Trans-cinnamaldehyde nanoemulsion reduces Salmonella Enteritidis biofilm on steel and plastic surfaces and downregulates expression of biofilm associated genes

Abstract

Salmonella Enteritidis is a major poultry-associated foodborne pathogen that can form sanitizer-tolerant biofilms on various surfaces. The biofilm-forming capability of S. Enteritidis facilitates its survival on farm and food processing equipment. Conventional sanitization methods are not completely effective in killing S. Enteritidis biofilms. This study investigated the efficacy of a Generally Recognized as Safe phytochemical Trans-cinnamaldehyde (TC), and in its nanoemulsion form (TCNE), for inhibiting S. Enteritidis biofilm formation and inactivating mature biofilms developed on polystyrene and stainless-steel surfaces. Moreover, the effect of TC on Salmonella genes critical for biofilm formation was studied.

TCNE was prepared using a high energy sonication method with Tween 80. For biofilm inhibition assay, S. Enteritidis was allowed to form biofilms either in the presence or absence of sub-inhibitory concentration (SIC; 0.01 %) of TCNE at 25°C and the biofilm formed was quantified at 24-h intervals for 48 h. For the inactivation assay, S. Enteritidis biofilms developed at 25°C for 48 h were exposed to TCNE (0.5, 1 %) for 1, 5, and 15 min, and surviving S. Enteritidis in the biofilm were enumerated. SIC of TCNE inhibited S. Enteritidis biofilm by 45 % on polystyrene and 75 % on steel surface after 48 h at 25°C compared to control (P < 0.05). All TCNE treatments rapidly inactivated S. Enteritidis mature biofilm on polystyrene and steel surfaces (P < 0.05). The lower concentration of TCNE (0.5 %) reduced S. Enteritidis counts by 1.5 log CFU/ml as early as 1 min of exposure on both polystyrene and stainless-steel surfaces. After 15 min of exposure, TCNE at concentration of 0.5 or 1 % reduced S. Enteritidis count significantly by 4.5 log CFU or 6 log CFU/ml on polystyrene or stainless-steel surfaces. TC downregulated the expression of S. Enteritidis genes (hilA, hilC, flhD, csgA, csgD, sdiA) responsible for biofilm formation (P < 0.05). Results suggest that TCNE has potential as a natural disinfectant for controlling S. Enteritidis biofilms on common farm and food processing surfaces, such as plastic and steel.