Abstract
Biofilms pose a serious problem to the food industry due to their high resistance to stressing conditions, including antimicrobials and disinfectants. Therefore, it is of vital importance to have methods that allow us to determine and quantify the cells of which biofilms are composed in order to determine the effectiveness of cleaning and disinfection treatments. In this chapter, we suggest two techniques, the plate counting technique and the crystal violet staining technique, as two possible indirect methods to determine in vitro biofilm mass. To overcome individual limitations, such as the plate counting technique’s disregard of the amount and localization of biomass on surfaces, or the crystal violet staining technique’s failure to differentiate between living and dead cells, we propose their combined use in order to obtain complete, valuable information on the behavior of microbial biofilms.
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Acknowledgments
This research was supported by the Agencia Estatal de Investigación (Spain) (PGC2018-093789-B-I00), by the European Social Fund, by the Aragonese Office of Science, University and Knowledge Society (research contract to N.M.), and by the Spanish Ministry of Science and Innovation (FPU17/02441 to E.P.; FPU15/02703 to D.B.).
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Gascón, E., Merino, N., Pagán, E., Berdejo, D., Pagán, R., García-Gonzalo, D. (2021). Assessment of In Vitro Biofilms by Plate Count and Crystal Violet Staining: Is One Technique Enough?. In: Magnani, M. (eds) Detection and Enumeration of Bacteria, Yeast, Viruses, and Protozoan in Foods and Freshwater. Methods and Protocols in Food Science . Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1932-2_6
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