified by virus attack and drug resistance test.The results showed that the strain formed round,smooth,wet,sticky,grey-yellow and opaque colonies on the agar medium.The results of Gram staining microscopy showed that the isolate was short rod-shaped Gram-negative.Physiological and biochemical test results showed that the isolates were sporadic,fructose,citrate,nitrate,oxidase,contact enzyme and arginine hydrolase were positive,while glucose gas production,glucose acid production,ornithine decarboxylase,lactose,maltose,V-P test were negative.16S rRNA gene sequence analysis showed that the isolate had the highest homology with Pseudomonas spp.registered on GenBank.The phylogenetic tree showed that the isolate was clustered with Pseudomonas putida.Artificial challenge test confirmed that the isolated bacteria could cause the disease and death of Chinemys reevesii,and the clinical symptoms and pathological changes were similar to those of naturally occurring turtles.Drug resistance analysis showed that the isolates were sensitive to tobramycin,cefradine and gentamicin,and resistant to clindamycin,tetracycline,ampicillin and cefaclor.The results could provide strong technical support and practical guidance for the diagnosis,prevention and treatment of turtle diseases.

Key words: Chinemys reevesii; Pseudomonas putida; isolation and identification; 16S rRNA; pathogenicity; drug sensitivity

CLC Number:

S947

FANG Zhenhua, DAI Xiaomei, DING Li, HONG Meiling. Isolation,Identification and Drug Susceptibility Analysis of Pseudomonas putida from Chinemys reevesii[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(10): 3115-3123.

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Isolation,Identification and Drug Susceptibility Analysis of Pseudomonas putida from Chinemys reevesii

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