Effect of Delmopinol Hydrochloride on the Prevention and Removal of Listeria monocytogenes and Salmonella enterica Stainless Steel-Adhered Biofilms.
Ewell, Ellen Sutton
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Bacterial biofilms attached to food contact surfaces are an ongoing concern for the food industry due to the resistance of bacteria within biofilms to detergents and sanitizers. Within food manufacturing facilities, stainless steel is a common food-contact surface in which microbial cell attachment and biofilm formation may occur. Identifying methods to prevent and remove biofilms during standard cleaning and sanitation practices could prove useful, as mature biofilms can release planktonic cells into an aqueous environment, causing continual low-level contamination. Dental studies involving delmopinol hydrochloride, a cationic surfactant, have found a preventative and dissociating affect on biofilms, where food applications have scarcely been researched. This study demonstrates the prevention and removal of Listeria monocytogenes 1/2a and S. enterica Agona biofilms on stainless steel with pre- and post-exposures of delmopinol hydrochloride. Stainless steel blanks (#304, 16 gauge, 2cm x 2cm, finish #4) were submerged in a 0.2% or 0.5% delmopinol solution before or after biofilm formation. Treatment times were 1, 5 or 10 minutes, whereas controls were not exposed to the delmopinol solution. Disinfected stainless steel blanks were spot-inoculated with 20µL of a 10⁹ CFU/mL liquid culture, and pre-exposed blanks were additionally submerged in delmopinol and dried prior to inoculation. Biofilms were exclusively formed on the finished and inoculated side by placing the surface face-down on TSA. After cell attachment and biofilm development for 24 hours at 25°C, blanks were rinsed with phosphate buffer. Post-exposed blanks were submerged in 0.2% or 0.5% delmopinol for 1, 5 or 10 minutes before all blanks were individually vortexed for 90 seconds to dislodge films. Bacterial populations were determined by surface plating onto TSA followed by incubation at 32°C for L. monocytogenes and 37°C for S. Agona for 48 hours. Treatments were in-duplicate and repeated three times for each microorganism. Pre-exposure of 0.2% delmopinol resulted in a significant decrease in L. monocytogenes concentration at 1, 5 and 10 minute exposures (P < 0.05). Pre-exposures with the 0.5% solution had no significant effect on L. monocytogenes biofilm populations (P > 0.05), whereas all post-exposures lead to a significant decline in biofilm concentrations (P < 0.0001). Post-exposures of 10 minutes exhibited a mean log₁₀ reduction of 5.59 and 6.40 log₁₀ for 0.2% and 0.5% delmopinol solutions, respectively. For S. Agona, 0.2% pre-exposure resulted in no significant log10 reduction (P > 0.05), while the 10 minute 0.5% pre-exposure exhibited a minimal reduction in bacterial growth (P < 0.05). Post-exposures of 10 minutes exhibited a mean log10 reduction of 7.65 and 7.75 log10 for 0.2% and 0.5% delmopinol solutions, respectively. For L. monocytogenes and S. Agona, post-exposure to delmopinol hydrochloride caused a notable log10 reduction. The removal effect of delmopinol on biofilms is significantly greater the preventative effect.
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