Laboratory of Aquatic Biomedicine, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742, Republic of Korea; Department of Bacteriology and Immunology, Medicum, Research Programs Unit, Immunobiology, University of Helsinki, PO Box 21, 00014 Helsinki, Finland.
Laboratory of Aquatic Biomedicine, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742, Republic of Korea.
Int J Food Microbiol. 2018 Apr 20;271:33-47. doi: 10.1016/j.ijfoodmicro.2018.02.007. Epub 2018 Feb 7.
Yersinia enterocolitica, the primary cause of yersiniosis, is one of the most important foodborne pathogens globally and is associated with the consumption of raw contaminated pork. In the current study, four virulent bacteriophages (phages), one of Podoviridae (fHe-Yen3-01) and three of Myoviridae (fHe-Yen9-01, fHe-Yen9-02, and fHe-Yen9-03), capable of infecting Y. enterocolitica were isolated and characterized. fHe-Yen9-01 had the broadest host range (61.3% of strains, 65/106). It demonstrated a latent period of 35 min and a burst size of 33 plaque-forming units/cell, and was found to have a genome of 167,773 bp with 34.79% GC content. To evaluate the effectiveness of phage fHe-Yen9-01 against Y. enterocolitica O:9 strain Ruokola/71, we designed an experimental model of the food market environment. Phage treatment after bacterial inoculation of food samples, including raw pork (4 °C, 72 h), ready-to-eat pork (26 °C, 12 h), and milk (4 °C, 72 h), prevented bacterial growth throughout the experiments, with counts decreasing by 1-3 logs from the original levels of 2-4 × 10 CFU/g or ml. Similarly, when artificially contaminated kitchen utensils, such as wooden and plastic cutting boards and knives, and artificial hands, were treated with phages for 2 h, bacterial growth was effectively inhibited, with counts decreasing by 1-2 logs from the original levels of ca 10 CFU/cm or ml. To the best of our knowledge, this is the first report of the successful application of phages for the control of Y. enterocolitica growth in food and on kitchen utensils.
产单核细胞李斯特菌是食源性疾病病原菌之一,主要通过食用被污染的生猪肉而感染。本研究分离鉴定了一株肠侵袭性大肠埃希菌噬菌体 fHe-Yen3-01(Podoviridae)和三株肠侵袭性大肠埃希菌噬菌体 fHe-Yen9-01、fHe-Yen9-02、fHe-Yen9-03(Myoviridae)。fHe-Yen9-01 宿主谱最广(61.3%,65/106),潜伏期为 35min,爆发量为 33 个噬菌斑/细胞,基因组大小为 167773bp,GC 含量为 34.79%。为评估噬菌体 fHe-Yen9-01 对食源性 O:9 型产单核细胞李斯特菌 Ruokola/71 的防治效果,构建了食品市场环境的模拟实验模型。在接种食源性样品(4°C,72h 的生猪肉;26°C,12h 的即食猪肉;4°C,72h 的牛奶)后进行噬菌体处理,噬菌体 fHe-Yen9-01 可有效抑制细菌生长,与对照组相比,实验组的菌数减少 1-3 个对数级,原始菌数为 2-4×10⁵CFU/g 或 ml。此外,将噬菌体处理过的厨房用具(木制和塑料案板、刀具,以及人工手)进行人工污染,当作用 2h 后,同样能有效抑制细菌生长,与对照组相比,实验组的菌数减少 1-2 个对数级,原始菌数为 10⁵CFU/cm 或 ml 左右。据我们所知,这是首次报道噬菌体在食品和厨房用具中成功控制产单核细胞李斯特菌生长的应用。