Characterisation of AmpC / ESBL genes in some pathogen gram-negatives isolated from clinical cases of livestock and companion animals

IF 0.7 4区农林科学 Q3 VETERINARY SCIENCES

Acta Veterinaria-Beograd Pub Date : 2021-12-01 DOI:10.2478/acve-2021-0036

F. Pehlivanoğlu, E. Sababoglu

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Abstract

Abstract This study was aimed to search and characterize the AmpC and/or ESBL genes of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa isolated from clinical cases of local livestock and companion animals between 2017 and 2019. A total of eight ceftiofur-resistant E. coli (n= 7) and ceftiofur-resistant K. pneumoniae (n= 1) and seven P. aeruginosa were isolated from different cases in local animals. By combination disc method, six E. coli isolates and one K. pneumoniae isolate were found to be ESBL producers. By combination of the disc method and double disc synergy test, no P. aeruginosa isolates were found as ESBL producers. In the agar disc diffusion test (ADDT) performed with cefoxitin and cefoxitin-boronic, only one E. coli was determined as AmpC producer. In ESBL-producing isolates, only the CTX-M class gene was detected by polymerase chain reaction (PCR) and subsequent sequence analysis revealed CTX-M-3 and CTX-M-15 variants. An AmpC positive E. coli isolate was found to carry plasmidic ampC gene in cmy-2 variant from CIT family. It was observed that P. aeruginosa isolates did not carry the plasmidic ampC gene. After the chromosomal ampC gene of one P. aeruginosa was amplified by PCR and sequenced, R79Q and T105A mutations in the chromosomal ampC gene was revealed. This showed that overproduction of the ampC enzyme is involved in the resistance to β-lactams in P. aeruginosa isolates in the study.

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从家畜和伴侣动物的临床病例中分离的一些病原体革兰氏阴性中AmpC/ESBL基因的特征

摘要本研究旨在检索和鉴定2017年至2019年间从当地牲畜和伴侣动物的临床病例中分离出的大肠杆菌、肺炎克雷伯菌和铜绿假单胞菌的AmpC和/或ESBL基因。共从当地动物的不同病例中分离出8株对头孢噻呋耐药的大肠杆菌（n=7）和对头孢噻啶耐药的肺炎克雷伯菌（n=1）以及7株铜绿假单胞菌。通过组合圆盘法，发现6株大肠杆菌和1株肺炎克雷伯菌为ESBL产生菌。采用圆盘法和双圆盘协同试验相结合的方法，未发现铜绿假单胞菌产生ESBL。在用头孢西丁和头孢西丁-硼酸进行的琼脂盘扩散试验（ADDT）中，只有一株大肠杆菌被确定为AmpC产生菌。在产ESBL的分离株中，通过聚合酶链式反应（PCR）仅检测到CTX-M类基因，随后的序列分析显示CTX-M-3和CTX-M-15变体。发现一株AmpC阳性的大肠杆菌分离株在CIT家族的cmy-2变异株中携带质粒AmpC基因。观察到铜绿假单胞菌分离株不携带质粒ampC基因。对一株铜绿假单胞菌的染色体ampC基因进行PCR扩增和测序，发现染色体ampC的R79Q和T105A突变。这表明在本研究中，ampC酶的过量产生与铜绿假单胞菌对β-内酰胺的耐药性有关。

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来源期刊

Acta Veterinaria-Beograd 农林科学-兽医学

CiteScore

1.30

自引率

16.70%

发文量

审稿时长

18-36 weeks

期刊介绍： The Acta Veterinaria is an open access, peer-reviewed scientific journal of the Faculty of Veterinary Medicine, University of Belgrade, Serbia, dedicated to the publication of original research articles, invited review articles, and to limited extent methodology articles and case reports. The journal considers articles on all aspects of veterinary science and medicine, including the diagnosis, prevention and treatment of medical conditions of domestic, companion, farm and wild animals, as well as the biomedical processes that underlie their health.