ABSTRACT
Introduction: Staphylococcus aureus is an important pathogen that can form biofilms on food contact surfaces (FCS) in the dairy industry, posing a serious food safety, and quality concern. Biofilm is a complex system, influenced by nutritional-related factors that regulate the synthesis of the components of the biofilm matrix. This study determines the prevalence of biofilm-associated genes and evaluates the development under different growth conditions and compositions of biofilms produced by S. aureus. Methods: Biofilms were developed in TSB, TSBG, TSBNaCl, and TSBGNaCl on stainless-steel (SS), with enumeration at 24 and 192 h visualized by epifluorescence and scanning electron microscopy (SEM). The composition of biofilms was determined using enzymatic and chemical treatments and confocal laser scanning microscopy (CLSM). Results and discussion: A total of 84 S. aureus (SA1-SA84) strains were collected from 293 dairy industry FCS (FCS-stainless steel [n = 183] and FCS-polypropylene [n = 110]) for this study. The isolates harbored the genes sigB (66%), sar (53%), agrD (52%), clfB/clfA (38%), fnbA/fnbB (20%), and bap (9.5%). 99. In particular, the biofilm formed by bap-positive S. aureus onto SS showed a high cell density in all culture media at 192 h in comparison with the biofilms formed at 24 h (p < 0.05). Epifluorescence microscopy and SEM revealed the metabolically active cells and the different stages of biofilm formation. CLSM analysis detected extracellular polymeric of S. aureus biofilms on SS, such as eDNA, proteins, and polysaccharides. Finally, the level of detachment on being treated with DNase I (44.7%) and NaIO 4(42.4%) was greater in the biofilms developed in TSB compared to culture medium supplemented with NaCl at 24 h; however, there was no significant difference when the culture medium was supplemented with glucose. In addition, after treatment with proteinase K, there was a lower level of biomass detachment (17.7%) of the biofilm developed in TSBNaCl (p < 0.05 at 24 h) compared to that in TSB, TSBG, and TSBGNaCl (33.6, 36.9, and 37.8%, respectively). These results represent a deep insight into the composition of S. aureus biofilms present in the dairy industry, which promotes the development of more efficient composition-specific disinfection strategies.
ABSTRACT
Sprouts are vehicles of foodborne diseases caused by pathogens such as Salmonella. The aim of this study was to evaluate thermal and chemical treatments applied as a hurdle approach to reduce Salmonella in alfalfa (Medicago sativa) and broccoli (Brassica oleracea var. italica) seeds before and during their germination. Seeds, inoculated and then dried at 55°C for 48 h, were subjected to a chemical treatment and a thermal shock with (i) 75 mM caprylic acid at 70°C for 5 s, (ii) 0.04% CaO at 70°C for 5 s, or (iii) 1% H2O2 at 70°C for 5 s. After each treatment, seeds were immersed in water at 3°C for 5 s. Next, the imbibition process was carried out with 0.016% H2O2 at pH 3.0. Finally, the seeds were transferred to a rotary drum-type germinator and were sprayed with the same chemical solution that was applied before the imbibition process, for 20 s at intervals of 5 min for 40 min at 3 rpm. All chemical treatments reduced Salmonella at least 5 log CFU/g on both seeds. Germination rates between 90 and 93% were obtained after application of thermal and chemical treatments. Salmonella was not detected after the imbibition stage when caprylic acid and H2O2 treatments were applied. However, during the germination process of both seeds, Salmonella counts of >6 log CFU/g were obtained despite all treatments being applied at different stages of the sprouting process. These results demonstrated that thermal and chemical treatments used as a hurdle approach to control Salmonella on alfalfa and broccoli seeds significantly reduced the pathogen concentration on seeds >5 log but were ineffective to eliminate Salmonella and to control its growth during the sprouting process. The production of safe sprouts continues to be a major challenge for industry.
