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从鱼类和鱼类处理者中分离的多药耐药铜绿假单胞菌的耐药模式、毒力决定因素和生物膜基因。

Resistance patterns, virulence determinants, and biofilm genes of multidrug-resistant Pseudomonas aeruginosa isolated from fish and fish handlers.

机构信息

Department of Zoonoses, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt.

出版信息

Sci Rep. 2024 Oct 14;14(1):24063. doi: 10.1038/s41598-024-73917-4.

Abstract

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic bacterium that is widely distributed in aquatic environments and causes major economic losses in fish and public health hazards.This study aimed to identify the occurrence of P. aeruginosa in samples collected from fish and fish handlers, and to investigate the antimicrobial susceptibility, virulence determinants, and biofilm genes of P. aeruginosa isolates. A total of 276 samples were cross-sectionally collected from Nile tilapia (53), Golden grey mullet (52), Mediterranean horse mackerel (50), Striped red mullet (71), and fish handlers (50) at five different retail fish markets in Damietta Governorate, Egypt. Pseudomonas species (spp.) were biochemically identified in 57.9% of the total examined samples. Peudomonas aeruginosa were the most prevalent species isolated from the fish and human samples via PCR technique. Peudomonas aeruginosa isolates exhibited full resistance (100%) to tobramycin (TOB), gentamicin (CN), and colistin (CL), with a high level of susceptibility (88.5%) to imipenem (IPM) using the disk diffusion method. Most P. aeruginosa isolates (84.6%) exhibited drug resistance, with 61.5% were multidrug resistance (MDR) and 23.1% were extensive drug resistance (XDR). Most isolates had at least four virulence-associated genes (lasB, toxA, exoU, and oprL) and three biofilm genes (psIA, peIA, and lasR) by using uniplex PCR. The lasI, and rhlR Quorum Sensing (QS) genes were identified in 84.6% and 61.5% in the examined P. aeruginosa isolates, respectively. The highest mortality rate in Nile tilapia experimentally infected with P. aeruginosa isolate encoding most of virulent genes. Multivariate analyses revealed high heterogeneity among the examined isolates. This study revealed the emergence of virulent and drug resistant P. aeruginosa isolates in fish, poses high risks to consumers and food. Thus, strict hygienic measures should be considered when catching, handling, and storing fish, in addition to the routine application of antimicrobial susceptibility testing.

摘要

铜绿假单胞菌(P. aeruginosa)是一种机会性病原体,广泛分布于水生环境中,给鱼类养殖造成了重大经济损失,同时也对公共健康构成了威胁。本研究旨在调查埃及达米埃塔省五个不同零售鱼市场采集的鱼类和鱼贩样本中铜绿假单胞菌的发生情况,评估其对抗菌药物的敏感性、毒力决定因素和生物膜基因。共采集了 276 个样本,包括尼罗罗非鱼(53 个)、金梭鱼(52 个)、地中海马鲛鱼(50 个)、条纹红鲷鱼(71 个)和鱼贩(50 个)。通过生化方法鉴定,总样本中有 57.9%为假单胞菌属(spp.)。通过 PCR 技术从鱼类和人类样本中分离出最常见的菌种是铜绿假单胞菌。纸片扩散法检测显示,铜绿假单胞菌对妥布霉素(TOB)、庆大霉素(CN)和黏菌素(CL)完全耐药(100%),对亚胺培南(IPM)高度敏感(88.5%)。大多数铜绿假单胞菌(84.6%)表现出药物耐药性,61.5%为多药耐药(MDR),23.1%为广泛耐药(XDR)。大多数分离株至少有四个与毒力相关的基因(lasB、toxA、exoU 和 oprL)和三个生物膜基因(psIA、peIA 和 lasR),使用单重 PCR 进行检测。lasI 和 rhlR 群体感应(QS)基因在检测到的铜绿假单胞菌分离株中的检出率分别为 84.6%和 61.5%。实验感染携带大多数毒力基因的铜绿假单胞菌分离株的尼罗罗非鱼死亡率最高。多变量分析显示,检测到的分离株之间存在高度异质性。本研究揭示了鱼类中出现了毒力和耐药性的铜绿假单胞菌分离株,对消费者和食品构成了高风险。因此,在捕捞、处理和储存鱼类时应采取严格的卫生措施,此外还应常规进行抗菌药物敏感性试验。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a33/11473961/af4fa06ff771/41598_2024_73917_Fig2_HTML.jpg

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