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模仿家禽屠宰场条件对弯曲杆菌属分离株生物膜形成的评估。

Assessment of biofilm formation by Campylobacter spp. isolates mimicking poultry slaughterhouse conditions.

作者信息

Araújo P M, Batista E, Fernandes M H, Fernandes M J, Gama L T, Fraqueza M J

机构信息

CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon 1300-477, Portugal.

CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon 1300-477, Portugal.

出版信息

Poult Sci. 2022 Feb;101(2):101586. doi: 10.1016/j.psj.2021.101586. Epub 2021 Nov 8.

Abstract

This research aimed to assess the biofilm formation ability of Campylobacter strains under temperature and oxygen stress conditions, similar to those found in the industrial environment, to explain the persistence of this pathogen on the poultry slaughter line. A collection of C. jejuni and C. coli isolates (n = 143) obtained from poultry samples (cecal content and neck skin), collected at slaughterhouse level, from diverse flocks, on different working days, was genotyped by flaA-restriction fragment length polymorphism (RFLP) typing method. A clustering analysis resulted in the assignment of 10 main clusters, from which 15 strains with different flaA-RFLP genotypes were selected for the assessment of biofilm formation ability and antimicrobial susceptibility. Biofilm assays, performed by crystal violet staining method, were conducted with the goal of mimicking some conditions present at the slaughterhouse environment, based on temperature, atmosphere, and contamination levels. Results indicated that many C. jejuni strains with similar flaA-RFLP profiles were present at the slaughterhouse on different processing days. All the strains tested (n = 15) were multidrug-resistant except for one. Biofilm formation ability was strain-dependent, and it appeared to have been affected by inoculum concentration, temperature, and tolerance to oxygen levels. At 10°C, adherence levels were significantly lower than at 42°C. Under microaerobic and aerobic atmospheres, at 42°C, 3 strains (C. jejuni 46E, C. jejuni 61C, and C. coli 65B) stood out, exhibiting significant levels of biofilm formation. C. jejuni strains 46E and 61C were inserted in clusters with evidence of persistence at the slaughterhouse for a long period of time. This study demonstrated that Campylobacter strains from broilers are capable of forming biofilms under conditions resembling the slaughterhouse environment. These results should be seen as a cue to improve the programs of hygiene implemented, particularly in those zones that can promote biofilm formation.

摘要

本研究旨在评估弯曲杆菌菌株在温度和氧气胁迫条件下形成生物膜的能力,这些条件类似于工业环境中发现的条件,以解释这种病原体在家禽屠宰线上的持久性。通过flaA限制性片段长度多态性(RFLP)分型方法,对从屠宰场水平收集的、来自不同鸡群、在不同工作日采集的家禽样本(盲肠内容物和颈部皮肤)中获得的空肠弯曲菌和结肠弯曲菌分离株(n = 143)进行基因分型。聚类分析产生了10个主要聚类,从中选择了15个具有不同flaA-RFLP基因型的菌株,用于评估生物膜形成能力和抗菌药敏性。基于温度、气氛和污染水平,采用结晶紫染色法进行生物膜检测,目的是模拟屠宰场环境中存在的一些条件。结果表明,在不同的加工日,屠宰场存在许多具有相似flaA-RFLP谱的空肠弯曲菌菌株。除一株外,所有测试菌株(n = 15)均对多种药物耐药。生物膜形成能力因菌株而异,似乎受到接种物浓度、温度和对氧气水平的耐受性的影响。在10°C时,黏附水平显著低于42°C时。在42°C的微需氧和有氧气氛下,3株菌株(空肠弯曲菌46E、空肠弯曲菌61C和结肠弯曲菌65B)表现突出,表现出显著的生物膜形成水平。空肠弯曲菌菌株46E和61C被归入在屠宰场长期存在的聚类中。本研究表明,来自肉鸡的弯曲杆菌菌株能够在类似于屠宰场环境的条件下形成生物膜。这些结果应被视为改进实施的卫生计划的一个提示,特别是在那些可能促进生物膜形成的区域。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2379/8664863/e174b1f85571/gr1.jpg

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