Maamar Elaa, Hammami Samia, Alonso Carla Andrea, Dakhli Nouha, Abbassi Mohamed Salah, Ferjani Sana, Hamzaoui Zaineb, Saidani Mabrouka, Torres Carmen, Boutiba-Ben Boubaker Ilhem
University of Tunis El Manar, Faculty of Medicine of Tunis-Research Laboratory «Antimicrobial resistance», Tunis, Tunisia.
University of Tunis El Manar, Faculty of Medicine of Tunis-Research Laboratory «Antimicrobial resistance», Tunis, Tunisia; University of Gafsa, Faculty of Sciences of Gafsa, Tunisia.
Int J Food Microbiol. 2016 Aug 16;231:69-75. doi: 10.1016/j.ijfoodmicro.2016.05.001. Epub 2016 May 3.
This study was conducted to detect extended spectrum beta-lactamases (ESBLs) and plasmidic AmpC beta-lactamase (pAmpC-BL)-producing Escherichia coli isolates in industrial poultry samples were collected from healthy chickens of the three farms. Samples were inoculated onto desoxycholate-lactose-agar plates supplemented with cefotaxime (2mg/L). E. coli was identified by biochemical and molecular methods and antibiotic susceptibility testing by the disk diffusion method. Genes encoding ESBLs and pAmpC-BL were detected by PCR and sequencing. Phylogenetic groups were determined by triplex PCR. The molecular typing of strains was done by pulsed field gel electrophoresis (PFGE) and Multilocus Sequence Typing (MLST) in those isolates showing different PFGE patterns. Cefotaxime-resistant E. coli isolates were recovered in 48 of 137 fecal samples (35%), and one isolate/sample was further studied. The following beta-lactamase genes were detected: blaCTX-M-1 (29 isolates, isolated in all three farms), blaCTX-M-15 (5 isolates, confined in farm II), blaCTX-M-14 and blaCMY-2 (one isolate and 13 isolates, respectively, in farm III). The 48 cefotaxime-resistant isolates were distributed into phylogroups: B1 (n=21), A (n=15) and D (n=12). PFGE analysis revealed 19 unrelated patterns: 15 different profiles among ESBL-positive strains and 4 among the CMY-2-positive isolates. The following sequence types-associated phylogroups were detected: a) CTX-M-1-positive strains: lineages ST542-B1, ST212-B1, ST58-B1, ST155-B1 and ST349-D; b) CTX-M-15-positive strain: lineage ST405-D; c) CTX-M-14-positive strain: lineage ST1056-B1; d) CMY-2-positive strains: lineages ST117-D, ST2197-A, and ST155-B1. Healthy chickens constitute an important reservoir of ESBL- and pAmpC-BL-producing E. coli isolates that potentially could be transmitted to humans via the food chain or by direct contact.
本研究旨在检测从三个养殖场的健康鸡群采集的工业家禽样本中产生超广谱β-内酰胺酶(ESBLs)和质粒介导的AmpCβ-内酰胺酶(pAmpC-BL)的大肠杆菌分离株。将样本接种到补充有头孢噻肟(2mg/L)的脱氧胆酸盐-乳糖琼脂平板上。通过生化和分子方法鉴定大肠杆菌,并采用纸片扩散法进行抗生素敏感性试验。通过PCR和测序检测编码ESBLs和pAmpC-BL的基因。通过三重PCR确定系统发育群。对那些显示不同PFGE图谱的分离株,通过脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)进行菌株的分子分型。在137份粪便样本中的48份(35%)中分离出对头孢噻肟耐药的大肠杆菌,每份样本分离出一株并进一步研究。检测到以下β-内酰胺酶基因:blaCTX-M-1(29株,在所有三个养殖场均有分离),blaCTX-M-15(5株,仅在养殖场II分离到),blaCTX-M-14和blaCMY-2(分别在养殖场III分离到1株和13株)。48株对头孢噻肟耐药的分离株分布在以下系统发育群中:B1(n = 21)、A(n = 15)和D(n = 12)。PFGE分析显示有19种不相关的图谱:ESBL阳性菌株中有15种不同的图谱,CMY-2阳性分离株中有4种。检测到以下与序列类型相关的系统发育群:a)CTX-M-1阳性菌株:谱系ST542-B1、ST212-B1、ST58-B1、ST155-B1和ST349-D;b)CTX-M-15阳性菌株:谱系ST405-D;c)CTX-M-14阳性菌株:谱系ST1056-B1;d)CMY-2阳性菌株:谱系ST117-D、ST2197-A和ST155-B1。健康鸡是产生ESBL和pAmpC-BL的大肠杆菌分离株的重要储存宿主,这些分离株有可能通过食物链或直接接触传播给人类。