Listeria monocytogenes biofilm-associated protein (BapL) may contribute to surface attachment of L. monocytogenes but is absent from many field isolates.

Listeria monocytogenes is a food-borne pathogen capable of adhering to a range of surfaces utilized within the food industry, including stainless steel. The factors required for the attachment of this ubiquitous organism to abiotic surfaces are still relatively unknown. In silico analysis of the L. monocytogenes EGD genome identified a putative cell wall-anchored protein (Lmo0435 [BapL]), which had similarity to proteins involved in biofilm formation by staphylococci. An insertion mutation was constructed in L. monocytogenes to determine the influence of this protein on attachment to abiotic surfaces. The results show that the protein may contribute to the surface adherence of strains that possess BapL, but it is not an essential requirement for all L. monocytogenes strains. Several BapL-negative field isolates demonstrated an ability to adhere to abiotic surfaces equivalent to that of BapL-positive strains. BapL is not required for the virulence of L. monocytogenes in mice.

Occurrence and persistence of Listeria spp. in the environment of ewe and cow's milk cheese dairies in Portugal unveiled by an integrated analysis of identification, typing and spatial-temporal mapping along production cycle.

Eight dairies, located in two distant geographic regions of Portugal, were screened along the production cycle in order to evaluate the presence and distribution of Listeria spp. in their environment. Three dairies in each region were positive for the presence of listeriae and 213 isolates were obtained. Based on an integrated analysis of RAPD fingerprints with three primers, molecular identification and genomic typing of isolates was performed followed by spatial and temporal mapping on dairy plants. The occurrence of Listeria species by region was noticeable different. Listeria monocytogenes prevailed in South Portugal dairies and L. innocua presented the highest occurrence in Azores, whereas L. seeligeri and L. ivanovii were detected in distinct regions. Dairies were at risk of contamination, from more than one source, whatever the stage in the production cycle and the surface materials used. For the three prevalent species, most of the genomic types were dairy and sampling time specific. Nonetheless, more than one type could be found in each dairy at a particular site and, in a few cases, even for different species. Some dairies also shared types, mainly for L. innocua and usually at the same stage of the production cycle. For L. monocytogenes, PCR serotyping was applied and 52% of genomic types were serotype 4b. An equal frequency of genomic types (24%) was found for serotypes 1/2b or 3b and 1/2a or 3a. The global pattern of types within a dairy is not constant, suggesting cycles of elimination and recontamination along the production cycle.

The resistance to detachment of dairy strains of Listeria monocytogenes from stainless steel by shear stress is related to the fluid dynamic characteristics of the location of isolation.

Strains of Listeria monocytogenes isolated from artisanal Portuguese cheese-making dairies were divided into two categories on the basis of the locations from which they were isolated: strains from dynamic locations were those that were habitually exposed to flowing liquids during the process of cheese-making, whereas those from static locations were rarely, if ever, exposed to the shear stresses generated by liquid flows. The strength of attachment to stainless steel discs of all of these strains was obtained using a radial flow chamber. Initial attachment strengths to stainless steel (after a 0.5 h contact time) of L. monocytogenes strains were greater for the 5 isolates from surfaces exposed to flow (dynamic isolates) than for most (3 out of 4) of those that were not (static isolates). After a 24 h contact time, attachment strength of all isolates reached similar levels. These results suggest that strains having high initial attachment strength are more likely to persist on surfaces exposed to flow than strains having low initial attachment strength. The numerical values of shear forces obtained could prove useful in the rational design of cleaning and decontamination procedures in food processing facilities.