Dissemination of mcr-1 and β-lactamase genes among Pseudomonas aeruginosa: molecular characterization of MDR strains in broiler chicks and dead-in-shell chicks infections.

IF 4.6 2区 医学 Q1 MICROBIOLOGY
Mona Salem, Gamal Younis, Asmaa Sadat, Nehal Ahmed Talaat Nouh, Dalal Nasser Binjawhar, Mohamed M Abdel-Daim, Mohamed Elbadawy, Amal Awad
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引用次数: 0

Abstract

Objectives: Pseudomonas aeruginosa (P. aeruginosa) is one of the most serious pathogens implicated in antimicrobial resistance, and it has been identified as an ESKAPE along with other extremely significant multidrug resistance pathogens. The present study was carried out to explore prevalence, antibiotic susceptibility phenotypes, virulence-associated genes, integron (int1), colistin (mcr-1), and β-lactamase resistance' genes (ESBls), as well as biofilm profiling of P. aeruginosa isolated from broiler chicks and dead in-shell chicks.

Design: A total of 300 samples from broiler chicks (n = 200) and dead in-shell chicks (n = 100) collected from different farms and hatcheries located at Mansoura, Dakahlia Governorate, Egypt were included in this study. Bacteriological examination was performed by cultivation of the samples on the surface of both Cetrimide and MacConkey's agar. Presumptive colonies were then subjected to biochemical tests and Polymerase Chain Reaction (PCR) targeting 16S rRNA. The recovered isolates were tested for the presence of three selected virulence-associated genes (lasB, toxA, and exoS). Furthermore, the retrieved isolates were subjected to phenotypic antimicrobial susceptibility testing by Kirby-Bauer disc diffusion method as well as phenotypic detection of ESBLs by both Double Disc Synergy Test (DDST) and the Phenotypic Confirmatory Disc Diffusion Test (PCDDT). P. aeruginosa isolates were then tested for the presence of antibiotic resistance genes (ARGs): int1, mcr-1, and ESBL genes (OXA-10, OXA-2, VEB-1, SHV, TEM, and CTX-M). Additionally, biofilm production was examined by the Tube Adherent method (TA) and Microtiter Plate assay (MTP).

Results: Fifty -five isolates were confirmed to be P. aeruginosa, including 35 isolates from broiler chicks and 20 isolates from dead in-shell chicks. The three tested virulence genes (lasB, toxA, and exoS) were detected in all isolates. Antibiogram results showed complete resistance against penicillin, amoxicillin, ceftriaxone, ceftazidime, streptomycin, erythromycin, spectinomycin, and doxycycline, while a higher sensitivity was observed against meropenem, imipenem, colistin sulfate, ciprofloxacin, and gentamicin. ESBL production was confirmed in 12 (21.8%) and 15 (27.3%) isolates by DDST and PCDDT, respectively. Antibiotic resistance genes (ARGs): int1, mcr-1, and ESBL genes (OXA-10, SHV, TEM, and CTX-M), were detected in 87.3%, 18.2%, 16.4%, 69.1%, 72.7%, and 54.5% of the examined isolates respectively, whereas no isolate harbored the OXA-2 or VEB-1 genes. Based on the results of both methods used for detection of biofilm formation, Kappa statistics [kappa 0.324] revealed a poor agreement between both methods.

Conclusions: the emergence of mcr-1 and its coexistence with other resistance genes such as β-lactamase genes, particularly blaOXA-10, for the first time in P. aeruginosa from young broiler chicks and dead in-shell chicks in Egypt pose a risk not only to the poultry industry but also to public health.

铜绿假单胞菌中 mcr-1 和 β-内酰胺酶基因的传播:肉用仔鸡和死雏鸡感染的 MDR 菌株的分子特征。
目的:铜绿假单胞菌(P. aeruginosa)是涉及抗菌药耐药性的最严重病原体之一,它与其他极其重要的多重耐药性病原体一起被确定为 ESKAPE。本研究旨在探讨从肉鸡和壳内死鸡中分离出的铜绿假单胞菌的流行率、抗生素敏感性表型、毒力相关基因、整合素(int1)、可乐定(mcr-1)和β-内酰胺酶耐药基因(ESBls)以及生物膜图谱:本研究从埃及达卡利亚省曼苏拉市的不同农场和孵化场收集了 300 份肉鸡样本(n = 200)和壳内死鸡样本(n = 100)。通过在西曲肽琼脂和麦康凯琼脂表面培养样本,进行细菌学检查。然后对推定菌落进行生化测试和针对 16S rRNA 的聚合酶链式反应(PCR)。对回收的分离物进行检测,以确定是否存在三种选定的毒力相关基因(lasB、toxA 和 exoS)。此外,还采用柯比-鲍尔盘扩散法对回收的分离株进行表型抗菌药敏感性检测,并采用双盘协同试验(DDST)和表型确证盘扩散试验(PCDDT)对 ESBLs 进行表型检测。然后检测铜绿假单胞菌分离物是否存在抗生素耐药基因(ARGs):int1、mcr-1 和 ESBL 基因(OXA-10、OXA-2、VEB-1、SHV、TEM 和 CTX-M)。此外,还采用试管粘附法(TA)和微孔板检测法(MTP)检测了生物膜的产生:结果:55 个分离株被确认为铜绿假单胞菌,其中 35 个来自肉鸡,20 个来自壳内死鸡。在所有分离株中都检测到了三种经测试的毒力基因(lasB、toxA 和 exoS)。抗菌谱结果显示,分离物对青霉素、阿莫西林、头孢曲松、头孢他啶、链霉素、红霉素、广谱霉素和强力霉素完全耐药,而对美罗培南、亚胺培南、硫酸考来烯胺、环丙沙星和庆大霉素的敏感性较高。经 DDST 和 PCDDT 检测,分别有 12 个(21.8%)和 15 个(27.3%)分离菌株证实产生了 ESBL。抗生素耐药基因(ARGs):int1、mcr-1 和 ESBL 基因(OXA-10、SHV、TEM 和 CTX-M)分别在 87.3%、18.2%、16.4%、69.1%、72.7% 和 54.5%的受检分离物中检测到,而没有分离物携带 OXA-2 或 VEB-1 基因。结论:mcr-1 的出现及其与其他耐药基因(如 β-内酰胺酶基因,尤其是 blaOXA-10)的共存,首次出现在埃及幼雏肉鸡和壳内死鸡中的铜绿假单胞菌中,这不仅对家禽业,而且对公众健康都构成了威胁。
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来源期刊
CiteScore
8.60
自引率
0.00%
发文量
49
审稿时长
>12 weeks
期刊介绍: Annals of Clinical Microbiology and Antimicrobials considers good quality, novel and international research of more than regional relevance. Research must include epidemiological and/or clinical information about isolates, and the journal covers the clinical microbiology of bacteria, viruses and fungi, as well as antimicrobial treatment of infectious diseases. Annals of Clinical Microbiology and Antimicrobials is an open access, peer-reviewed journal focusing on information concerning clinical microbiology, infectious diseases and antimicrobials. The management of infectious disease is dependent on correct diagnosis and appropriate antimicrobial treatment, and with this in mind, the journal aims to improve the communication between laboratory and clinical science in the field of clinical microbiology and antimicrobial treatment. Furthermore, the journal has no restrictions on space or access; this ensures that the journal can reach the widest possible audience.
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