Abstract
Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213) and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897) anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256). For the strictly anaerobic species, a high precision (relative standard deviation < 3.5%) was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells.
Highlights
Monitoring bacterial cell growth provides valuable information about their nutritional and energetic physiology, as well as an understanding of the survival and proliferation conditions of different species under different conditions (Koch, 2010)
We introduce here a new simple and portable setup for the rapid measurement of bacterial growth in culture tubes, by combining a commercial miniaturized spectrophotometer with a customized 3D printed tube holder
The pathogenic, facultative anaerobic bacteria Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213 were obtained from the General Hospital of Santo António – Centro Hospitalar do Porto (Porto, Portugal)
Summary
Monitoring bacterial cell growth provides valuable information about their nutritional and energetic physiology, as well as an understanding of the survival and proliferation conditions of different species under different conditions (Koch, 2010). The use of portable optical setups that could accommodate different formats of the sample holder (e.g., culture tubes, cuvettes, and flow cells), and that could be operated on bench and laminar flow chambers could contribute to faster, simpler, and safer monitoring of bacterial growth In this context, the advent of miniaturized spectrophotometers (Kantzas et al, 2009; Pena-Pereira et al, 2011) and 3D printers (Campbell et al, 2011) created the conditions for the customization of the measurement of OD for different microbiological applications. 3D printing is creating new opportunities to manufacture new customized laboratory hardware with simple design workflows and reduced costs (Pearce, 2014) Based on this background, we introduce here a new simple and portable setup for the rapid measurement of bacterial growth in culture tubes, by combining a commercial miniaturized spectrophotometer with a customized 3D printed tube holder. We applied this new manifold to the measurement of growth of pathogenic and rumen anaerobic bacteria, and compared the new approach to the classic measurement protocol based on benchtop spectrophotometers
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