2株鹌鹑源肺炎克雷伯菌的分离鉴定及生物学特性分析

doi:10.16431/j.cnki.1671-7236.2025.05.039

Abstract

Abstract: 【Objective】 The study was conducted to isolate and identify Klebsiella pneumoniae from fecal samples of quail with diarrhea,in order to provide a basis for the prevention and control of Klebsiella pneumoniae disease in quail.【Method】 The purified colonies were screened from 8 fecal samples of quail with diarrhea,and the bacteria were isolated and purified by plate scribing method. 16S rDNA sequence was amplified by PCR and sequenced,and the phylogenetic tree was constructed using Mega 11.0 software.K-B method was used to detect the sensitivity of strains to common antibiotics, the distribution of drug resistance genes and virulence genes in strains was detected,and the biological characteristics was explore.The pathogenicity of the strains was evaluated by animal tests.【Result】 In this study,two strains of Klebsiella pneumoniae were isolated and identified from fecal samples of diarrhea quail,named Ksp-A1 and Ksp-A2,respectively,both belonging to K57 serotype.Phylogenetic tree showed that the two isolates were similar to other known Klebsiella pneumoniae strains,and belonged to the same evolutionary clade as Klebsiella pneumoniae strains DB-4 and WGT-31.The results of drug sensitivity test showed that the isolates were resistant to ampicillin,oxacillin,erythromycin,and so on.Isolates Ksp-A1 and Ksp-A2 carried multiple resistance genes,including bla_SHV,aacC2,oqxAB,tetA,tetM and cat1 genes.The two isolates carried virulence genes such as K88,hlyE, iucD,irp2,ompA and ompC.Biological characteristics analysis showed that in LB broth medium with pH 7.0,isolates Ksp-A1 and Ksp-A2 entered the logarithmic growth phase within 2 h,and reached the peak at 12 and 9 h,respectively.The results of pathogenicity test in mice showed that after the injection of the bacterial solution,the mice showed infection symptoms such as coarse hair,depression,shortness of breath and diarrhea,and the mortality rate of the mice gradually increased with the increase of the bacterial solution concentration,and the mortality rate of the mice reached 100% when the bacterial solution concentration was 1×10⁹ CFU/mL. The median lethal dose (LD₅₀) of isolates Ksp-A1 and Ksp-A2 to mice were 3.3×10⁷ and 2.6×10⁶ CFU,respectively. 【Conclusion】 Two strains of Klebsiella pneumoniae K57 from quail were successfully isolated in the experiment，which showed resistance to β-lactam drugs,had rapid growth rates,and showed high tolerance to acidic and alkaline conditions.The results provided basic data for the prevention and treatment of Klebsiella pneumoniae disease in quail.

Key words: quails; Klebsiella pneumoniae; isolation and identification; drug resistance; virulence genes

CLC Number:

S852.61

LONG Baoqin, WANG Huixiang, YU Linjin, HAERLEHA·Amantai, CHEN Haoran, XU Mengjiao, SHI Longxing, LI Youwen. Isolation,Identification and Biological Characteristics Analysis of Two Strains of Klebsiella pneumoniae from Quail[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(5): 2364-2378.

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Isolation,Identification and Biological Characteristics Analysis of Two Strains of Klebsiella pneumoniae from Quail

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