Prevention, removal and inactivation of Escherichia coli and Staphylococcus aureus biofilms using selected monoterpenes of essential oils.

AIMS: The aim of this study was to investigate the antibiofilm potential of five essential oil (EO) components with cyclic (sabinene-SAB, carveol-C1, carvone-C2) and acyclic (citronellol-C3 and citronellal-C4) structures against Escherichia coli and Staphylococcus aureus. METHODS AND RESULTS: The selected EO components prevented biofilm set-up, with C3 and C4 causing remarkable effects. When applied against pre-established biofilms, they promoted high biomass removal and inactivation of biofilm cells. Moreover, no viable E. coli biofilm cells were detected after exposure to SAB at 5 × MIC and 10 × MIC, and a significant viability decrease was observed for both bacteria with the other EO components. SAB, C3 and C4 caused the most prominent effects apparently due to their octanol-water partition coefficient (Po/w), the number of rotatable bonds (n-ROTB) and the free hydroxyl groups. CONCLUSIONS: The overall results demonstrated that the selected EO components, particularly SAB, C3 and C4 are of interest as new lead molecules to both prevent biofilm set-up and to control pre-established biofilms of E. coli and S. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: The tested EO components exhibited prominent antibiofilm properties against E. coli and S. aureus providing a novel and effective alternative/complementary approach to counteract chronic infections and the transmission of diseases in clinical settings.

Disinfectant test against monoculture and mixed-culture biofilms composed of technological, spoilage and pathogenic bacteria: bactericidal effect of essential oil and hydrosol of Satureja thymbra and comparison with standard acid-base sanitizers.

AIMS: To assess the antimicrobial action of three natural-derived products (essential oil, decoction and hydrosol of Satureja thymbra) against biofilms, composed of useful, spoilage and pathogenic bacteria (formed as monoculture or/and mixed-culture), and to compare their efficiency with three standard acid and alkaline chemical disinfectants. METHODS AND RESULTS: Two acids (hydrochloric and lactic, pH 3), one alkali (sodium hydroxide, pH 11), the essential oil of S. thymbra (1% v/v) and the two by-products of the essential oil purification procedure (the decoction and the hydrosol fraction of essential oil, 100%), were tested against biofilms formed by five bacterial species, either as monospecies, or as mixed-culture of all species. The tested bacterial species were Staphylococcus simulans and Lactobacillus fermentum (useful technological bacteria), Pseudomonas putida (spoilage bacterium), Salmonella enterica and Listeria monocytogenes (pathogenic bacteria). Biofilms were left to be formed on stainless steel coupons for 5 days at 16 degrees C, before the application of disinfection treatments, for 60 and 180 min. The disinfection efficiency was evaluated by detaching the remaining viable biofilm cells and enumerating them by agar plating, as well as by automated conductance measurements (using Rapid Automated Bacterial Impedance Technique). Both these methods revealed that the essential oil and the hydrosol of S. thymbra exhibited a strong antimicrobial action against both monospecies and mixed-culture biofilms. Surprisingly, the efficiency of the other three acid-base disinfectants was not adequate, although a long antimicrobial treatment was applied (180 min). CONCLUSIONS: The essential oil of S. thymbra (1%), as well as its hydrosol fraction (100%), presents sufficient bactericidal effect on bacterial biofilms formed on stainless steel. SIGNIFICANCE AND IMPACT OF THE STUDY: Use of natural antimicrobial agents could provide alternative or supplemented ways for the disinfection of microbial-contaminated industrial surfaces.

Effect of temperature, pH, and water activity on biofilm formation by Salmonella enterica enteritidis PT4 on stainless steel surfaces as indicated by the bead vortexing method and conductance measurements.

An assay was developed in an effort to elucidate the effect of important environmental parameters (temperature, pH, and water activity [aw]) on Salmonella Enteritidis biofilm formation on stainless steel surfaces. To achieve this, a modified microbiological technique used for biofilm studying (the bead vortexing method) and a rapid method based on conductivity measurements were used. The ability of the microorganism to generate biofilm on the stainless surfaces was studied at three temperatures (5, 20, and 37 degrees C), four pH values (4.5, 5.5, 6.5, and 7.4), and four aw values (0.5, 1.5, 5.5, and 10.5% NaCl). Results obtained by the bead vortexing method show that maximum numbers of adherent bacteria per square centimeter (106 CFU/cm2) were attained in 6 days at 20 degrees C. Biofilm formation after 7 days of incubation at 20 degrees C was found to be independent of the pH value. In addition, the high concentration of sodium chloride (10.5% NaCl, aw = 0.94) clearly inhibited the adherence of cells to the coupons. Conductance measurements were used as a supplementary tool to measure indirectly the attachment and biofilm formation of bacterial cells on stainless steel surfaces via their metabolic activity (i.e., changes in the conductance of the growth medium due to microbial growth or metabolism). Results obtained by conductance measurements corresponded well to those of the bead vortexing method. Furthermore, we were able to detect cells that remained attached on the metal surfaces even after vortexing via their metabolic activity. The results, except for demonstrating environmental-dependent Salmonella Enteritidis biofilm formation, indicated that traditional vortexing with beads did not remove completely biofilm cells from stainless steel; hence, conductance measurements seem to provide a more sensitive test capable to detect down to one single viable organism.