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Assembly and Characterization of a Pathogen Strain Collection for Produce Safety Applications: Pre-growth Conditions Have a Larger Effect on Peroxyacetic Acid Tolerance Than Strain Diversity

机译:用于生产安全应用的病原体菌株集合的组装和表征:生长前条件对过氧乙酸耐受性的影响大于菌株多样性

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摘要

Effective control of foodborne pathogens on produce requires science-based validation of interventions and control strategies, which typically involves challenge studies with a set of bacterial strains representing the target pathogens or appropriate surrogates. In order to facilitate these types of studies, a produce-relevant strain collection was assembled to represent strains from produce outbreaks or pre-harvest environments, including Listeria monocytogenes (n = 11), Salmonella enterica (n = 23), shiga-toxin producing Escherichia coli (STEC) (n = 13), and possible surrogate organisms (n = 8); all strains were characterized by whole genome sequencing (WGS). Strain diversity was assured by including the 10 most common S. enterica serotypes, L. monocytogenes lineages I–IV, and E. coli O157 as well as selected “non-O157” STEC serotypes. As it has previously been shown that strains and genetic lineages of a pathogen may differ in their ability to survive different stress conditions, a subset of representative strains for each “pathogen group” (e.g., Salmonella, STEC) was selected and assessed for survival of exposure to peroxyacetic acid (PAA) using strains pre-grown under different conditions including (i) low pH, (ii) high salt, (iii) reduced water activity, (iv) different growth phases, (v) minimal medium, and (vi) different temperatures (21°C, 37°C). The results showed that across the three pathogen groups pre-growth conditions had a larger effect on bacterial reduction after PAA exposure as compared to strain diversity. Interestingly, bacteria exposed to salt stress (4.5% NaCl) consistently showed the least reduction after exposure to PAA; however, for STEC, strains pre-grown at 21°C were as tolerant to PAA exposure as strains pre-grown under salt stress. Overall, our data suggests that challenge studies conducted with multi-strain cocktails (pre-grown under a single specific condition) may not necessarily reflect the relevant phenotypic range needed to appropriately assess different intervention strategies.
机译:要有效控制农产品上的食源性病原体,需要对干预和控制策略进行基于科学的验证,这通常包括对挑战性研究进行一系列代表目标病原体或适当替代物的细菌菌株的研究。为了促进这些类型的研究,收集了与农产品相关的菌株集合,以代表来自农产品暴发或收获前环境的菌株,包括单核细胞增生李斯特菌(n = 11),肠沙门氏菌(n = 23),产生志贺毒素的菌株大肠杆菌(STEC)(n = 13),以及可能的替代生物(n = 8);所有菌株均通过全基因组测序(WGS)进行表征。通过包括10种最常见的肠炎链球菌血清型,单核细胞增生李斯特氏菌谱系I–IV和大肠杆菌O157以及选定的“非O157” STEC血清型,可以确保菌株的多样性。如先前已表明,病原体的菌株和遗传谱系在承受不同压力条件下的能力可能会有所不同,因此为每个“病原体组”(例如沙门氏菌,STEC)选择了代表性菌株的子集,并评估了它们的存活率。使用在不同条件下预先生长的菌株暴露于过氧乙酸(PAA)中,这些条件包括(i)低pH,(ii)高盐,(iii)水分活度降低,(iv)不同的生长期,(v)基本培养基和( vi)不同的温度(21°C,37°C)。结果表明,与菌株多样性相比,在三个病原体组中,PAA暴露前的生长条件对细菌减少的影响更大。有趣的是,暴露于盐胁迫(4.5%NaCl)的细菌在暴露于PAA后始终表现出最少的减少;而在暴露于PAA后,细菌的减少最少。但是,对于STEC,在21°C下预生长的菌株与在盐胁迫下预生长的菌株一样耐PAA暴露。总体而言,我们的数据表明,使用多菌株鸡尾酒(在单个特定条件下预先种植)进行的挑战研究不一定能反映适当评估不同干预策略所需的相关表型范围。

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