Physical,antimicrobial and antibiofilm properties of Zataria multiflora Boiss essential oil nanoemulsion

It was evaluated physical, antibacterial and antibiofilm properties of Zataria multiflora Boiss essential oil (ZEO) and its nanoemulsion. Long‐term stability of nanoemulsion prepared by emulsion phase inversion was satisfying based on low narrow size distribution (polydispersity index ?0.2) and low droplet size (200 nm) over 21 days of storage. Nanoemulsion showed lower minimum inhibitory concentration (MIC) on Listeria monocytogenes (2500 µg mL^?1) than Salmonella Typhimurium (5000 µg mL^?1). Killing kinetics study revealed that nanoemulsion was more effective in inhibiting the growth of bacteria in milk than culture media. Both bacteriostatic and bactericidal effects were observed depending on the type of bacteria, nanoemulsion concentration and the time of exposure. Nanoemulsion at 4×MIC concentration reduced 64% and 75% of one‐day‐old biofilm of L. monocytogenes and S. Typhimurium, respectively. In conclusion, nanoemulsion revealed antimicrobial activity, but converting the ZEO to nanoemulsion did not improve its antibacterial activity; however, antibiofilm properties were enhanced.     

Zataria multiflora Boiss essential oil and sodium acetate: how they affect shelf life of vacuum‐packaged trout burgers

The preservative effects of Zataria multiflora Boiss essential oil (ZEO) at 0.025%, 0.05% and 0.1%, sodium acetate (SA) at 2% and their combination on the quality changes of vacuum‐packaged trout burgers during 21‐day refrigerated storage (4 ± 1°C) were investigated. Results showed that control and ZEO‐treated samples reached undesirable levels of rancidity (P < 0.05) at 15th day, whereas samples with the combined effect of SA and ZEOs, especially at higher concentrations, proved to be more stable in sensory, biochemical (PV, TBA, FFA and pH) and microbiological analyses (P < 0.05). Therefore, ZEO+SA‐treated samples showed good overall acceptability even until 21st day. Regarding the results obtained from ZEO‐treated samples, it becomes apparent that combined application of SA and ZEO extended the shelf life of fish burgers during cold storage to 21 days, 1.4‐fold longer than other treatments, which indicates the potential application of synergistic activity of these agents in vacuum‐packaged fishburgers.     

Formation and dispersal of biofilms in dairy substrates

The formation and dispersion of biofilms in the dairy industry is a problem because it increases cross‐contamination, affecting the shelf life of the products and their safety. The objective of this study was to evaluate the influence of different dairy substrates (cows’ milk and whey protein) on the formation and dispersion of Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus in two biofilm systems (mono‐species and multi‐species) on stainless steel at 25 °C. The dominant behaviour of E. faecalis occurred in most of the tests on mono‐species and multi‐species biofilms. A greater dispersion of biofilm cells was observed in skimmed milk.     

Evaluation of a novel antimicrobial (lauric arginate ester) substance against biofilm of Escherichia coli O157:H7, Listeria monocytogenes,and Salmonella spp.

The purpose of this study was to evaluate the activity of a novel antimicrobial substance lauric arginate ester (LAE) against selected foodborne pathogens (Escherichia coli O157:H7, Listeria monocytogenes and Salmonella spp.) in biofilm. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined and showed that LAE exhibits a strong antimicrobial activity. Biofilms were grown on abiotic stainless steel, rubber, MBEC biofilm device) and biotic (lettuce) surfaces. The efficacy of LAE (50, 100 and 200 ppm) at reducing the biofilm cells on these surfaces was examined by applying LAE for 2 h. Results revealed that LAE exhibited the reduction in biofilm bacteria up to 7 log CFU cm^?2, 3.5 log CFU cm^?2, 4.0 log CFU peg^?1 and 1.5 log CFU cm^?2 on stainless steel, rubber, MBEC and lettuce surfaces, respectively. Overall, these results suggest that LAE has been shown to be a potential alternative to control bacteria in biofilm mode in food industry.     

The Elimination of Listeria Monocytogenes Attached to Stainless Steel or Aluminum Using Multiple Hurdles

Ready‐to‐eat luncheon meats sliced in retail delis have been found to pose the greatest risk of foodborne illness from Listeria monocytogenes among all ready‐to‐eat foods. Slicers used in delis have many removable parts that are connected with seals and gaskets, with spaces, cracks and crevices that are difficult to clean adequately and may provide a niche for L. monocytogenes survival. Standard cleaning and sanitizing practices used by deli employees may not eliminate Listeria in these niches. Moist heat is known to be more effective against L. monocytogenes than dry heat at the same temperature and time. The study reported here investigated the effects of moist heat combined with quaternary ammonium compounds (5 or 10 ppm), chlorine (10 or 25 ppm) or peracetic acid (10 or 25 ppm) on inactivating L. monocytogenes attached to stainless steel or aluminum coupons cut from commercial deli meat slicer components. All sanitizers when used alone resulted in a 2‐ to 3‐log reduction of L. monocytogenes on stainless steel or aluminum surfaces, while moist heat alone resulted in a 3‐ to 4‐log reduction. When combined with heat the quaternary ammonium was used at 5 ppm, peracetic acid at 10 ppm and chlorine at 10 ppm. When the 2 lethal treatments were combined there was a 5‐ to7‐log reduction as compared to initial inoculation.     

Effectiveness of Zataria multiflora Boiss essential oil and grape seed extract impregnated chitosan film on ready-to-eat mortadella-type sausages during refrigerated storage

BACKGROUND: The effectiveness of chitosan films containing Zataria multiflora Boiss essential oil (ZEO) (5 and 10 g kg^?1) and grape seed extract (GSE) (10 g kg^?1) on lipid oxidation and microbial (lactic acid bacteria, aerobic mesophiles and inoculated Listeria monocytogenes) characteristics of mortadella sausage at 4 °C for 21 days was evaluated. The release of total phenolics (TPs) into sausage was also assessed. RESULTS: All films exhibited antibacterial activity against L. monocytogenes on agar culture media. Chitosan films containing ZEO were the most effective on the growth of bacteria. The growth of L. monocytogenes was significantly inhibited by ZEO‐GSE containing films especially during storage of the sausages for 6 days. Aerobic mesophiles and lactic acid bacteria were the most sensitive and resistant groups to films by 0.1–1.1 and 0.1–0.7 log cycles reduction, respectively. Sausages wrapped by 10 g kg^?1 GSE + 10 g kg^?1 ZEO films had the lowest degrees of lipid oxidation, which was 23% lower than the control. The TPs of ZEO films decreased to zero after 6 days, whereas TPs of GSE films followed a slight decrease during the storage. CONCLUSION: Antimicrobial/antioxidant chitosan film could be developed by incorporating GSE and ZEO for extending the shelf life of mortadella sausage. Copyright © 2011 Society of Chemical Industry     

Effect of partial replacement of potassium lactate and sodium diacetate by natural green tea and grape seed extracts and postpackaging thermal treatment on the growth of Listeria monocytogenes in hotdog model system

Low‐ (5%) and high‐fat (20%) chicken and turkey hotdogs were formulated in three groups: no antimicrobials (control), chemical preservatives (potassium lactate and sodium diacetate) alone and partial replacement of chemical preservatives by green tea (GTE) and grape seed extracts (GSE), surface inoculated (c. 10³ CFU g^?1) with Listeria monocytogenes, treated with or without heat treatment (65 °C for 104 s) to determine the growth of L. monocytogenes until spoilage (28 days). Maximum growth inhibitions (c. 2.0 CFU g^?1) were observed in the treatments having chemical preservatives and plant extracts regardless of the meat and fat type. Furthermore, plant extracts along with chemical preservatives demonstrated additional inhibitory effect on the growth of L. monocytogenes survivors in chicken hotdog samples followed by postpackaging heat treatment. Results demonstrated that natural GTE and GSE can partially replace the chemical preservatives and further enhance the antilisterial activities when combined with heat treatment.     

Investigation of some factors affecting the antibacterial activity of rosemary extracts in food models by a food microdilution method

The activity of rosemary phenolic extracts against Listeria monocytogenes and Escherichia coli was evaluated in 50% food models of meat, vegetable and dairy products in relation to some factors that can affect MIC (minimal inhibitory concentration) values (inoculum level, proportion of food, temperature) using a new food microdilution method. It was shown that the interactions of meat and milk components with plant extracts reduced the antibacterial effectiveness of rosemary extracts. MIC values for L. monocytogenes were lower than for E. coli in all tested conditions. A lower inoculation level caused a decrease in MIC values for E. coli but an increase in MIC values for L. monocytogenes in control media. In food models, MIC values were higher or equal to MIC values in control media regardless of bacterial type. We showed that the food microdilution method represents a simple, rapid, reproducible and inexpensive method for testing the antimicrobial efficiency of plant extracts in food systems.     

Aeromonas biofilm on stainless steel: efficiency of commonly used disinfectants

Aeromonas spp. are ubiquitous bacteria widely distributed among aquatic environments that have the ability to form biofilms. This aptitude allows them to persist in water distribution systems, contaminating drinking water, food processing surfaces and ultimately food. For this study, the biofilm‐forming ability of aeromonads was evaluated after 48‐h incubation on stainless steel discs at both 4 and 20 °C. Subsequently, disinfectants based on amphoteric surfactants and chlorine compounds were evaluated regarding the capacity to eradicate preformed biofilm and inhibit biofilm formation. Results obtained demonstrated that all strains under analysis were able to form biofilm at both room and refrigeration temperatures. The chlorine‐based disinfectant demonstrated to be efficient in removing preformed biofilm, but both were unsuccessful in preventing biofilm formation, highlighting the importance of adequate cleaning and disinfection procedures, with emphasis on food processing surfaces.     

Inhibitory effects of UV treatment and a combination of UV and dry heat against pathogens on stainless steel and polypropylene surfaces

Abstract: Pathogens that contaminate the surfaces of food utensils may contribute to the occurrence of foodborne disease outbreaks. We investigated the efficacy of UV treatment combined with dry heat (50 °C) for inhibiting 5 foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Pseudomonas aeruginosa, Listeria monocytogenes, and Staphylococcus aureus) on stainless steel and polypropylene surfaces in this study. We inoculated substrates with each of the 5 foodborne pathogens cultured on agar surface and then UV treatment alone or a combination of both UV and dry heat (50 °C) was applied for 30 min, 1 h, 2 h, and 3 h. The initial populations of the 5 pathogens before treatment were 8.02 to 9.18 and 8.73 to 9.16 log₁₀ CFU/coupon on the surfaces of stainless steel and polypropylene coupons, respectively. UV treatments for 3 h significantly inhibited S. Typhimurium, L. monocytogenes, and S. aureus on the stainless steel by 3.06, 2.18, and 2.70 log₁₀ CFU/coupon, and S. aureus on the polypropylene by 3.11 log₁₀ CFU/coupon, respectively. The inhibitory effects of the combined UV and dry heat treatment (50 °C) increased as treatment time increased, yielding significant reductions in all samples treated for 3 h, with the exception of S. aureus on polypropylene. The reduction level of E. coli O157:H7 treated for 3 h on the surface of stainless steel and polypropylene treated was approximately 6.00 log₁₀ CFU/coupon. These results indicate that combined UV and dry heat (50 °C) treatments may be effective for controlling microbial contamination on utensils and cooking equipment surfaces as well as in other related environments.     

Biofilm formation of Salmonella Typhimurium on stainless steel and acrylic surfaces as affected by temperature and pH level

The effect of temperature (28, 37 and 42 °C) and pH (6 and 7) on the biofilm formation capability of Salmonella Typhimurium on stainless steel and acrylic was investigated. The rate of biofilm formation increased with increasing temperature and pH, while the number of attached cells after 240 h decreased with increasing temperature and was not different between pH 6 and 7. The surface hydrophobicity of bacterial cells was not significantly (p > 0.05) different among tested conditions. Electron-donating/accepting properties changed with pH and temperature, although these changes did not correlate with the ability to form biofilms under respective conditions. Attachment of S. Typhimurium showed a preference for stainless steel compared to acrylic surfaces under all conditions tested. The results suggest that salmonellae were less adherent to acrylic than to stainless steel surfaces; thus, acrylic-type surfaces should be considered for use in the food industry over stainless steel where applicable. The rate of biofilm formation increased at higher temperatures and pH levels within the tested ranges. Hurdle technology using lower temperatures reduced pH may help delay biofilm formation on food contact surfaces contaminated with S. Typhimurium.     

Antilisterial effect of two bioprotective cultures in a model system of Iberian chorizo fermentation

This study reports the different effects of two bioprotective cultures on the growth of different Listeria monocytogenes strains by a rapid assay simulating the first stage of Iberian chorizo fermentation. Ground pork with or without protective cultures was inoculated with L. monocytogenes and incubated under two different conditions simulating the traditional, slow fermentation temperature (7 °C, 1 day) and a high, fast fermentation temperature (20 °C, 1 day), followed in both cases by storage at 7 °C for 13 days. Both bioprotective cultures reduced the growth of L. monocytogenes by at least 2 log CFU g^?1 at the end of both incubation periods compared with a noninoculated culture control lot. The best results were obtained with the strain Lactobacillus sakei CTC494, which exerted a bactericidal effect on L. monocytogenes under both conditions assayed, achieving a 5.4‐log reduction after 14 days compared with the control when the initial temperature was 20 °C.     

Detection of Salmonella spp., Yersinia enterocolitica,Listeria monocytogenes and Campylobacter spp. by real‐time multiplex PCR using amplicon DNA melting analysis and probe‐based assay

Syto9 and probe‐based multiplex real‐time PCR assays for simultaneous detection of a group of foodborne pathogens (named SYLC group), targeting Salmonella spp. (invA gene), Yersinia enterocolitica (ystA gene), Listeria monocytogenes (hly gene) and Campylobacter spp. (rrna gene), have been developed. The Syto9 assay generates amplicon DNA melting curve with four peaks of 86.5 ± 0.5, 84 ± 0.5, 81.5 ± 0.5 and 90.5 ± 0.5 °C corresponding Salmonella spp., Y. enterocolitica, L. monocytogenes and Campylobacter spp. targets, respectively. The sensitivities of the Syto9 and TaqMan assays in artificially inoculated chicken wing rinses were in a range of 3.2 × 10² to 3.1 × 10⁴ and 9.8 × 10² to 1.9 × 10⁴ colony‐forming units per millilitre, respectively, depending on the pathogen. All tested target strains (n = 100) were correctly detected by the both assays, whereas nontarget strains (n = 100) demonstrated no cross‐reactivity representing 100% specificity. The assays are suitable for application in qualitative and quantitative detection of SYLC group pathogens in food matrices.     

The prevention and removal of biofilm formation of Staphylococcus aureus strains isolated from raw milk samples by citric acid treatments

In this study, the antibiofilm activity of citric acid treatment on Staphylococcus aureus strains isolated from raw milk samples was evaluated. For this purpose, the prevention and removal of biofilm formation of S. aureus strains by citric acid treatments (2% and 10%) for 20 min were investigated for comparison with peracetic acid treatment (0.3%) on both microtitration plate and stainless steel coupons. The results indicated that the prevention and removal of biofilm formation and the numbers of prevented or removed S. aureus strains using citric acid treatments were observed to be higher than those using peracetic acid treatment on both surfaces. The prevention and removal of biofilm formation were substantially higher when the concentration of citric acid treatment increased from 2% to 10% and the stainless coupons were used. The results show that citric acid can be used as an alternative disinfectant in controlling biofilm formation in the dairy industry.     

The prevention of fish spoilage by high antioxidant Australian culinary plants: Shewanella putrefaciens growth inhibition

Shewanella putrefaciens is a marine bacterium and a major microbial cause of spoilage in low temperature stored seafood. A survey of fruits and culinary herbs was undertaken on Australian plants with high antioxidant capacities. Twenty‐eight extracts from thirteen plant species were investigated for the ability to inhibit S. putrefaciens growth. Of these, eight extracts (28.6%) substantially inhibited S. putrefaciens growth. The muntries (Kunzea pomifera), lemon aspen (Acronychia acidula) and desert lime (Citrus glauca) extracts were efficient anti‐S. putrefaciens agents, with MIC values ≤3000 μg mL^?1. Of these, the muntries methanolic extract was the most potent growth inhibitor (MIC = 2240 μg mL^?1). The aqueous desert lime extract was also an effective growth inhibitor (MIC of 3857 μg mL^?1), whilst the methanolic bush tomato (Solanum aviculare), aqueous muntries and Davidson's plum (Davidsonia pruriens) extracts displayed moderate S. putrefaciens growth inhibition. All extracts were nontoxic in the Artemia fransiscana bioassay, with LC50 values (>1000 μg mL^?1). Nontargeted HPLC‐QTOF mass spectroscopy (with screening against three compound databases) putatively identified twenty compounds that were present in both inhibitory muntries extracts. The low toxicity of these extracts and their inhibitory bioactivity against S. putrefaciens indicates their potential as natural fish and seafood preservatives.     

Synergistic effect between Helichrysum italicum essential oil and cold nitrogen plasma against Staphylococcus aureus biofilms on different food‐contact surfaces

Staphylococcus aureus is the most common pathogen in human, and the most diseases produced by S. aureus are associated with biofilms. Helichrysum italicum essential oil (EO), as a natural and safe spice, was employed to disperse S. aureus biofilm by cold nitrogen plasma (CNP) assist. After S. aureus biofilm formation on food container surfaces, they were exposed to CNP and then treated with Helichrysum italicumEO for biofilm dispersion. The population of S. aureus biofilm was approximately reduced by 2 logs after individual treatment of 0.5 mg mL^?1 Helichrysum italicumEO or 400 W CNP. Interestingly, the combined treatment of Helichrysum italicumEO (0.5 mg mL^?1, 40 min) and CNP (400 W, 1 min) reduced the S. aureus viable count in biofilm below 2 logs CFU per cm² after 1‐day storage. Therefore, the synergetic treatment holds great promise to improve the current treatment systems of bacterial contamination on different food‐contact surfaces.     

Assessment of bacterial biofilm on stainless steel by hyperspectral fluorescence imaging

Hyperspectral fluorescence imaging techniques were investigated for detection of two genera of microbial biofilms on stainless steel material which is commonly used to manufacture food processing equipment. Stainless steel coupons were deposited in nonpathogenic E. coli O157:H7 and Salmonella cultures, prepared using M9 minimal medium with casamino acids (M9C), for 6 days at 37 °C. Hyperspectral fluorescence emission images of the biofilm formations on the stainless coupons were acquired from 416 to 700 nm with the use of ultraviolet-A (320–400 nm) excitation. In general, emission peaks for both bacteria were observed in the blue region at approximately 480 nm and thus provided the highest contrast between the biofilms and background stainless steel coupons. A simple thresholding of the 480 nm image showed significantly larger biofilm regions for E. coli O157:H7 than for Salmonella. Viable cell counts suggested that Salmonella formed significantly higher density biofilm regions than E. coli O157:H7 in M9C medium. On the basis of principal component analysis (PCA) of the hyperspectral fluorescence images, the second principal component image exhibited the most distinguishable morphological differences for the concentrated biofilm formations between E. coli and Salmonella. E. coli formed granular aggregates of biofilms above the medium on stainless steel while Salmonella formed dense biofilm in the medium-air interface region (pellicle). This investigation demonstrated the feasibility of implementing fluorescence imaging techniques to rapidly screen large surface areas of food processing equipment for bacterial contamination. Company and product names are used for clarity and do not imply any endorsement by USDA to the exclusion of other comparable products.     

Modelling the cross‐contamination of Listeria monocytogenes in pork during bowl chopping

This study has developed a predictive model for the cross‐contamination of pork by Listeria monocytogenes during bowl chopping. The transfer rates of L. monocytogenes were measured in sixteen chopping scenarios based on practical work. Meanwhile, contaminated bowl chopper was cleaned with either a dry rag (DR), warm water (WW) or 70% ethanol + water (EW), respectively. It was showed that significant differences (P < 0.05) were observed among the three cleaning methods on the reduction of L. monocytogenes, the greatest log reduction being achieved by EW. Moreover, the model introduced by a previous study, predicting cross‐contamination of L. monocytogenes during meat slicing, was improved and validated in this study. Verification results showed that the improved model was acceptable for predicting L. monocytogenes cross‐contamination during pork chopping with coefficients of determination (R² > 0.82), accuracy factors (A_f < 1.44), bias factors (B_f < 1.42), and root mean square error (RMSE < 0.99). Furthermore, the modified model might provide an effective tool for assessing the risk of the cross‐contamination of meat products.     

Effect of carvacrol and thymol on Salmonella spp. biofilms on polypropylene

The present study evaluated the effects of carvacrol and thymol against Salmonella spp. biofilm on polypropylene. The efficacy of the compounds was assessed by quantifying Salmonella spp. cells during and after biofilm formation on polypropylene and performing scanning electron microscopy. During biofilm formation, carvacrol and thymol, at subinhibitory concentrations, reduced bacterial counts about 1–2 log, while established Salmonella spp. biofilms were reduced about 1–5 log by carvacrol and thymol, at MIC or 2× MIC. The greatest reduction in carvacrol‐treated biofilms, about 5 log, was observed with 156 and 312 μg mL^?1 (MIC and 2× MIC) in established Salmonella Typhimurium ATCC 14028 biofilms. Thymol showed the greatest reduction, about 4 log, at 624 μg mL^?1 (2× MIC) against mature Salmonella Enteritidis biofilm. Carvacrol and thymol reduced the number of Salmonella spp. cells on polypropylene, suggesting their potential for the control of Salmonella spp. biofilms.     

Fate of biofilm cells of Cronobacter sakazakii under modified atmosphere conditions

Survival of biofilm cells of Cronobacter sakazakii formed on stainless steel and polyvinyl chloride (PVC) on exposure to different atmosphere conditions was studied. Biofilms were formed on stainless steel and PVC coupons by using three strains of C. sakazakii. Six day old biofilms on stainless steel and PVC coupons were stored under N₂ gas, CO₂ gas, and air for up to 20 days. N₂ and CO₂ gases resulted in significant (p < 0.05) further reductions of 1.79 and 2.47 log CFU/cm² after 20 days of storage, respectively, compared to air storage. N₂ and CO₂ gases led to less reduction of biofilm cells on PVC compared to those on stainless steel. N₂ and CO₂ gases resulted in significant (p < 0.05) further reductions of 0.98 and 1.20 log CFU/cm² after 20 days of storage, respectively, compared to air storage.