Efficacy of plant essential oils against foodborne pathogens and spoilage bacteria associated with ready-to-eat vegetables: antimicrobial and sensory screening

The objectives of this study were to evaluate the antimicrobial activity of plant essential oils (EOs) against foodborne pathogens and key spoilage bacteria pertinent to ready-to-eat vegetables and to screen the selected EOs for sensory acceptability. The EOs basil, caraway, fennel, lemon balm, marjoram, nutmeg, oregano, parsley, rosemary, sage, and thyme were evaluated. The bacteria evaluated were Listeria spp., Staphylococcus aureus, Lactobacillus spp., Bacillus cereus, Salmonella, Enterobacter spp., Escherichia coli, and Pseudomonas spp. Quantitative antimicrobial analyses were performed using an absorbance-based microplate assay. Efficacy was compared using MIC, the half maximum inhibitory concentration, and the increase in lag phase. Generally, gram-positive bacteria were more sensitive to EOs than were gram-negative bacteria, and Listeria monocytogenes strains were among the most sensitive. Of the spoilage organisms, Pseudomonas spp. were the most resistant. Oregano and thyme EOs had the highest activity against all the tested bacteria. Marjoram and basil EOs had selectively high activity against B. cereus, Enterobacter aerogenes, E. coli, and Salmonella, and lemon balm and sage EOs had adequate activity against L. monocytogenes and S. aureus. Within bacterial species, EO efficacy was dependent on strain and in some cases the origin of the strain. On a carrot model product, basil, lemon balm, marjoram, oregano, and thyme EOs were deemed organoleptically acceptable, but only oregano and marjoram EOs were deemed acceptable for lettuce. Selected EOs may be useful as natural and safe additives for promoting the safety and quality of ready-to-eat vegetables.     

The antimicrobial efficacy of plant essential oil combinations and interactions with food ingredients

The objective of this study was to evaluate the efficacy of plant essential oils (EOs) in combination and to investigate the effect of food ingredients on their efficacy. The EOs assessed in combination included basil, lemon balm, marjoram, oregano, rosemary, sage and thyme. Combinations of EOs were initially screened against Bacillus cereus, Escherichia coli, Listeria monocytogenes and Pseudomonas aeruginosa using the spot-on-agar test. The influence of varying concentrations of EO combinations on efficacy was also monitored using E. coli. These preliminary studies showed promising results for oregano in combination with basil, thyme or marjoram. The checkerboard method was then used to quantify the efficacy of oregano, marjoram or thyme in combination with the remainder of selected EOs. Fractional inhibitory concentrations (FIC) were calculated and interpreted as synergy, addition, indifference or antagonism. All the oregano combinations showed additive efficacy against B. cereus, and oregano combined with marjoram, thyme or basil also had an additive effect against E. coli and P. aeruginosa. The mixtures of marjoram or thyme also displayed additive effects in combination with basil, rosemary or sage against L. monocytogenes. The effect of food ingredients and pH on the antimicrobial efficacy of oregano and thyme was assessed by monitoring the lag phase and the maximum specific growth rate of L. monocytogenes grown in model media. The model media included potato starch (0, 1, 5 or 10%), beef extract (1.5, 3, 6 or 12%), sunflower oil (0, 1, 5 or 10%) and TSB at pH levels of 4, 5, 6 or 7. The antimicrobial efficacy of EOs was found to be a function of ingredient manipulation. Starch and oils concentrations of 5% and 10% had a negative impact on the EO efficacy. On the contrary, the EOs were more effective at high concentrations of protein, and at pH 5, by comparison with pH 6 or 7. This study suggests that combinations of EOs could minimize application concentrations and consequently reduce any adverse sensory impact in food. However, their application for microbial control might be affected by food composition, therefore, careful selection of EOs appropriate to the sensory and compositional status of the food system is required. This work shows that EOs might be more effective against food-borne pathogens and spoilage bacteria when applied to ready to use foods containing a high protein level at acidic pH, as well as lower levels of fats or carbohydrates.     

Impact of plant essential oils on microbiological,organoleptic and quality markers of minimally processed vegetables

The objectives of this study were to evaluate the efficacy of plant essential oils (EOs) for control of the natural spoilage microflora on ready-to-eat (RTE) lettuce and carrots whilst also considering their impact on organoleptic properties. Initial decontamination effects achieved using EOs were comparable to that observed with chlorine and solution containing oregano recorded a significantly lower initial TVC level than the water treatment on carrots (p < 0.05). No significant differences were found between the EO treatments and chlorine considering gas composition, color, texture and water activity of samples. The sensory panel found EO treatments acceptable for carrots throughout storage, while lettuce washed with the EO solutions were rejected for overall appreciation by Day 7. Correlating microbial and sensory changes with volatile emissions identified 12 volatile quality markers. Oregano might be a suitable decontamination alternative to chlorine for RTE carrots, while the identification of volatile quality markers is a useful complement to sensory and microbiological assessments in the monitoring of organoleptic property changes and shelf-life of fresh vegetables.Industrial relevanceThere is industrial demand for natural alternatives to chlorine, which is commonly used for decontamination of fresh produce but which has limitations with respect to antimicrobial efficacy and possible formation of carcinogenic compounds in water. Plant essential oils have proven antimicrobial and other bioactive properties, however their usefulness in foods can be mitigated by their high sensory impact. This study examined the application of EOs for fresh produce decontamination addressing control of spoilage microflora and improving shelf-life characteristics whilst also considering the impact on organoleptic properties. The effectiveness of oregano as a decontamination treatment was comparable with that of chlorine. Carrot discs treated with the EO regimes were acceptable in terms of sensory quality and appreciation, therefore oregano could offer a natural alternative for the washing and preservation of fresh produce. Combining EOs with other natural preservatives might minimize doses and reduce the impact on organoleptic properties of fresh vegetables.     

Antimicrobial activity of whey protein isolate edible films with essential oils against food spoilers and foodborne pathogens

The use of antimicrobial edible film is proposed as a means of improving food safety and extending the shelf-life of food systems by controlling the release of antimicrobials on food surfaces. In this work we first selected and studied 8 different essential oils (EOs) from plants, namely, oregano, clove, tea tree, coriander, mastic thyme, laurel, rosemary, and sage as natural antimicrobials against 2 gram-positive bacteria (Listeria innocua and Staphylococcus aureus) and 2 gram-negative bacteria (Salmonella enteritidis and Pseudomona fragi) by using the agar disk diffusion method. EOs from oregano, clove, and tea tree produced the largest surfaces of inhibition against the growth of the 4 bacterial strains tested. Second and following the assessment of compatibility, stable antimicrobial edible films based on whey protein isolate (WPI) with increasing concentrations (0.5% to 9%) of the 8 EOs were developed and tested for antimicrobial activity against the same gram-positive and gram-negative bacteria. WPI-edible films incorporating oregano or clove EO were found to have the most intense inhibitory effect of microbial growth. The bacterial strain gram-negative P. fragi presented the less susceptibility to the effect of those films. Moreover, only the edible films based on these 2 EOs were active against all 4 studied microorganisms. On the other hand, the edible films incorporating tea tree, coriander, mastic thyme, laurel, rosemary, or sage EOs even at high concentrations (7% to 9%) did not cause any antimicrobial effect against the pathogens S. aureus or S. enteritidis. PRACTICAL APPLICATION: Potential applications of this technology can introduce direct benefits to the food industry by improving safety and microbial product quality. The results of this research have direct application in the food industry with potential applications in various foodstuffs, including meat and poultry products where the control of spoilage bacteria such as P. fragi throughout their chilled storage or the improvement of food safety by controlling pathogens such as S. enteritidis are topics of particular interest for the industry.     

Impact of Essential Oils on the Taste Acceptance of Tomato Juice,Vegetable Soup,or Poultry Burgers

Despite the vast body of available literature on the possibilities of essential oils (EOs) as food preservatives or functional ingredients, the sensory impact of their addition to foods has barely been approached. This work focuses on the hedonic taste acceptance of 3 food products (tomato juice, vegetable soup, and poultry burgers) when they are incorporated with potentially antimicrobial concentrations (20 to 200 μL/L) of 6 selected EOs (lemon, pennyroyal mint, thyme, and rosemary) and individual compounds (carvacrol, p‐cymene). Although addition of 20 μL/L of pennyroyal mint or lemon EO did not change the taste acceptance of tomato juice, higher concentrations of these compounds or any concentration of the other 4 compounds did. In vegetable soup, the tolerance limit for rosemary EO, thyme EO, carvacrol, or p‐cymene was 20 μL/L, while the addition of 200 μL/L of lemon EO was accepted. Tolerance limits in poultry burgers were established in 20 μL/L for carvacrol and thyme EOs, 100 μL/L for pennyroyal mint EO and p‐cymene, and 200 μL/L for lemon and rosemary EOs. Moreover, incorporation of pennyroyal mint EO to tomato juice or poultry burgers, and enrichment of vegetable soup with lemon EO, could contribute to the development of food products with an improved sensory appeal.     

Combined effect of natural essential oils, modified atmosphere packaging, and gamma radiation on the microbial growth on ground beef

Selected Chinese cinnamon, Spanish oregano, and mustard essential oils (EOs) were used in combination with irradiation to evaluate their ability to eliminate pathogenic bacteria and extend the shelf life of medium-fat-content ground beef (23% fat). Shelf life was defined as the time when the total bacterial count reached 10(7) CFU/g. The shelf life of ground beef was determined for 28 days at 4 degrees C after treatment with EOs. The concentrations of EOs were predetermined such that sensory properties of cooked meat were maintained: 0.025% Spanish oregano, 0.025% Chinese cinnamon, and 0.075% mustard. Ground beef samples containing EOs were then packaged under air or a modified atmosphere and irradiated at 1.5 kGy. Ground beef samples (10 g) were taken during the storage period for enumeration of total mesophilic aerobic bacteria, Escherichia coli, Salmonella, total coliforms, lactic acid bacteria, and Pseudomonas. Mustard EO was the most efficient for reducing the total mesophilic aerobic bacteria and eliminating pathogenic bacteria. Irradiation alone completely inhibited the growth of total mesophilic aerobic and pathogenic bacteria. The combination of irradiation and EOs was better for reducing lactic acid bacteria (mustard and cinnamon EOs) and Pseudomonas (oregano and mustard EOs). The best combined treatment for extending the shelf life of ground beef for up to 28 days was EO plus irradiation (1.5 kGy) and modified atmosphere packaging.     

In vitro antimicrobial effects and mechanism of action of selected plant essential oil combinations against four food-related microorganisms

The aim of this study was to evaluate the antimicrobial efficacy of selected plant essential oil (EO) combinations against four food-related microorganisms. Ten EOs were initially screened against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Saccharomyces cerevisiae using agar disk diffusion and broth dilution methods. The highest efficacy against all the tested strains was shown when testing the oregano EO. EOs of basil and bergamot were active against the Gram-positive bacteria (S. aureus and B. subtilis), while perilla EO strongly inhibited the growth of yeast (S. cerevisiae). The chemical components of selected EOs were also analyzed by GC/MS. Phenols and terpenes were the major antimicrobial compounds in oregano and basil EOs. The dominant active components of bergamot EO were alcohols, esters and terpenes. For perilla EO, the major active constituents were mainly ketones. The checkerboard method was then used to investigate the antimicrobial efficacy of EO combinations by means of the fractional inhibitory concentration index (FICI). Based on an overall consideration of antimicrobial activity, organoleptic impact and cost, four EO combinations were selected and their MIC values were listed as follows: oregano–basil (0.313–0.313 μl/ml) for E. coli, basil–bergamot (0.313–0.156 μl/ml) for S. aureus, oregano–bergamot (0.313–0.313 μl/ml) for B. subtilis and oregano–perilla (0.313–0.156 μl/ml) for S. cerevisiae. Furthermore, the mechanisms of the antimicrobial action of EO combinations to the tested organisms were studied by the electronic microscopy observations of the cells and the measurement of the release of cell constituents. The electron micrographs of damaged cells and the significant increase of the cell constituents' release demonstrated that all EO combinations affected the cell membrane integrity.     

A Weibull model to describe antimicrobial kinetics of oregano and lemongrass essential oils against Salmonella Enteritidis in ground beef during refrigerated storage

The antimicrobial effect of oregano (Origanum vulgare L.) and lemongrass (Cymbopogon citratus (DC.) Stapf.) essential oils (EOs) against Salmonella enterica serotype Enteritidis in in vitro experiments, and inoculated in ground bovine meat during refrigerated storage (4 ± 2 °C) for 6 days was evaluated. The Weibull model was tested to fit survival/inactivation bacterial curves (estimating of p and δ parameters). The minimum inhibitory concentration (MIC) value for both EOs on S. Enteritidis was 3.90 μl/ml. The EO concentrations applied in the ground beef were 3.90, 7.80 and 15.60 μl/g, based on MIC levels and possible activity reduction by food constituents. Both evaluated EOs in all tested levels, showed antimicrobial effects, with microbial populations reducing (p ≤ 0.05) along time storage. Evaluating fit-quality parameters (RSS and RSE) Weibull models are able to describe the inactivation curves of EOs against S. Enteritidis. The application of EOs in processed meats can be used to control pathogens during refrigerated shelf-life.     

Antimicrobial effectiveness of oregano and sage essential oils incorporated into whey protein films or cellulose‐based filter paper

BACKGROUND: In this study the antimicrobial effectiveness of oregano and sage essential oils (EOs) incorporated into two different matrices, whey protein isolate (WPI) and cellulose‐based filter paper, was analysed. RESULTS: Antimicrobial properties of WPI‐based films containing oregano and sage EOs were tested against Listeria innocua, Staphylococcus aureus and Salmonella enteritidis. Oregano EO showed antimicrobial activity against all three micro‐organisms. The highest inhibition zones were against L. innocua. However, sage EO did not show antimicrobial activity against any of the micro‐organisms. Antimicrobial activity was confirmed for both EOs using cellulose‐based filter paper as supporting matrix, although it was significantly more intense for oregano EO. Inhibition surfaces were significantly greater when compared with those of the WPI films. This finding is likely due to the higher porosity and diffusivity of the active compounds in the filter paper. CONCLUSION: The interactions between the EOs and the films have a critical effect on the diffusivity of the active compounds and therefore on the final antimicrobial activity. As a result, to obtain active edible films, it is necessary to find the equilibrium point between the nature and concentration of the active compounds in the EO and the formulation of the film. Copyright © 2010 Society of Chemical Industry     

Antimicrobial Activity of Thyme (Tymus vulgaris) and Oregano (Origanum vulgare) Essential Oils against Some Food-borne Microorganisms

The aim of this study was to investigate antibacterial effects of oregano and thyme essential oils (EOs) on some food-borne bacteria. GC-MS analysis of EOs was performed in order to determine their composition and phenols were predominant constituents. The investigation of the antibacterial effects of EOs was performed on Salmonella Enteritidis, Salmonella Thyphimurium, Staphylococcus aureus, methicillin resistant Staphylococcus aureus, Escherichia coli and Bacillus cereus, and MICs were determined by broth microdilution method. EOs exhibited antibacterial activity against all tested microorganisms.     

Caffeic acid derivatives in dried Lamiaceae and Echinacea purpurea products

The concentrations of caffeic acid derivatives within Lamiaceae and Echinacea (herb, spice, tea, and dietary supplement forms) readily available in the US marketplace (n = 72) were determined. After the first identification of chicoric acid in Ocimum basilicum (basil), the extent to which chicoric acid could be found within the family Lamiaceae was investigated. The dominant phenolic acid in all Lamiaceae samples was rosmarinic acid, which ranged from 2.04 mg/100 g (one of 12 oregano samples) to 622.28 mg/100 g (lemon balm). Of the herbs tested in this study (marjoram, oregano, peppermint, rosemary, sage, spearmint, and thyme from the family Lamiaceae), only basil and lemon balm (Melissa officinalis) contained chicoric acid. Basil samples (starting material and resulting end product) obtained from an industry cooperator, showed substantial phenolic deficiency as a result of processing (approximately 78% loss).     

Inactivation of Salmonella enterica serovar Typhimurium and Quality Maintenance of Cherry Tomatoes Treated with Gaseous Essential Oils

The antimicrobial activity of the essential oils (EOs) from cinnamon bark, oregano, mustard, and of their major components cinnamaldehyde, carvacrol, and allyl isothiocyanate (AIT) was evaluated as a gaseous treatment to reduce Salmonella enterica serovar Typhimurium in vitro and on tomatoes. In vitro tests showed that mustard EO and AIT had the greatest inhibition of Salmonella, followed by cinnamon EO and cinnamaldehyde, while oregano and carvacrol showed the least inhibition. Scanning electron microscopy images of S. Typhimurium on tomatoes suggest that the EOs and their major components damaged the bacteria, and the damage was more obvious after posttreatment storage at 10 °C for 4 and 7 d. Salmonella on inoculated tomatoes was reduced by more than 5 log colony forming units (CFU)/g by mustard EO and AIT, by 4.56 and 3.79 log CFU/g following cinnamon EO and cinnamaldehyde treatments, respectively, and 1.54 and 3.37 log CFU/g after oregano EO and carvacrol treatments, respectively. Mustard EO and AIT induced discoloration, softening, and loss of the vitamin C and lycopene during 21 d of storage at 10 °C, while treatment with cinnamon EO and cinnamaldehyde did not result in significant changes in tomato quality. Tomatoes treated with oregano EO had better quality than nontreated samples after storage. Therefore, treatment with cinnamon and oregano EO and their major components appeared to be feasible for inactivation of Salmonella on tomatoes and maintaining quality.     

Antibacterial Effects of Allspice, Garlic, and Oregano Essential Oils in Tomato Films Determined by Overlay and Vapor-Phase Methods

ABSTRACT:  Physical properties as well as antimicrobial activities against  Escherichia coli  O157:H7,  Salmonella enterica , and  Listeria monocytogenes  of allspice, garlic, and oregano essential oils (EOs) in tomato puree film-forming solutions (TPFFS) formulated into edible films at 0.5% to 3% (w/w) concentrations were investigated in this study. Antimicrobial activities were determined by 2 independent methods: overlay of the film on top of the bacteria and vapor-phase diffusion of the antimicrobial from the film to the bacteria. The results indicate that the antimicrobial activities against the 3 pathogens were in the following order: oregano oil > allspice oil > garlic oil.  Listeria monocytogenes  was less resistant to EO vapors, while  E. coli  O157:H7 was more resistant to EOs as determined by both overlay and vapor-phase diffusion tests. The presence of plant EO antimicrobials reduced the viscosity of TPFFS at the higher shear rates, but did not affect water vapor permeability of films. EOs increased elongation and darkened the color of films. The results of the present study show that the 3 plant-derived EOs can be used to prepare tomato-based antimicrobial edible films with good physical properties for food applications by both direct contact and indirectly by vapors emanating from the films.     

Anti yeast activities of some essential oils in growth medium, fruit juices and milk

The anti-yeast activities of four essential oils (EOs) from clary sage, juniper, lemon and marjoram against wild-type isolates of the food-related yeasts Geotrichum candidum, Pichia anomala, Saccharomyces cerevisiae and Schizosaccharomyces pombe in malt extract (ME) medium, apple juice and milk were investigated. Minimal inhibitory concentrations (MICs) for the EOs and their main components were determined and the checkerboard method was used to calculate fractional inhibitory concentration (FIC) indices for the combinations of EOs or components. The most sensitive yeast was S. pombe (MICs of 0.0625-0.125 μl/ml) while G. candidum proved to be the most insensitive (MICs of 0.5-2 μl/ml). In general, the lag phases were lengthened by increasing EO concentrations, while significant reduction of growth rates was obtained only at the highest EO concentrations. The anti-yeast effects of the EOs were good in the acidic pH range optimal for yeasts growth. Combinations of juniper and clary sage EOs resulted in additive effects in the case of S. cerevisiae and G. candidum, but all other combinations showed no interaction. The combination of α-pinene and limonene led to synergism, while the combination of α-pinene with linalool resulted in an additive effect. Cloudy apple juice protected the yeasts against the effect of lemon EO: the lag phases were shorter and the growth rates higher than in clear apple juice. Lemon EO decreased the growth rate of G. candidum in skimmed milk in a dose-independent manner. Our results show that by adding lemon EO to clear apple juice a new, harmonic taste can be achieved and open storage time could be prolonged.     

Antimicrobial activity of soy edible films incorporated with thyme and oregano essential oils on fresh ground beef patties

Antibacterial activity of soy protein edible films (SPEF) incorporated with 1, 2, 3, 4 and 5% oregano (OR) or thyme (TH) essential oils was evaluated against Escherichia coli, E. coli O157:H7, Staphylococcus aureus, Pseudomonas aeruginosa and Lactobacillus plantarum by the inhibition zone test. Effects of SPEF containing 5% OR and TH or a mixture of OR + TH (ORT) were also tested on fresh ground beef during refrigerated storage (at 4 °C). OR and TH incorporated SPEF exhibited similar antibacterial activity against all bacteria in inhibition zone test. While E. coli, E. coli O157:H7 and S. aureus were significantly inhibited by antimicrobial films, L. plantarum and P. aeruginosa appeared to be the more resistant bacteria. SPEF with OR, ORT, and TH did not have significant effects on total viable counts, lactic acid bacteria and Staphylococcus spp. when applied on ground beef patties whereas reductions (p < 0.05) in coliform and Pseudomonas spp. counts were observed.     

Antimicrobial activity of oregano oil on iceberg lettuce with different attachment conditions

In this study, the antimicrobial activity of oregano oil was investigated under different attachment conditions of Salmonella spp. to iceberg lettuce. Inoculated lettuce was either not dried or dried for 30 min, 60 min, or 120 min, under either static air or moving air. Washing iceberg lettuce with 500 ppm oregano oil for 1, 5, and 10 min reduced the population of Salmonella spp. by (respectively) 1.3, 1.65, and 2.28 log cfu/g following the most challenging inoculation conditions, an inoculum drying period of 2 h under moving air. Across all inoculation conditions, increasing the treatment time significantly increased the reductions in the populations of Salmonella spp. (P < 0.05). Browning and softening of the lettuce leaf surface was observed after 10 min of treatment with oregano oil. For each treatment time, attachment times and drying under static compared with moving air did not significantly affect the antimicrobial efficacy of the various oregano oil treatments (P > 0.05). The results obtained in this study suggest that oregano oil can effectively reduce populations of Salmonella attached to lettuce leaf surfaces. PRACTICAL APPLICATION: The use of essential oils as an antimicrobial treatment can help to ensure the safety of leafy green products. As used in this study, oregano oil effectively reduced Salmonella spp., even after the pathogen had dried onto the lettuce leaves. Treatments that incorporate oregano oil therefore hold promise as a biocide treatment for process and packaged lettuce.     

Synergistic antimicrobial activity of two binary combinations of marjoram,lavender, and wild thyme essential oils

The combination of essential oils (EOs) is a novel alternative to improve their preservative effects and to reduce their organoleptic impact in food. In this context, this work aims to investigate the antibacterial combined effect of two EOs combinations through the calculation of the fractional inhibitory concentration index. The combinations tested consists of Lavandula dentata/Origanum majorana EOs and Thymus serpyllum/Origanum majorana EOs. Their chemical compositions were identified by CG/MS analyses. The main compounds of O. majorana EO were (-) - terpinene-4-ol and trans-4-thujanol. Those of L. dentata EO were β-pinene and 1,8-cineole, and those of T. serpyllum EO were p-cymene and γ-terpinene. Regarding the outcomes, results highlighted partial synergistic and additive interactions. Two combinations of marjoram and thyme EOs had antibacterial activities against S. aureus. The first one corresponded to the quarter of the minimum inhibitory concentration of marjoram and half that of thyme. The second one was the mix of half and quarter of the minimum inhibitory concentration of respectively marjoram and thyme EOs. This last combination also showed an antibacterial effect against E. coli. The quarter and the half of their minimum inhibitory concentration of marjoram and lavender combination, respectively, gave a partial synergy against both strains. Henceforth, these findings could be largely exploited in food preservation through the use of minimal doses of these plant products without affecting the antibacterial and the organoleptic properties in foods.     

Modeling dependence of growth inhibition of Salmonella Typhimurium and Listeria monocytogenes by oregano or thyme essential oils on the chemical composition of minced pork

The aim of this study was to compare the antimicrobial effect of selected essential oils (EOs) at concentration of 1% in minced pork from five different meat cuts (loin, ham, shoulder, neck, and belly) and to assess the influence of the chemical parameters of meat on EO efficiency, determined by the decrease of pathogens in comparison to a control in samples after storage at 3°C/7 d in a vacuum. The inhibition was significant (p < .05) only in minced pork from meat cuts with a fat content lower than 5% (leg, loin and shoulder). Salmonella enterica serovar Typhimurium was more sensitive than Listeria monocytogenes and oregano was more effective than thyme (p < .001). The inhibition of S. Typhimurium depended mainly on the fat content (logarithmic dependence) and to a lesser degree on the pH, whereas pH was not found to be a significant predictor in the case of L. monocytogenes. The results of this study show that only lean minced meat (with a fat content max. 5%) is a suitable matrix for pathogen control by EOs.     

Effects of essential oils of oregano and nutmeg on growth and survival of Yersinia enterocolitica and Listeria monocytogenes in barbecued chicken

The in vitro effects of plant essential oils (EOs) against pathogenic bacteria are well known, yet few studies have addressed the effects of these compounds against pathogens associated with ready-to-cook foods. Experiments were conducted to determine the effectiveness of oregano and nutmeg EOs on the growth and survival of Yersinia enterocolitica and Listeria monocytogenes in broth culture and in Iranian barbecued chicken. Ready-to-cook Iranian barbecued chicken was prepared according to the common practice with 1, 2, and 3 microl/g of oregano and nutmeg EOs. The test and control (without EOs) samples were inoculated with Y. enterocolitica and L. monocytogenes to a final concentration of 6 to 7 log CFU/g and stored at 3, 8, and 20 degrees C. Microorganisms were counted just before and at 24, 48, and 72 h after storage based on growth on Yersinia selective agar supplemented with cefsulodine, igrasan, and novobiocin and on Listeria selective agar supplemented with nalidixic acid and acriflavin. In the broth culture system, the nutmeg EO had a greater effect on L. monocytogenes (MIC = 0.20 nicrol/ml) than did the oregano EO (MIC = 0.26 microl/ml). However, the oregano EO had a greater effect on Y. enterocolitica (MIC = 0.16 microl/ml) than did the nutmeg EO (MIC = 0.25 microl/ml). In ready-to-cook Iranian barbecued chicken, the log CFU per gram of both bacteria after up to 72 h of incubation was not decreased significantly by various combinations of oregano and nutmeg EOs (1, 2, and 3 microl/g) and storage temperatures (3, 8, and 20 degrees C) when compared with control samples (without EOs). Although examination of spices in culture media can yield accurate microbiological data, without complementary tests in foods these data are of limited value for assessing food safety.     

Preferential solubilization behaviours and stability of some phenolic-bearing essential oils formulated in different microemulsion systems

The solubilization behaviour of a number of essential oils (EOs) containing volatile phenolic constituents was investigated in five different micellar solutions. These oils include clove bud (Eugenia caryophyllata), thyme (Thymus serpyllum) and oregano (Thymus capitatus). Ternary and pseudo‐ternary phase diagrams were constructed to assess the ability for microemulsion formation and dilutability of each system using non‐ionic surfactants. Results showed that Tween 20 (T20) was more suitable to solubilize these oils compared with Tween 80 (T80). Clove EO was found to be easily microemulsifiable compared with the other EOs, whereas oregano showed the least tendency to form a microemulsion. Particle sizes measured at different dilution lines ranged between 5.9 and 16.9 nm. The chemical composition of each EO was revealed by gas chromatography and was correlated with the observed solubilization behaviour. The presence of solubilization enhancers like poly‐ols and short‐chain alcohols improved solubilization of all EOs; however, establishment of new dilution lines was controlled by EO type. Physical stability assessment showed that all microemulsions were stable against alternate freeze/thaw cycles which extended for 1 week. On the contrary, each system showed different temperature sensitivity in the thermal stress assessment. The results of this investigation can be useful in fabrication of thermodynamically stable aqueous system carrying aromatic and bioactive phenolics for different applications in personal hygiene, cosmetic, fragrance and pharmaceutical products.