Features of Bioluminescence Dynamics of Photobacterium phosphoreum IMV B-7071

Gromozova, O. M., Martynyuk, V. S., Artemenko, O. Y., Hretskyi, I. O., Mulec, J., & Tseyslyer, Y. V. (2024). Features of Bioluminescence Dynamics of Photobacterium phosphoreum IMV B-7071. Mikrobiolohichnyi Zhurnal, 86(4), 3–18. doi:10.15407/microbiolj86.04.003

The problem of the prolonged and stable intensity of bioluminescent signals is relevant in the development of any test systems that use biological objects. The aim of this work was to study the features of bioluminescence dynamics of Photobacterium phosphoreum IMV B-7071 in a liquid and on different stationary media. Methods. Bioluminescence studies were performed in liquid, agarose, and cellulose-cotton media. Bacterial suspensions were cultivated at 21°C in the mediums with standard composition. We studied both the background glow and its dynamics under conditions of mixing a liquid medium. Bioluminescence was recorded using digital photography with subsequent image processing of the samples. The measurements of luminescence were made by digital photo or video recording using Olympus digital camera SP560UZ, CANON 700D, and mobile device camera Samsung Galaxy 9 Note with specialized applications for mobile devices "Colorimeter (Lab Tools Apps)" and Camera Color Counter (Keuwsoft) at maximum light sensitivity in the automatic white balance mode at a fixed distance from the sample. Image processing was carried out using ImageJ and Origin Pro. Spectra of bacterial luminescence and its dynamics over time were measured using an LOMO MDR-23 spectrometer in the range of 200–750 nm. Results. The results of the study prove that in aqueous or solid agar and also on cellulose cotton medium, the intensity of bioluminescence of P. phosphoreum gradually increases, reaching a maximum within approximately 2 days, after which it slowly fades. It was established that the bioluminescence of photobacterium P. phosphoreum is a non-stationary process and has characteristic features of temporal dynamics associated with both the dynamics of the oxygen concentration in the environment of bacterial suspensions and the dynamics of the bacterial population density. Analysis of the luminescence spectra of bacteria shows that luminescence occurs mainly in the blue and green regions of the spectrum with luminescence maxima in the range of 460–520 nm, but the ultra-weak glow is also registered in the UV and red spectral ranges. The variability of photobacterial luminescence spectra over time in the spectral ranges of the main luminophores causes color fluctuations between the blue and green ranges. Conclusions. The key parameters of multi-day background and short-term induced bioluminescence dynamics of photobacteria in different environments were clarified, and the certain variability of the spectral characteristics of luminescent radiation over time was shown. The revealed features of the dynamics of the bioluminescence of P. phosphoreum must be taken into account in practical application to assess the toxicity of substances of various nature, as well as in environmental monitoring.