College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi, Xinjiang, China.
Urumqi Animal Disease Control and Diagnosis Center, Urumqi, Xinjiang, China.
Vet Med Sci. 2025 Jan;11(1):e70120. doi: 10.1002/vms3.70120.
Klebsiella pneumoniae is an opportunistic pathogen that causes severe infections in humans and animals. Nonetheless, little is known about the molecular epidemiology of mastitis-associated K. pneumoniae in dairy cows.
This cross-sectional study investigated the epidemiology and antimicrobial resistance (AMR) of K. pneumoniae in 700 milk samples collected from cows with and without mastitis in seven dairy farms in Xinjiang, China.
K. pneumoniae was identified by PCR amplification of the khe gene and the automated VITEK 2 Compact System. Resistance against 18 antimicrobial agents was analysed by broth microdilution. Forty-four new strains were sequenced by whole-genome sequencing (WGS). WGS data were searched for the presence of AMR and virulence genes. Genotypic characterization was performed by multilocus sequence typing and the analysis of wzi allele types and K and O antigens.
K. pneumoniae isolates were found in 131 samples (18.7%). The prevalence of K. pneumoniae in cows with clinical and subclinical mastitis was higher than that in healthy cows (27.1%, 23.2% and 7.3%, respectively). WGS identified 27 wzi allele types, 16 K antigen serotypes, 6 O antigen serotypes and 25 sequence types. Phylogenetic analysis showed high genomic diversity in K. pneumoniae. The rate of resistance to tetracycline and cefazolin was 39.7% and 31.3%, and the multidrug resistance rate was 26.7%. Thirty-nine AMR genes conferring resistance to nine antibiotic classes and 57 virulence genes were identified in new isolates. AMR and virulence genes were more prevalent in known human isolates than in new isolates.
These results improve our understanding of the epidemiology and resistance status of mastitis-associated K. pneumoniae strains. The emergence and spread of multidrug-resistant K. pneumoniae strains threaten food safety and public health.
肺炎克雷伯菌是一种机会致病菌,可导致人类和动物的严重感染。然而,关于奶牛乳腺炎相关肺炎克雷伯菌的分子流行病学知之甚少。
本横断面研究调查了中国新疆 7 个奶牛场 700 份乳腺炎和非乳腺炎奶牛牛奶样本中肺炎克雷伯菌的流行病学和抗菌药物耐药性(AMR)。
通过 khe 基因和自动 VITEK 2 Compact 系统的 PCR 扩增鉴定肺炎克雷伯菌。通过肉汤微量稀释法分析对 18 种抗菌药物的耐药性。对 44 株新菌株进行全基因组测序(WGS)。通过 AMR 和毒力基因的存在情况搜索 WGS 数据。通过多位点序列分型和 wzi 等位基因类型和 K 和 O 抗原分析进行基因型特征分析。
在 131 份样本(18.7%)中发现了肺炎克雷伯菌分离株。乳腺炎和亚临床乳腺炎奶牛中肺炎克雷伯菌的检出率高于健康奶牛(分别为 27.1%、23.2%和 7.3%)。WGS 确定了 27 个 wzi 等位基因类型、16 个 K 抗原血清型、6 个 O 抗原血清型和 25 个序列型。系统发育分析显示肺炎克雷伯菌具有高度的基因组多样性。对四环素和头孢唑林的耐药率分别为 39.7%和 31.3%,多药耐药率为 26.7%。在新分离株中发现了 39 种耐药基因,可赋予对 9 类抗生素的耐药性,以及 57 种毒力基因。与新分离株相比,已知的人类分离株中 AMR 和毒力基因更为普遍。
这些结果提高了我们对乳腺炎相关肺炎克雷伯菌菌株的流行病学和耐药现状的认识。多药耐药肺炎克雷伯菌菌株的出现和传播威胁着食品安全和公共健康。