Produce Safety and Microbiology Research Unit, U.S. Department of Agriculture, Agricultural Research Service, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA.
Produce Safety and Microbiology Research Unit, U.S. Department of Agriculture, Agricultural Research Service, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA.
Cryobiology. 2020 Oct;96:85-91. doi: 10.1016/j.cryobiol.2020.07.012. Epub 2020 Aug 1.
Lyophilization is commonly used to effectively preserve the stability of bacteriophages (phages) in long-term storage. However, information regarding the lyophilization of phages specific to Shiga toxin-producing Escherichia coli (STEC) strains is scarce. The objective of this study was to determine the effects of lyophilization with different cryoprotectants (sucrose and trehalose) and concentrations (0.1 M and 0.5 M) on the stability of seven lytic phages specific to STEC O157 and top 6 non-O157 strains during 6-month storage at -80 °C. The titers of lyophilized phages specific to STEC O26 (S1 O26) and STEC O121 (Pr121lvw) did not exhibit significant reduction after 6-month storage regardless of the use of cryoprotectants. Phages lytic against STEC O103 (Ro103C3lw) and STEC O145 (Ro145clw) with 0.1 M sucrose retained similar titers after lyophilization and frozen storage for 6 months (P > 0.05). Despite subtle differences, these results indicated that most of the selected phages had similar titer retention with the same cryoprotectants. Additionally, lytic activities of the phages against their primary hosts were not affected after lyophilization and 6-month frozen storage. Moreover, no detectable damage was observed on the lyophilized phage structures. These findings provide valuable insight into the use of lyophilization to preserve phages lytic against STEC strains.
冷冻干燥通常用于有效保存噬菌体(phages)在长期储存中的稳定性。然而,关于特定产志贺毒素大肠杆菌(STEC)菌株噬菌体的冷冻干燥信息却很少。本研究的目的是确定使用不同的保护剂(蔗糖和海藻糖)和浓度(0.1 M 和 0.5 M)对 7 种针对 STEC O157 和前 6 种非 O157 菌株的溶菌噬菌体在-80°C 下储存 6 个月的稳定性的影响。在 6 个月的储存期内,无论是否使用保护剂,针对 STEC O26(S1 O26)和 STEC O121(Pr121lvw)的冷冻干燥噬菌体的滴度均未显著降低。对于含有 0.1 M 蔗糖的针对 STEC O103(Ro103C3lw)和 STEC O145(Ro145clw)的噬菌体,在冷冻干燥和 6 个月的冷冻储存后保留了相似的滴度(P > 0.05)。尽管存在细微差异,但这些结果表明,大多数选定的噬菌体在相同的保护剂下具有相似的滴度保持能力。此外,在冷冻干燥和 6 个月的冷冻储存后,噬菌体对其主要宿主的溶菌活性不受影响。此外,在冷冻干燥的噬菌体结构上未观察到可检测到的损伤。这些发现为使用冷冻干燥保存针对 STEC 菌株的溶菌噬菌体提供了有价值的见解。