Arachchi Geevika J Ganegama, Cruz Cristina D, Dias-Wanigasekera Beatrice M, McIntyre Lynn, Billington Craig, Hudson Andrew, Flint Steve H, Mutukumira Anthony N
Institute of Food, Nutrition and Human Health, Massey University, Albany Campus, North Shore City, Auckland, New Zealand.
The New Zealand Institute for Plant & Food Research (PFR) Limited, Mt Albert, Auckland, New Zealand.
Food Sci Technol Int. 2014 Dec;20(8):591-603. doi: 10.1177/1082013213497211. Epub 2013 Aug 1.
Listeria-infecting bacteriophages (listeriaphages) can be used to control Listeria monocytogenes in the food industry. However, the sensitivity of many of seafood-borne Listeria strains to phages has not been reported. This research investigated the host ranges of three listeriaphages (FWLLm1, FWLLm3 and FWLLm5) by the formation of lytic zones and plaques on host lawns and in vitro lysis kinetics of listeriaphage FWLLm3. The study also predicted the phage titres required to lyse host cells. The host ranges of the phages were determined using 50 L. monocytogenes strains, of which 48 were isolated from the seafood industry and two from clinical cases. Of the 50 strains, 36 were tested at 25 and 30 ℃ and the remainder (14) at 15 and 25 ℃. Based on the formation of either discrete plaques or lytic zones (host kill zones), the host ranges of FWLLm1, FWLLm3 and FWLLm5 were about 87%, 81% and 87%, respectively, at 25 ℃. Six L. monocytogenes strains from the seafood environment were insensitive to all three phages, while the other seafood strains (42) were phage-sensitive. The adsorption rate constant (k value) of listeriaphage FWLLm3 was between 1.2 × 10(-9) and 1.6 × 10(-9 )ml/min across four host strains in tryptic soy broth at 25 ℃. The cultures (at 3-4 log colony-forming unit (CFU/ml) were completely lysed (<1 log CFU/ml) when cultures were infected with FWLLm3 at > 8.7 log phage-forming units (PFU/ml) for 30 min. Re-growth of phage-infected cultures was not detected after 24 h. The effective empirical phage titre was similar to the calculated titre using a kinetic model. Results indicate the potential use of the three phages for controlling L. monocytogenes strains in seafood processing environments.
感染李斯特菌的噬菌体(李斯特菌噬菌体)可用于食品工业中控制单核细胞增生李斯特菌。然而,许多海鲜源李斯特菌菌株对噬菌体的敏感性尚未见报道。本研究通过在宿主菌苔上形成溶菌区和噬菌斑以及对李斯特菌噬菌体FWLLm3进行体外裂解动力学研究,调查了三种李斯特菌噬菌体(FWLLm1、FWLLm3和FWLLm5)的宿主范围。该研究还预测了裂解宿主细胞所需的噬菌体滴度。使用50株单核细胞增生李斯特菌菌株确定噬菌体的宿主范围,其中48株从海产品行业分离,2株来自临床病例。在这50株菌株中,36株在25℃和30℃下进行测试,其余14株在15℃和25℃下测试。基于离散噬菌斑或溶菌区(宿主杀伤区)的形成,在25℃时,FWLLm1、FWLLm3和FWLLm5的宿主范围分别约为87%、81%和87%。来自海产品环境的6株单核细胞增生李斯特菌菌株对所有三种噬菌体均不敏感,而其他海产品菌株(42株)对噬菌体敏感。在25℃的胰蛋白胨大豆肉汤中,李斯特菌噬菌体FWLLm3对四种宿主菌株的吸附速率常数(k值)在1.2×10⁻⁹至1.6×10⁻⁹毫升/分钟之间。当培养物用>8.7对数噬菌体形成单位(PFU/ml)的FWLLm3感染30分钟时,培养物(3-4对数菌落形成单位(CFU/ml))被完全裂解(<1对数CFU/ml)。24小时后未检测到噬菌体感染培养物的再生长。有效经验噬菌体滴度与使用动力学模型计算的滴度相似。结果表明这三种噬菌体在海产品加工环境中控制单核细胞增生李斯特菌菌株方面具有潜在用途。