Comparative evaluation of adhesion and biofilm formation of different Listeria monocytogenes strains

Thirteen Listeria monocytogenes strains were used to grow biofilms on glass surfaces in static conditions at 37 degrees C for up to 4 days. After the initial 3-h adhesion and in subsequent 1-day intervals, cell numbers were determined using standard plate count after swabbing the cells from the glass surface. The three-dimensional structure of in situ biofilms was determined by confocal scanning laser microscopy (CSLM). After 3 h incubation, bacterial cells for all 13 strains of L. monocytogenes were found attached to glass slides and all strains formed biofilms within 24 h. The strains varied significantly in their ability to adhere to the surface and significant differences for cell numbers after 24 h biofilm growth were found. Cell counts in biofilms formed by five L. monocytogenes strains were monitored over 4 days. The counts increased for the first 2 days reaching 10(5) cfu/cm2, except for L. monocytogenes 7148 (10(4) cfu/cm2). After 2 days, cell counts remained at 10(5) cfu/cm2 for four strains (tested on days 3 and 4), while L. monocytogenes 7148 continued to grow and reached 10(5) cfu/cm2 on day 4. This difference in biofilm growth was not related to variations in growth rates of planktonic cells suggesting that growth behaviour of Listeria in biofilms may be different from their planktonic growth. CSLM revealed that the biofilms grown under static conditions consisted of two distinct layers with 0.5 log10 higher cell numbers in the bottom layer as compared to the upper layer.