Muthuraman SPalmer JFlint S2025-12-042026-04Muthuraman S, Palmer J, Flint S. (2026). Sequential treatment of psychrotrophic pseudomonad biofilms with sodium hydroxide and commercial enzyme cleaners. Food Control. 182.0956-7135https://mro.massey.ac.nz/handle/10179/73907Pseudomonads are psychrotrophic spoilage bacteria that can form biofilms at the air-liquid interface. Food processing utensils and equipment often facilitate the air-liquid interface biofilm formation. Pseudomonads produce thermostable enzymes and pigments that affect the organoleptic quality of perishable food products. In this study, Pseudomonas lundensis, Pseudomonas cedrina were allowed to form biofilms at 4 °C under continuous flow of nutrients in a CDC reactor (CBR 90; Biosurface Technologies, USA). The mature biofilms were treated with commercial enzyme cleaners, EnduroZyme (protease), DualZyme (protease and lipase), and TriZyme (protease, amylase, and cellulase). The dispersion with EnduroZyme was significantly (p < 0.05) higher than the other enzyme cleaners. Then the biofilms were treated with hot water and sodium hydroxide, and enzyme cleaners (sequential treatment). The cell counts after sodium hydroxide + Enzyme cleaners were below the detection limit. The microscopic observations with epifluorescence microscopy showed that the coupons had less fluorescence after the sequential treatment. FTIR observations showed that the extracellular polymeric substances (EPS) isolated after sodium hydroxide + enzyme cleaners differed from the untreated and sodium hydroxide-only-treated EPS. Biofilm regrowth was significantly (p < 0.05) lower in the biofilms treated with sodium hydroxide + EnduroZyme compared to acid-treated control coupons. The sequential treatment with sodium hydroxide and enzyme cleaners reduced the biofilm footprints, representing a better clean than enzyme treatment alone or sodium hydroxide-only cleaning.CC BY 4.0(c) 2025 The Author/shttps://creativecommons.org/licenses/by/4.0/PsychrotrophicAir-liquid interfacePseudomonadsDispersionEnzymesJournal article10.1016/j.foodcont.2025.1118581873-7129journal-articlehttps://orcid.org/0000-0002-1027-6241111858S0956713525007273