鸭源奇异变形杆菌和摩氏摩根菌的分离鉴定、NDM耐药基因检测及中药体外抑菌试验

doi:10.12160/j.issn.1672-5190.2023.05.017

摘要/Abstract

摘要：

[目的]确定引起广东省某鸭场鸭群发病的细菌性病原，调查分离菌株对抗菌药物的敏感性及NDM耐药基因携带情况，并筛选出针对分离菌株具有较好体外抑制作用的单味中药。[方法]采集发病鸭只的肝脏、脾脏等病料样本进行细菌分离培养，采用16S rDNA序列PCR扩增及测序比对方法确定分离菌株的种属，利用纸片扩散法（K-B法）测定分离菌株对13种抗菌药物的敏感性，应用PCR法检测10种NDM耐药基因在分离菌株中的分布情况，采用肉汤二倍稀释法测定14种单味中药对分离菌株的最小抑菌浓度（MIC）。[结果]经革兰染色镜检、生化鉴定及16S rDNA序列分析，从发病鸭病料中分离、鉴定出1株奇异变形杆菌（SK1株）和1株摩氏摩根菌（SK2株）。SK1株和SK2株均对新霉素、头孢他啶、头孢噻肟、头孢哌酮敏感，对多西环素、复方新诺明、恩诺沙星、氟苯尼考、多黏菌素B耐药。SK1株10种NDM耐药基因均未检出，SK2株携带bla_NDM_-1基因，其他9种NDM耐药基因未检出。博落回对SK1株的抑菌效果最好，MIC为3.906 mg/mL；连翘对SK2株的抑菌效果最好，MIC为3.906 mg/mL。[结论]引起该鸭场鸭群发病的主要病原菌为奇异变形杆菌和摩氏摩根菌，可使用新霉素、头孢菌素类抗菌药物及中药博落回、连翘对患病鸭只进行治疗。

关键词: 鸭, 分离鉴定, NDM耐药基因, 奇异变形杆菌, 摩氏摩根菌, 中药, 体外抑菌作用

Abstract:

[Objective] The aims of the present study were to identify the bacterial pathogens causing the disease in a duck farm in Guangdong Province, to characterize the antimicrobial susceptibility and the presence of NDM resistance genes of the isolated strains, and to screen the single traditional Chinese medicine with in vitro bacteriostatic effects against the isolated strains. [Method] The clinical samples of liver and spleen from the diseased ducks were collected for bacterial isolation and cultivation. The PCR amplification, sequencing and similarity comparison of 16S rDNA gene were used to identify the species of the isolated strains. The susceptibility test of the isolated strains to 13 antibacterial agents was carried out with K-B disk diffusion method. The PCR assay was used to detect the distribution of 10 NDM resistance genes in the isolated strains. The broth double dilution method was employed to determine the minimum inhibitory concentration （MIC） of 14 single Chinese traditional medicines against the isolated strains. [Result] Two bacterial strains were obtained from the clinical samples of the diseased ducks. Through Gram staining and microscopy examination, biochemical identification, as well as 16S rDNA sequence analysis, they were identified as Proteus mirabilis （SK1 strain） and Morganella morganii （SK2 strain）, respectively. Both SK1 and SK2 strains were susceptible to neomycin, ceftazidime, cefotaxime, and cefoperazone, and resistant to doxycycline, co-trimoxazole, enrofloxacin, flufenicol, and polymyxin B. None of the 10 NDM resistance genes were detected in SK1 strain. SK2 strain carried the bla_NDM_-1 gene, while the other 9 NDM resistance genes were not detected. Macleaya cordata （Willd.） R. Br. had the best bacteriostatic effect against SK1 strain, with a MIC of 3.906 mg/mL. Forsythia suspensa （Thunb）. Vahl had the best bacteriostatic effect against SK2 strain, with a MIC of 3.906 mg/mL. [Conclusion] The main pathogens causing the disease in the duck farm were Proteus mirabilis and Morganella morganii. Neomycin, cephalosporin antibiotics, as well as traditional Chinese medicine Macleaya cordata （Willd.） R. Br. and Forsythia suspensa （Thunb）. Vahl were recommended to therapy the diseased ducks.

Key words: duck, isolation and identification, NDM resistance gene, Proteus mirabilis, Morganella morganii, traditional Chinese medicine, in vitro bacteriostatic effect

中图分类号:

石玉节,柴贝贝,张彦昕. 鸭源奇异变形杆菌和摩氏摩根菌的分离鉴定、NDM耐药基因检测及中药体外抑菌试验[J]. 畜牧与饲料科学, 2023, 44(5): 121-128.

SHI Yujie,CHAI Beibei,ZHANG Yanxin. Isolation，Identification and Detection of NDM Resistance Genes of Proteus mirabilis and Morganella morganii from Ducks, and in vitro Antibacterial Test of Traditional Chinese Medicine[J]. Animal Husbandry and Feed Science, 2023, 44(5): 121-128.

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