Department of Veterinary Disease Biology, Centre for Advanced Food Studies, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark.
Foodborne Pathog Dis. 2010 Sep;7(9):1013-8. doi: 10.1089/fpd.2009.0470.
The objective of this study was to investigate how various growth conditions influence the virulence of Listeria monocytogenes monitored by its ability to invade the epithelial cell lines Caco-2 and INT-407. The growth conditions examined were modified atmosphere-packaged deli meat and brain heart infusion broth (BHI) with and without salt. Five strains of L. monocytogenes were selected to investigate their invasiveness and all strains invaded Caco-2 cells at higher levels than INT-407 cells. Further, the clinical strains (3443 and 3734) were more invasive (p < 0.05) than the strains isolated from meat and food-processing environments (3008, 3126, and 4140) after grown in BHI at 30 degrees C. This attenuation could not be ascribed to a defective Internalin A as all strains encoded an intact inlA gene. To determine the influence of food products on virulence, the ability of L. monocytogenes to invade Caco-2 cells was compared after growth on a fermented sausage and on cured cooked ham to that of bacteria grown in BHI broth supplemented with salt. Samples were stored under chilling conditions for up to 4 weeks. The results showed no difference (p > 0.05) in invasiveness after 7 days at 10 degrees C in BHI broth or on sausage, whereas a slight increase (p < 0.05) was observed after incubation on ham for 2 and 4 weeks compared to that in BHI broth. Most importantly, our results show that L. monocytogenes efficiently invade Caco-2 cells even after 4 weeks of storage at chilled temperature. This is highly relevant for safety assessment of this organism in food as these conditions reflect storage of ready-to-eat food products in domestic refrigerators.
本研究旨在探讨不同生长条件如何通过李斯特菌侵袭上皮细胞系 Caco-2 和 INT-407 的能力来影响其毒力。检查的生长条件为改良气氛包装熟食肉和脑心浸液肉汤(BHI),有盐和无盐。选择了五株李斯特菌来研究其侵袭性,所有菌株对 Caco-2 细胞的侵袭水平均高于 INT-407 细胞。此外,临床分离株(3443 和 3734)在 30°C 下于 BHI 中生长时比从肉类和食品加工环境中分离的菌株(3008、3126 和 4140)更具侵袭性(p < 0.05)。这种衰减不能归因于 Internalin A 的缺陷,因为所有菌株都编码了完整的 inlA 基因。为了确定食品对毒力的影响,将李斯特菌在发酵香肠和腌制熟火腿上生长后的侵袭 Caco-2 细胞的能力与在添加盐的 BHI 肉汤中生长的细菌进行了比较。样品在冷藏条件下储存长达 4 周。结果显示,在 10°C 的 BHI 肉汤或香肠中 7 天后,侵袭性没有差异(p > 0.05),而在火腿中孵育 2 和 4 周后,侵袭性略有增加(p < 0.05)与 BHI 肉汤相比。最重要的是,我们的结果表明,李斯特菌即使在冷藏温度下储存 4 周后,也能有效地侵袭 Caco-2 细胞。这对于该生物体在食品中的安全性评估非常重要,因为这些条件反映了即食食品在家庭冰箱中的储存情况。