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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应变收集的组装和表征:预生长条件对过氧乙酸耐受的影响比应变多样性

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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:有效控制食品载体的病原体对产量需要基于科学的干预和控制策略的验证,这通常涉及具有代表靶病原体或适当替代剂的一组细菌菌株的攻击性研究。为了促进这些类型的研究,组装了一种相关的应变收集,以代表产生爆发或预收获环境的菌株,包括李斯特菌单核细胞增生(n = 11),沙门氏菌肠(n = 23),滋阴生产大肠杆菌(STEC)(n = 13),以及可能的替代生物(n = 8);所有菌株的特征在于全基因组测序(WGS)。通过包括10个最常见的S.肠溶肠溶液,L.单核细胞增生谱系I-IV和大肠杆菌O157以及选定的“非O157”STEC血清型来确保菌株多样性。如前所述,病原体的菌株和遗传谱系在其存活的能力中可能不同,所以选择并评估每个“病原体组”(例如,沙门氏菌,STEC)的代表性菌株的子集以进行存活通过在不同条件下预生长的菌株暴露于过氧乙酸(PAA),所述菌株包括(i)低pH,(ii)高盐,(iii)降低的水活性,(iv)不同的生长相,(v)最小培养基,和( VI)不同的温度(21°C,37°C)。结果表明,与应变多样性相比,在三种病原体基因群中,在PAA暴露后对细菌还原具有更大的影响。有趣的是,暴露于盐胁迫的细菌(4.5%NaCl)始终显示出Paa暴露后的最少减少;然而,对于STEC,在21℃下预先生长的菌株随着在盐胁迫下预生长的菌株而耐受PAA暴露。总的来说,我们的数据表明,用多重菌株鸡尾酒进行的挑战性研究(在单一特定条件下预先生长)可能不一定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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