Extended-spectrum beta-lactamase (ESBL)- and carbapenemase (CP)-producing Klebsiella pneumoniae isolates are a public health concern at clinical level, mainly in Southern European countries. However, there are scarce data on the role of companion animals in the emergence of resistance to clinically relevant antibiotics. Therefore, our study aimed to determine the presence of K. pneumoniae with relevant beta-lactamases in fecal samples from healthy dogs (kennel and house dogs) and sick dogs in seven different hospitals in Portugal. Fecal samples from 125 healthy dogs and 231 sick dogs (one per animal) were collected during April–August 2017. Samples were screened on MacConkey agar supplemented with meropenem, and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was used for K. pneumoniae identification. Genotypic detection of ESBLs or CPs was carried out by PCR/sequencing. Moreover, the presence of other antimicrobial resistance genes and multilocus sequence typing was tested by PCR/sequencing. K. pneumoniae isolates were obtained from 16 tested samples (4.4%), and 3 of them were ertapenem and/or meropenem intermediate/resistant (all of them imipenem susceptible and negative for CP genes). Fifteen K. pneumoniae isolates were ESBL producers, and they carried the following beta-lactamase genes: bla_CTX-M-15+bla_SHV-28 (four isolates, in three cases associated with bla_TEM-1), bla_CTX-M-15+bla_SHV-1 (five isolates, associated with TEM-1 in three cases), and bla_SHV-28+bla_TEM-1 (six isolates). Three ESBL-producing K. pneumoniae isolates of different origins and beta-lactamase genotypes (CTX-M-15+SHV-28, CTX-M-15+SHV-28+TEM-1, or SHV-28+TEM-1) belonged to the lineage ST307, and one isolate was identified as ST15 (CTX-M-15+SHV-1). These findings highlight that dogs are frequent carriers of ESBL-producing K. pneumonia isolates, harboring mostly genes encoding CTX-M-15 or SHV-28, associated in some cases with the high-risk clones ST307 and ST15.

中文翻译：

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从葡萄牙的健康犬和病犬身上分离出的广谱产生β-内酰胺酶的肺炎克雷伯菌。

广谱β-内酰胺酶（ESBL）和碳青霉烯酶（CP）产生的肺炎克雷伯菌分离株在临床上是公共卫生问题，主要在南欧国家中。但是，关于伴侣动物在对临床相关抗生素产生抗药性方面的作用的数据很少。因此，我们的研究旨在确定肺炎克雷伯菌的存在葡萄牙7家不同医院的健康犬（狗和家犬）和患病犬的粪便样本中含有相关的β-内酰胺酶。2017年4月至8月，收集了125只健康犬和231只病犬（每只动物一只）的粪便样本。样本在含有美罗培南的MacConkey琼脂上进行了筛选，并辅以基质辅助激光解吸/电离飞行时间（MALDI-TOF ）用于肺炎克雷伯菌的鉴定。ESBL或CP的基因型检测通过PCR /测序进行。此外，通过PCR /测序测试了其他抗菌素耐药基因和多基因座序列类型的存在。肺炎克雷伯氏菌从16个测试样品（4.4％）获得了分离株，其中3个是厄他培南和/或美洛培南中等/耐药（所有亚胺培南易感且CP基因阴性）。15株肺炎克雷伯菌是ESBL产生者，它们携带以下β-内酰胺酶基因：bla _CTX-M-15 + bla _SHV-28（四种分离物，在三种情况下与bla _TEM-1相关），bla _{CTX-M- 15} + bla _SHV-1（5株，与TEM-1相关，三例），以及bla _SHV-28 + bla _TEM-1（6株）。三种产ESBL的肺炎克雷伯菌不同起源和β-内酰胺酶基因型的分离株（CTX-M-15 + SHV-28，CTX-M-15 + SHV-28 + TEM-1或SHV-28 + TEM-1）属于血统ST307，并且一种分离物被鉴定为ST15（CTX-M-15 + SHV-1）。这些发现表明，狗是产生ESBL的肺炎克雷伯菌分离株的常见携带者，大部分携带编码CTX-M-15或SHV-28的基因，在某些情况下与高风险克隆ST307和ST15相关。