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  • Title: CO2-Based micro-respirometry for measuring bacterial load under aerobic and anaerobic conditions.
    Author: McDonnell L, Yusufu D, Mills A.
    Journal: Analyst; 2024 Nov 18; 149(23):5582-5590. PubMed ID: 39351844.
    Abstract:
    The bacterial load (BL), or total viable count, of aerobes can be measured using micro-respirometry, %O2-μR, in which the consumption of dissolved O2 is monitored with respect to incubation time, t. In %O2-μR the 'bioreactor' often comprises a canonical plastic tube with a small %O2 sensor; it is simple, fast and accurate and used in automated, commercial instruments for measuring BL. Here we show that it is also possible to measure BL using a new form of micro-respirometry, %CO2-μR, in which the production of CO2 in the growth medium is monitored. In %CO2-μR, the 'bioreactor' is the same as that used in %O2-μR, but with a small 3D printed, colour-based %CO2 indicator set in its base and its apparent absorbance, A', is measured at any t, as it is related to the %CO2 dissolved in the inoculated growth medium. Under aerobic conditions, different inoculations of the facultative anaerobe, E. coli, of different concentrations (101-108 colony forming units (CFU) per mL) are used to generate a series of A' vs. t profiles, and a straight-line calibration curve. Statistical comparative analysis of the results generated in the above %CO2-μR study, to those generated for the same system but using a commercial %O2-μR system, is used to demonstrate method equivalence. A study of the same system, under anaerobic conditions, using %CO2-μR, shows that %CO2-μR is suitable for measuring the BL of anaerobes. The potential of %CO2-μR for measuring the bacterial load of CO2-generating aerobes and anaerobes is discussed briefly.
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