Laboratoire de Biotechnologie des Molécules Bioactives et de la Physiopathologie Cellulaire (LBMBPC), Faculté des Sciences de la Nature et de la Vie, Université Batna 2, Batna 05000, Algeria.
Département de Biologie, Université Abou Bekr Belkaid-, Tlemcen 13000, Algeria.
Sci Total Environ. 2022 Mar 25;814:152861. doi: 10.1016/j.scitotenv.2021.152861. Epub 2022 Jan 5.
β-lactams and colistin resistance in Enterobacterales is a global public health issue. In this study we aimed to investigate the occurrence and genetic determinants of Extended-Spectrum β-lactamases, carbapenemases and mcr-encoding-genes in Enterobacterales isolates recovered from the migratory bird species Ciconia ciconia in an Algerian city. A total of 62 faecal samples from white storks were collected. Samples were then subjected to selective isolation of β-lactams and colistin-resistant-Enterobacterales. The representative colonies were identified using Matrix-Assisted Laser Desorption-Ionisation Time-of-Flight Mass Spectrometry. Susceptibility testing was performed using the disk-diffusion method. ESBL, carbapenemases, and colistin resistance determinants were searched for by PCR and sequencing. The clonality relationships of the obtained isolates were investigated by multilocus sequence typing assays. Mating experiments were carried out to evaluate the transferability of the carbapenemase and mcr-genes. Forty-two isolates were identified as follows: Escherichia coli (n = 33), Klebsiella pneumoniae (n = 4), Proteus mirabilis (n = 4) and Citrobacter freundii (n = 1). Molecular analysis showed that twelve isolates carried the bla genes alone, fifteen E. coli isolates were positive for the bla gene, six isolates were NDM-5-carriers (two P. mirabilis, two K. pneumoniae and two E. coli) and eight E. coli strains were positive for the mcr-1 gene. MLST results showed a high clonal diversity, where NDM-5-producing strains were assigned to two sequence types (ST167 for E. coli and ST198 for K. pneumoniae), whereas the mcr-1 positive E. coli isolates belonged to ST58, ST224, ST453, ST1286, ST2973, ST5542, ST9815 and the international high-risk resistant lineage ST101. To the best of our knowledge, this is the first report of bla gene in white storks and also the first describing the mcr-1 gene in white storks in Algeria. This study underlines the important role of migratory white storks as carriers of high-level drug-resistant bacteria, allowing their possible implication as indicators and sentinels for antimicrobial resistance surveillance.
β-内酰胺类和黏菌素耐药性在肠杆菌科是一个全球性的公共卫生问题。在这项研究中,我们旨在调查从阿尔及利亚城市的迁徙鸟类物种白鹳中分离的肠杆菌科分离株中出现的和遗传决定因素,包括超广谱β-内酰胺酶、碳青霉烯酶和 mcr 编码基因。从白头鹤中采集了 62 份粪便样本。然后对选择性分离β-内酰胺类和黏菌素耐药性肠杆菌科进行了处理。使用基质辅助激光解吸电离飞行时间质谱法对代表性菌落进行了鉴定。使用纸片扩散法进行了药敏试验。通过 PCR 和测序来搜索 ESBL、碳青霉烯酶和黏菌素耐药决定因素。通过多位点序列分型试验来研究获得的分离株的克隆关系。进行交配实验以评估碳青霉烯酶和 mcr 基因的可转移性。鉴定出 42 株分离株,分别为:大肠埃希菌(n=33)、肺炎克雷伯菌(n=4)、奇异变形杆菌(n=4)和弗氏柠檬酸杆菌(n=1)。分子分析表明,有 12 株菌单独携带 bla 基因,15 株大肠埃希菌携带 bla 基因,6 株菌为 NDM-5 携带菌(2 株奇异变形杆菌、2 株肺炎克雷伯菌和 2 株大肠埃希菌),8 株大肠埃希菌为 mcr-1 阳性。MLST 结果显示高度克隆多样性,其中产 NDM-5 的菌株被分为两种序列类型(大肠埃希菌为 ST167,肺炎克雷伯菌为 ST198),而 mcr-1 阳性的大肠埃希菌分离株属于 ST58、ST224、ST453、ST1286、ST2973、ST5542、ST9815 和国际高风险耐药谱系 ST101。据我们所知,这是首次在白鹳中发现 bla 基因的报道,也是首次在阿尔及利亚的白鹳中描述 mcr-1 基因。本研究强调了迁徙白鹳作为高水平耐药菌携带者的重要作用,使其可能成为抗菌药物耐药性监测的指标和哨兵。