An experimental model was proposed to study biofilm formation by Listeria monocytogenes ATCC 19117 on AISI 304 (#4) stainless steel surface and biotransfer potential during this process. In this model, biofilm formation was conducted on the surface of stainless steel coupons, set on a stainless steel base with 4 divisions, each one supporting 21 coupons. Trypic Soy Broth was used as bacterial growth substrate, with incubation at 37 ºC and stirring of 50 rpm. The number of adhered cells was determined after 3, 48, 96, 144, 192 and 240 hours of biofilm formation and biotransfer potential from 96 hours. Stainless steel coupons were submitted to Scanning Electron Microscopy (SEM) after 3, 144 and 240 hours. Based on the number of adhered cells and SEM, it was observed that L. monocytogenes adhered rapidly to the stainless steel surface, with mature biofilm being formed after 240 hours. The biotransfer potential of bacterium to substrate occurred at all the stages analyzed. The rapid capacity of adhesion to surface, combined with biotransfer potential throughout the biofilm formation stages, make L. monocytogenes a potential risk to the food industry. Both the experimental model developed and the methodology used were efficient in the study of biofilm formation by L. monocytogenes on stainless steel surface and biotransfer potential.
Essential oils are plant secondary metabolites commonly used in traditional medicine to treat infectious diseases. Along with their compounds, oils can contribute to development of new antimicrobial/antibiofilm products. Our study evaluated antibacterial activity of essential oils and their major compounds on Escherichia coli and Staphylococcus aureus planktonic cells and anti-biofilm activity. The effect of essential oils and their major compounds on biofilm and planktonic cells was assessed by quantifying the number of viable cells (CFU). Biomass quantification (absorbance = OD570nm) was also performed to evaluate anti-biofilm activity. Planktonic cells were more susceptible to the action of agents. Escherichia coli was reduced by 100% with cinnamon and palmarosa oil. The treatment showed an interesting anti-biofilm activity, whereas green tea essential oil and its major compound, terpinen-4-ol, yielded less effective results. Reduction of viable cells in biofilm biomass was significant. Although our research is one of the first experiments in anti-biofilm activity of essential oils and their compounds against Escherichia coli and Staphylococcus aureus, pharmacological data confirm that the materials used in the trial do not pose health risk. Thus, essential oils and their compounds can be safely used in research to identify new antibacterial and anti-biofilm products against pathogenic bacteria.
The antimicrobial activity of linalool, citral, eugenol and thymol was determined in growth studies of both planktonic (PC) and biofilm cells (BC) Shigella flexneri. These components were evaluated either in isolation or in combinations using a sequential experimental strategy with Plackett & Burman and central composite rotational designs totaling 47 treatments. The minimum inhibitory concentration for PC was 0.125% (v v−1) for linalool and 0.5% (v v−1) for citral, eugenol and thymol. The biofilm minimum bactericidal concentration was 3 and 1% (v v−1) for linalool and citral, respectively, and 2% (v v−1) for eugenol and thymol. In the mixtures, the minimum concentrations in the efficient assays for PC growth inhibition were 0.0003, 0.0443 and 0.0443% (v v−1), for linalool, citral and thymol, respectively. In the BC, only two assays with concentrations of 0.0558, 0.0558 and 0.319% (v v−1) and 0.035, 0.035 and 0.3999% (v v−1) for linalool, citral and thymol, respectively, inhibited Shigella growth. Synergism was observed among the components, where PC and BC growth inhibition occurred at lower concentrations than those noted individually. The bactericidal effect of the components in microplate was different from the observed in stain steel coupons. Therefore, the obtained model can describe and predict the PC count of S. flexneri in medium with the tested compounds and they could be an alternative for the use in microbiological control in food industry.
In this study, the antioxidant, antimicrobial, mechanical, optical, and barrier attributes of Solanum lycocarpum starch bio-based edible films incorporated with a phenolic extract from jaboticaba peel were investigated. Aiming to determine the effect of the polymers and the phenolic extract on the properties of the films, a three-factor simplex-lattice design was employed, and the formulation optimization was based on the produced films’ antioxidant potential. The optimized formulation of the starch-PEJP film showed a reddish-pink color with no cracks or bubbles and 91% antioxidant activity against DPPH radical. The optimized starch-PEJP film showed good transparency properties and a potent UV-blocking action, presenting color variation as a function of the pH values. The optimized film was also considerably resistant and highly flexible, showing a water vapor permeability of 3.28 × 10−6 g m−1 h−1 Pa−1. The microbial permeation test and antimicrobial evaluation demonstrated that the optimized starch-PEJP film avoided microbial contamination and was potent in reducing the growth of Escherichia coli, Staphylococcus aureus, and Salmonella spp. In summary, the active starch-PEJP film showed great potential as an environmentally friendly and halochromic material, presenting antioxidant and antimicrobial properties and high UV-protecting activity.