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''Brettanomyces'' has the ability to form a [[Quality_Assurance#Biofilms|biofilm]]. Biofilm formation is a survival mechanism induced by stress whereby the cells adhere to non-living surfaces such as plastic and stainless steel. After adhesion to the surface, the cells produce a protective layer of proteins and polysaccharides that help protect the organism from cleaning and sanitizing agents.
Joseph et al. (2007) tested 36 wine strains of ''B. bruxellensis'' for biofilm formation over a 10 day period. They found that just under half of the strains formed a biofilm, and about half of those formed considerable and consistent biofilms throughout the tests. Almost all strains tested (95%) adhered to a surface with 0.1% glucose within 6 hours of contact (the same conditions that get ''Saccharomyces cerevisiae'' to adhere to a surface; longer contact with surfaces and higher residual sugar could encourage ''Brettanomyces'' to adhere more readily to surfaces). A juice-based growth media in the range range of 2 - 4.5 pH was tested for biofilm formation and 3-4 for cell adhesion to a surface, and for most of the strains they formed stronger biofilms and adhered better in the higher pH growth media (4.5 pH being the highest tested). Under a pH of 3.5 significantly dropped biofilm formation and adherence, indicating that something about pH affects the cells ability to attach themselves. The researchers concluded that winemakers should keep wine in the lower end of the pH range (3.5). Six different types of cleaners were tested to see how well they removed the biofilms: keytones + surfactant detergent, quaternary ammonia + surfactant detergent, sodium hydroxide (caustic soda), sodium carbonate (soda ash), sodium hydroxide + surfactant (alkaline detergent), and chlorine (sanitizer, not a detergent). They found that only caustic soda was consistently efficient at removing the biofilm. The chlorine, while it did not remove the biofilm, still killed all of the ''Brettanomyces'' cells, and it was presumed that the other cleaners might have killed the ''Brettanomyces'', but that was not tested for. They also tested to see if cells that were adhered to a surface could be cleaned. Again, the caustic soda performed consistently the best, but the ammonia + surfactant cleaner and the quaternary ammonia + surfactant detergent also effectively removed adhered cells. The other cleaners varied in how well they removed adhered cells from a surface <ref>[https://www.researchgate.net/publication/235411588_Adhesion_and_biofilm_production_by_wine_isolates_of_Brettanomyces_bruxellensis Adhesion and biofilm production by wine isolates of Brettanomyces bruxellensis. C. M. Lucy Joseph, Gagandeep Renuka Kumar, Gagandeep Renuka Kumar, Edward Su, Linda F Bisson. 2006. American Journal of Enology and Viticulture 58(3):373-378.]</ref>.
Dimopoulou et al. (2019) studied ''Brettanomyces bruxellensis'' biofilms from each of the genetic branches of ''B. bruxellensis''. They found that for the wine strains biofilms formed more readily when grown in wine must rather than YPD media; however, the beer strains grew biofilms equally well in wine must and YPD media. The biofilms contained a large portion of saturated fatty acids and a smaller portion of monounsatured fatty acids. The amount of exopolysaccharide produced varied widely across the strains tested per cell population, with some wine strains producing little EPS (40 mg/L/OD), beer strains producing moderate amounts, and the other wine group producing a high amount (100 mg/L/OD). Additionally, the different strains displayed a varying degree of negative cell wall charges, with the beer and tequila strains being more negatively charged than wine strains, which could help them adhere to surfaces and form biofilm <ref>[https://europepmc.org/article/PPR/PPR73221?singleResult=true Dimopoulou M., Renault M., Dols-Lafargue M., Albertin-Leguay W., Herry J., Bellon-Fontaine M., Masneuf-Pomarede I. 2019. DOI: 10.1101/579144.]</ref>.