Nestlé Research Center, Nestec Ltd, Lausanne, Switzerland.
Microb Biotechnol. 2008 Nov;1(6):532-43. doi: 10.1111/j.1751-7915.2008.00058.x.
Enterobacter sakazakii (Cronobacter spp.) is an opportunistic pathogen, which can cause rare, but life-threatening infections in neonates and infants through feeding of a contaminated milk formula. We isolated 67 phages from environmental samples and tested their lytic host range on a representative collection of 40 E. sakazakii strains. A cocktail of five phages prevented the outgrowth of 35 out of 40 test strains in artificially contaminated infant formula. Two E. sakazakii phages represented prolate head Myoviridae. Molecular tests identified them as close relatives of Escherichia coli phage T4. The remaining three phages represented isometric head Myoviridae with large genome size of 140 and 200 kb, respectively, which belonged to two different DNA hybridization groups. A high dose of 10(8) pfu ml(-1) of phage could effectively sterilize a broth contaminated with both high and low pathogen counts (10(6) and 10(2) cfu ml(-1)). In contrast, broth inoculated with 10(4) phage and 10(2) bacteria per ml first showed normal bacterial growth until reaching a cell titre of 10(5) cfu ml(-1). Only when crossing this threshold, phage replication started, but it could not reduce the contamination level below 100 cfu ml(-1). Phages could be produced with titres of 10(10) pfu ml(-1) in broth culture, but they were not stable upon freeze-drying. Addition of trehalose or milk formula stabilized the phage preparation, which then showed excellent storage stability even at elevated temperature.
阪崎肠杆菌(克罗诺杆菌属)是一种机会致病菌,通过喂食受污染的配方奶粉,可导致新生儿和婴儿发生罕见但危及生命的感染。我们从环境样本中分离出 67 株噬菌体,并在代表 40 株阪崎肠杆菌菌株的集合上测试了它们的裂解宿主范围。五株噬菌体的混合物可防止 40 株测试菌株中的 35 株在人工污染的婴儿配方奶粉中生长。两种阪崎肠杆菌噬菌体代表长头肌尾噬菌体科。分子测试将它们鉴定为大肠杆菌噬菌体 T4 的近亲。其余三种噬菌体代表等径头肌尾噬菌体科,基因组大小分别为 140 和 200kb,分别属于两个不同的 DNA 杂交群。高剂量(10^8 pfu/ml)的噬菌体可有效杀灭高和低病原体计数(10^6 和 10^2 cfu/ml)污染的肉汤。相比之下,用 10^4 噬菌体和 10^2 个细菌/ml 接种的肉汤首先显示出正常的细菌生长,直到细胞浓度达到 10^5 cfu/ml。只有当超过这个阈值时,噬菌体才开始复制,但它无法将污染水平降低到 100cfu/ml 以下。噬菌体在肉汤培养物中的产量可达 10^10 pfu/ml,但在冻干时不稳定。添加海藻糖或配方奶粉可稳定噬菌体制剂,即使在高温下,也能显示出极好的储存稳定性。