Elucidating the biofilm formation process, microstructure and functional gene expression of Listeria monocytogenes in beef juice

Abstract

Listeria monocytogenes biofilm is recognized as a frequent cross-contamination source in the food industry, with raw beef and beef products as common food reservoirs. L. monocytogenes sequence types 9 (ST9) and ST8 are frequently isolated in meats and meat processing environment. In this study, beef juice was selected and compared to a laboratory medium (tryptone soy broth with 0.6 % yeast extract, TSB-YE). The purpose of this work was to investigate the effect of beef juice on the biofilm formation of ST9 and ST8 strains, including biofilm microstructure and modelling the biofilm formation process. Then the expression of biofilm functional genes in two culture media was also investigated. L. monocytogenes ST9 and ST8 can form a dense three-dimensional structure biofilm with multilayers of cells in beef juice after 48 h of incubation, but both strains formed a monolayer biofilm structure in TSB-YE. The ST9 strain developed more sessile cells on the stainless-steel surfaces than the ST8 strain under the same culture conditions. The Logistic model showed a good fit for with the biofilm formation process, and the estimated model parameters in beef juice and TSB-YE were considerably different. Under the same conditions, the maximum specific biofilm formation rate (μ_max) in beef juice was higher than that in TSB-YE. This indicated that beef juice can facilitate the biofilm formation of L. monocytogenes, suggesting that the particles in beef juice act as a surface conditioner to support attachment. However, the maximum counts of L. monocytogenes biofilm formed on stainless steel coupon (Y_max) in beef juice was smaller than that in TSB-YE. The ST9 strain exhibited a stronger biofilm formation ability than the ST8 strain, and this was consistent with the scanning electron microscopy images. In the corresponding culture suspensions, the number of adherent cells increases with the number of planktonic cells. Moreover, the expression of biofilm functional genes was significantly different in the two culture media. Compared to biofilm cultured in TSB-YE, the expression of the agrA gene of biofilm in beef juice was significantly down-regulated for both the ST9 and the ST8 strains, and the expression of the inlB and the actA genes were dramatically up-regulated for the ST8 strain. Our results suggested that beef juice promotes biofilm formation of L. monocytogenes in meat processing and provide new insights into controlling biofilm.