Department of Food Science and Human Nutrition, College of Agricultural, Consumer, and Environmental Sciences, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
BMC Microbiol. 2019 Nov 19;19(1):257. doi: 10.1186/s12866-019-1631-3.
The foodborne pathogen Listeria monocytogenes causes the potentially lethal disease listeriosis. Within food-associated environments, L. monocytogenes can persist for long periods and increase the risk of contamination by continued presence in processing facilities or other food-associated environments. Most research on phenotyping of persistent L. monocytogenes' has explored biofilm formation and sanitizer resistance, with less data examining persistent L. monocytogenes' phenotypic responses to extrinsic factors, such as variations in osmotic pressure, pH, and energy source availability. It was hypothesized that isolates of persistent strains are able to grow, and grow faster, under a broader range of intrinsic and extrinsic factors compared to closely related isolates of sporadic strains.
To test this hypothesis, 95 isolates (representing 74 isolates of 20 persistent strains and 21 isolates of sporadic strains) from a series of previous studies in retail delis, were grown at 37 °C, in (i) stress conditions: salt (0, 5, and 10% NaCl), pH (5.2, 7.2, and 9.2), and sanitizer (benzalkonium chloride, 0, 2, and 5 μg/mL) and (ii) energy sources: 25 mM glucose, cellobiose, glycogen, fructose, lactose, and sucrose; the original goal was to follow up with low temperature experiments for treatments where significant differences were observed. Growth rate and the ability to grow of 95 isolates were determined using high-throughput, OD, growth curves. All stress conditions reduced growth rates in isolates compared to control (p < 0.05). In addition, growth varied by the tested energy sources. In chemically defined, minimal media there was a trend toward more isolates showing growth in all replicates using cellobiose (p = 0.052) compared to the control (glucose) and fewer isolates able to grow in glycogen (p = 0.02), lactose (p = 2.2 × 10), and sucrose (p = 2.2 × 10). Still, at least one isolate was able to consistently grow in every replicate for each energy source.
The central hypothesis was rejected, as there was not a significant difference in growth rate or ability to grow for retail deli isolates of persistent strains compared to sporadic strains for any treatments at 37 °C. Therefore, these data suggest that persistence is likely not determined by a phenotype unique to persistent strains grown at 37 °C and exposed to extrinsic stresses or variation in energy sources.
食源性病原体李斯特菌会导致潜在致命的李斯特菌病。在与食物相关的环境中,李斯特菌可以长时间存活,并通过持续存在于加工设施或其他与食物相关的环境中增加污染的风险。大多数关于持久性李斯特菌表型的研究都探索了生物膜形成和消毒剂抗性,而关于持久性李斯特菌对外部因素(如渗透压、pH 值和能源供应的变化)的表型反应的数据较少。据假设,与散发性菌株的密切相关菌株相比,持久性菌株的分离株能够在更广泛的内在和外在因素下生长,并且生长速度更快。
为了验证这一假设,对之前在零售熟食店进行的一系列研究中的 95 株分离株(代表 20 株持久性菌株的 74 株分离株和 21 株散发性菌株)进行了研究,这些分离株在 37°C 下生长,(i)在应激条件下:盐(0、5 和 10%NaCl)、pH(5.2、7.2 和 9.2)和消毒剂(苯扎氯铵,0、2 和 5μg/mL)和(ii)能源源:25mM 葡萄糖、纤维二糖、糖原、果糖、乳糖和蔗糖;最初的目标是对观察到显著差异的治疗方法进行后续低温实验。使用高通量 OD 生长曲线确定 95 株分离株的生长速率和生长能力。所有应激条件都使分离株的生长速度比对照(p<0.05)降低。此外,生长因测试的能源源而异。在化学定义的最小培养基中,与对照(葡萄糖)相比,使用纤维二糖的所有重复试验中,更多的分离株显示出生长趋势(p=0.052),而能够在糖原(p=0.02)、乳糖(p=2.2×10)和蔗糖(p=2.2×10)中生长的分离株减少。尽管如此,至少有一个分离株能够在每个能源源的每个重复中持续生长。
由于在 37°C 下,与散发性菌株相比,零售熟食店的持久性菌株分离株在任何处理下的生长速率或生长能力都没有显著差异,因此该中心假设被拒绝。因此,这些数据表明,持久性可能不是由 37°C 下生长并暴露于外部应激或能源供应变化的持久性菌株特有的表型决定的。