Simple, reusable, solid-state system for measuring total (aerobic) viable count, TVC, using the micro-respirometry method (µRM)
Simple, reusable, solid-state system for measuring total (aerobic) viable count, TVC, using the micro-respirometry method (µRM)
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简单、可重复使用的固态系统，用于使用微呼吸测量法 (µRM) 测量总（有氧）活菌数 TVC
DOI：
10.1016/j.snb.2024.135435
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Watson M
中科院分区：
文献类型：
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作者：
Watson M
液体微量呼吸测定法，Liq-μRM，是一种流行的、更快速的测量需氧菌总活菌计数的替代方法，即，TVC，单位：菌落形成单位（CFU）/mL，以传统的、耗时的平板计数法。Liq-μRM基于监测作为孵育时间函数的液体生长培养基中溶解O2的消耗。然而，在Liq-μRM中，由于搅拌问题，O2指示剂难以制作，不可重复使用，并且不太适合在孔板中使用。在这项工作中，一种新的，固态微量呼吸法，即，SS-μRM基于固体而非液体生长培养基进行描述，其中使用3D打印的O2指示剂监测顶部空间中O2的消耗，该指示剂设置在含有固体生长培养基的小培养皿或孔板的盖子中。在小培养皿中比较了SS-μRM与商用Liq-μRM系统的性能，使用大肠杆菌，即，E.大肠杆菌，作为试验细菌，在101- 108 CFU/mL范围内，并证明具有等效性。同样的系统被用来测量铜绿假单胞菌和阴沟肠杆菌的TVC。大肠杆菌在101- 108 CFU/mL范围内。当用于测量E.在孔板中，SS-μRM对搅拌不敏感，而Liq-μRM对搅拌不敏感。讨论了SS-μRM解决有关Liq-μRM的主要问题的能力。
Liquidmicro-respirometry, Liq-μRM, is a popular, more rapid alternative method for measuring Total Viable Count of aerobes, i.e., TVC, units: colony forming units (CFU)/mL, to the traditional, time-consuming, plate counting method. Liq-μRM is based on monitoring the consumption of dissolved O2in aliquidgrowth medium as a function of incubation time. However, in Liq-μRM, the O2indicator is difficult to make, non-reusable and not well suited for use in well-plates, due to agitation issues. In this work a new, Solid-State micro-respirometry method, i.e., SS-μRM, is described based on asolid, rather thanliquid, growth medium, in which the consumption of O2in the headspace is monitored, using a 3D printed O2indicator set in the lid of a small Petri dish or well plate containing the solid growth medium. The performance of the SS-μRM in a small Petri dish is compared with that of a commercial Liq-μRM system, usingEscherichia coli, i.e.,E. coli, as the test bacterium over the range 101– 108CFU/mL and demonstrated to be equivalent. The same system is used to measure the TVC ofPseudomonas aeruginosa,P. aeruginosa,andEnterobacter cloacae,E. cloacae,over the range 101– 108CFU/mL. When used to measure the TVC ofE. coli, in a well plate, SS-μRM is shown to be insensitive to agitation, in contrast to Liq-μRM. The ability of the SS-μRM to address the main concerns regarding Liq-μRM are discussed.