Lactic acid bacteria and spoilage bacteria: Their interactions in Escherichia coli O157:H7 biofilms on food contact surfaces and implications for beef contamination

Abstract

This research explores the interaction between Shiga toxin-producing Escherichia coli (STEC) O157:H7 and bacteria species commonly found in beef processing environments, specifically Carnobacterium, Lactobacillus, Comamonas, Raoultella, and Pseudomonas. The study investigated how various environmental conditions impact the formation of biofilms and the ability of O157:H7 to transfer from multispecies biofilm onto beef surfaces. For this purpose, a mixture of lactic acid bacteria (LAB), spoilage bacteria (10⁶ CFU/mL), and E. coli O157 (10³ CFU/mL) were combined as follows: LAB (T1): Carnobacterium piscicola + Lactobacillus bulgaricus + O157:H7, an spoilage bacteria (T2): Comamonas koreensis + Raoultella terrigena + O157:H7, an spoilage bacteria (T3): Pseudomonas aeruginosa + C. koreensis strain + O157:H7 and only O157:H7 as control (T4). Multispecies biofilms were developed on thermoplastic polyurethane (TPU) and stainless steel (SS) coupons at 10 and 25°C for 6 days, washed and stored for 6, 30, and 60 days at wet (60%–90% RH) and dry (20%–50%, RH) conditions. To evaluate O157:H7 transfer, beef cubes (3 × 3 × 1 cm) were placed on the coupons, followed by a 50-g weight (7.35 kPa). The experiment was repeated three times in triplicate for each strain combination. Results demonstrate that biofilms formed at 10°C were generally weaker (less biomass) than those at 25°C. Regardless of temperature, more viable O157:H7 cells were transferred to beef from moist biofilms on TPU surfaces. At 25°C, T3 biofilm exhibited the lowest O157:H7 transfer to beef by 1.44 log₁₀ CFU/cm² (p < 0.01). At 10°C, none of the multispecies biofilm (T1–T3) affected the number of O157:H7 transfers to beef (p > 0.05). Notably, O157:H7 was not detected on food contact surfaces with 30 and 60-day-old dry biofilms (T1–T4). Through enrichment, E. coli O157:H7 was recovered from multispecies biofilms T1, T2, and T3. Findings from this study imply that multispecies biofilms contribute to the persistence of O157:H7 under dry conditions, regardless of temperature. These results underscore the intricate influence of multiple environmental factors—including surface type, biofilm age, humidity, temperature, and the presence of other bacterial species—on the risk of beef contamination facilitated by biofilms.