1株鲤源致病性铜绿假单胞菌的分离鉴定及药敏试验

doi:10.16431/j.cnki.1671-7236.2023.11.037

Abstract

Abstract: 【Objective】 The purpose of this study was to isolate and identify the pathogenic bacteria that caused a large number of acute deaths in carp, determine their virulence genes and drug sensitivity, and provide reference for the subsequent clinical treatment of sick carp.【Method】 The pathogenic bacteria were isolated and cultured from the viscera of diseased carp.Morphological observation, Gram staining, physiological and biochemical identification and 16S rRNA molecular identification were performed.The drug sensitivity and pathogenicity of the pathogen were analyzed by virulence gene detection, K-B drug sensitive disk method and artificial regression infection test.【Result】 The isolated bacteria showed round or oval colonies on LB solid medium.Microscopic examination showed short rod-like Gram-negative bacteria with certain locomotor ability.The results of physiological and biochemical tests showed that the isolated strains showed positive reactions in D-glucose, gelatin, urease, oxidase, arginine, ornithine, lysine and citric acid tests, while the other indexes showed negative reactions, which was consistent with the physiological and biochemical characteristics of Pseudomonas aeruginosa.The phylogenetic tree of 16S rRNA gene sequence showed that the isolated bacteria showed high homology with Pseudomonas aeruginosa NR_114471.1, with 99% similarity.Based on the above test results, the isolated bacteria was identified as Pseudomonas aeruginosa and named HB2210.Virulence gene detection showed that the strain contained several virulence genes, such as flagellin structure gene fla gene, aerolysin aer gene, cytotoxic enterotoxin act gene, outer membrane protein ompA gene and heat stable enterotoxin AST gene. The strain was found to be sensitive to nine antibiotics including ceftazidime, ceftriaxone, cefoperazone, norfloxacin, ofloxacin, ciprofloxacin, polymyxin B, levofloxacin and piperacillin by drug sensitivity testing.And it was found to be acutely virulent to common carp by artificial regression infection tests, with mortality rates of up to 85% in the high concentration group.【Conclusion】 In this experiment, Pseudomonas aeruginosa HB2210 isolated from diseased common carp tissues had strong virulence, and nine antibiotics such as ceftazidime, ceftriaxone, cefoperazone, norfloxacin, ofloxacin, ciprofloxacin, polymyxin B, levofloxacin, and piperacillin, etc.screened out by sensitization test, and could provide the theoretical basis of the precise medication for common carp aquaculture enterprises.

Key words: common carp; Pseudomonas aeruginosa; drug sensitivity test; virulence gene

CLC Number:

S852.61

ZOU Haobo, XUE Shuqun, LYU Xiaonan, GAO Congting, BING Hui, GU Jinzhao, HAN Ying, XIA Banghua. Isolation, Identification and Drug Sensitivity Test of a Pathogenic Strain of Pseudomonas aeruginosa from Common Carp[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4703-4713.

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Isolation, Identification and Drug Sensitivity Test of a Pathogenic Strain of Pseudomonas aeruginosa from Common Carp

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