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The combined inhibitory effect of essential oils (EOs) with meat-based marinades has not been fully studied. Therefore, the present study aimed to gauge the effect of a yogurt-based marinade when individually combined with three EOs, namely eugenol (EU), vanillin (VA), or β-resorcylic acid (BR) on camel meat cubes inoculated with Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7 during storage. Fresh camel meat cubes of 10 g were inoculated with bacteria and dipped in the mixture of marinade and EO. Overall, the study had six EO treatments (EU 0.5%, EU 1%, VA 0.5%, VA 1%, BR 0.5%, and BR 1%) and two controls (meat without marinade and marinated meat). Treated meat cubes were stored at 4°C or 10°C for 1, 4, and 7 d. Adding only marinade to the camel meat at 10°C decreased the pathogens by 0.8-2.4 log CFU/g. At 10°C, BR decreased L. monocytogenes, E. coli O157:H7, and Salmonella spp. by 2.0, 1.5, and 1.3 log CFU/g, while EU caused a decrease (p < 0.05) of 1.9, 1.2, and 0.9 log CFU/g, respectively. Similarly, VA caused a reduction in these microorganisms of 1.3, 1.1, and 1.0 log CFU/g, respectively (p < 0.05). The combination of marinade and EO resulted in a decrease of the pathogens ranging from 0.9-1.4 and 2.8-3.7 log CFU/g at 4 and 10°C, respectively. The antimicrobial efficacy of EO alone or when combined with marinade was higher at 10°C than at 4°C with all three pathogens at both 0.5% and 1%. Overall, EOs were found to enhance the microbial safety of camel meat. In addition, they are antimicrobials that occur naturally, require a minimum investment, and may prove to be a great asset for marinated camel meat producers., Competing Interests: Declaration of Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

The present study examined the influence of primary food components on the antifungal activity of the essential oil of Origanum vulgare, carvacrol, thymol, eugenol, and trans -cinnamaldehyde against Penicillium verrucosum and Aspergillus westerdijkiae . The MIC was determined in food model media enriched with proteins (1, 5, or 10%), carbohydrates (1, 4, or 6%), or oil (1, 5, or 10%). Proteins increased the antifungal activity of O. vulgare essential oil, carvacrol, thymol, and eugenol, whereas the effect of trans -cinnamaldehyde decreased with increasing protein content. The presence of carbohydrates diminished the inhibitory effect of the natural preservatives on A. westerdijkiae ; for P. verrucosum, their inhibitory effect increased with carbohydrates. Only the antifungal activity of trans -cinnamaldehyde did not depend on the carbohydrate content. The presence of oil had the strongest influence. At a concentration of 1% oil, the antifungal activity decreased significantly, and at 10% oil, almost no inhibition was observed. To investigate the effect of the antifungal agents on the morphology of the target molds, they were grown on malt extract agar containing carvacrol and trans -cinnamaldehyde and were examined by scanning electron microscopy. The hyphae, conidiophores, vesicles, and phialides were severely altered and deformed, and spore formation was clearly suppressed.

This study was conducted to investigate antibacterial properties of the colloidal silver nanoparticles (SNPs) and eugenol, alone and in combination, on Staphylococcus aureus and Salmonella Typhimurium and their interactions with food constituents (fat, protein, and carbohydrate). We examined antibacterial activities of SNPs and eugenol in Luria-Bertani (LB) broth and 1.5 and 3% fat ultrahigh-temperature (UHT) milk. MICs of eugenol and SNPs (particle size of 31.3 nm) were also investigated in the presence of sunflower oil, meat extract, and starch at concentrations of 2, 5, and 10% to examine the interactions between food constituents and antimicrobial agents. MICs and MBCs of eugenol and SNPs for both bacteria were at 2,500 and 25 μg/mL, respectively. Combinations of the two substances had additive and synergistic effects on Salmonella Typhimurium and S. aureus, respectively. Both compounds had bactericidal activity. In food matrices, results indicated that eugenol only in sunflower oil at 5 and 10% concentrations had significant antibacterial activity. A similar result was achieved for SNPs with 10% meat extract. In LB broth, eugenol at 2,500 and 5,000 μg/mL achieved 6-log reductions in the microbial population of both bacteria after 3 h, while SNPs achieved the same effect after 9 h. In UHT milk with 1.5% fat, eugenol at 5,000 μg/mL and SNPs at 25 μg/mL achieved 6-log reductions in bacterial populations after 24 h. Thus, the antimicrobial activity of both eugenol and SNPs depended on the medium in which the experiment was conducted, and the combination of both antimicrobial agents increased the antimicrobial effect.

The aim of the present study was to evaluate the quorum sensing (QS) inhibition potential of eugenol and eugenol nanoemulsion against QS-dependent virulence factor production and gene expression, as well as biofilm formation in Pseudomonas aeruginosa. In the current study, eugenol nanoemulsion at a sub-MIC of 0.2 mg/mL specifically inhibited about 50% of the QS-mediated violacein production in Chromobacterium violaceum, as well as the production of N-(3-oxododecanoyl)-l-homoserine lactone (3-oxo-C 12 -HSL) and C 4 -HSL N-acyl homoserine lactone signal molecules, pyocyanin, and swarming motility in P. aeruginosa. The inhibitive effect of eugenol and its nanoemulsion on the expression of the QS synthase genes was concentration dependent, displaying 65 and 52% expression level for lasI, respectively, and 61 and 45% expression level for rhlI, respectively, at a concentration of 0.2 mg/mL. In addition, the inhibitive effect of eugenol and its nanoemulsion on the expression of the rhlA gene responsible for the production of rhamnolipid was also concentration dependent, displaying 65 and 51% expression level for the rhlA gene, respectively, at a concentration of 0.2 mg/mL. Eugenol and its nanoemulsion also displayed 36 and 63% respective inhibition of biofilm formation by P. aeruginosa at the 0.2 mg/mL concentration. Therefore, the nanoemulsion could be used as a novel QS-based antibacterial and antibiofilm agent for the control of harmful bacteria.

Eugenol is a major phenolic component with diverse biological activities. However, it is difficult to formulate into an aqueous solution due to poor water solubility, and this limits its application. In the present study, eugenol nanoliposomes (EN) were prepared by combining the ethanol injection method with the dynamic high-pressure microfluidization method. Good physicochemical characterizations of EN were obtained. The successful encapsulation of eugenol in nanoliposomes was confirmed by Fourier transform infrared spectroscopy. A good storage stability of EN was confirmed by its low variation of average particle diameter and encapsulation efficiency after 8 weeks of storage. No oil drops were found in EN after 8 weeks of storage at 4°C and at room temperature, which suggested that the poor water solubility of eugenol was overcome by nanoliposome encapsulation. Compared with that of eugenol solution, a relatively good sustained release property was observed in EN. The antibacterial activity of EN against four common foodborne pathogenic bacteria (Staphylococcus aureus, Escherichia coli, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes) was evaluated in both Luria broth and milk medium.

Carvacrol and eugenol were encapsulated in micellar nonionic surfactant solutions to increase active component concentrations in the aqueous phase and used to treat two strains of Listeria monocytogenes (Scott A and 101) and two strains of Escherichia coli O157:H7 (4388 and 43895) grown as biofilms in a Centers for Disease Control and Prevention reactor. L. monocytogenes biofilms were grown in two different growth media, 1:20 TSB and Modified Welshimer's broth (MWB), while E. coli O157:H7 was grown in M9. In general, L. monocytogenes strains were more resistant to both micelle-encapsulated antimicrobials than E. coli O157:H7 strains. The two antimicrobials were equally effective against both strains of E. coli O157:H7, decreasing viable counts by 3.5 to 4.8 log CFU/cm(2) within 20 min. For both bacteria, most of the bactericidal activity took place in the first 10 min of antimicrobial exposure. Biofilm morphology and viability were assessed by the BacLight RedoxSensor CTC Vitality kit and confocal scanning laser microscopy, revealing an increasing number of dead cells when biofilms were treated with sufficiently high concentrations of carvacrol- or eugenol-loaded micelles. This study demonstrates the effectiveness of the application of surfactant-encapsulated essential oil components on two pathogen biofilm formers such as E. coli O157:H7 and L. monocytogenes grown on stainless steel coupons.

The antifungal activity of eugenol in a model system against aspergilli (Aspergillus niger, Aspergillus terreus, and Emericella nidulans), penicilli (Penicillium expansum, Penicillium glabrum, and Penicillium italicum), and fusaria (Fusarium oxysporum and Fusarium avenaceum) was investigated. Minimum detection time (time to attain a colony diameter of 1 cm) and the kinetic parameters were evaluated. The effectiveness of the active compound seemed to be strain or genus dependent; 100 mg/liter represented a critical value for P. expansum, P. glabrum, P. italicum, A. niger, and E. nidulans because a further increase of eugenol resulted in fungistatic activity. The radial growth of A. terreus and F. avenaceum was inhibited at 140 mg/liter, and growth of F. oxysporum was completely inhibited at 150 mg/liter.

This article reports on the investigation on the bioactivity of eugenol, limonene, and citrus extract against three bacteria (Lactobacillus plantarum, Lactobacillus brevis, and Bacillus coagulans) and three yeasts (Saccharomyces bayanus, Pichia membranifaciens, and Rhodotorula bacarum), representing the spoilage microflora of fruit juices. The experiments were performed with laboratory media by using a microdilution method. Data were fitted using the Gompertz equation, and the kinetic parameters were used to evaluate the MIC and the dose-dependent effect (at suboptimal doses for each essential oil). Citrus extract was the most effective essential oil, and the results suggested the following susceptibility hierarchy, from the most sensitive microorganism to the most resistant one (values in parentheses represent MICs): S. bayanus (2 ppm) > R. bacarum (3 ppm) > P. membranifaciens (5 ppm) > B. coagulans (cells, 20 ppm) > L. brevis (40 ppm) > L. plantarum (>40 ppm).

Cinnamon, clove, and lemongrass essential oils (EOs) and their active compounds cinnamaldehyde, eugenol, and citral, respectively, were investigated for their effectiveness as antimicrobial agents in an alginate-based edible coating (EC) on fresh-cut Fuji apples. This EC also contained malic acid, N-acetyl-L-cysteine, glutathione, and calcium lactate as quality stabilizing compounds. The EC applied on apple pieces effectively maintained the physicochemical characteristics of the apple pieces for more than 30 days, decreased the respiration rate, reduced the Escherichia coli O157:H7 population by about 1.23 log CFU/g at day 0, and extended the microbiological shelf life by at least 19 days. The addition of EOs at 0.7% (vol/vol) or their active compounds at 0.5% (vol/vol) into the EC increased its antimicrobial effect, reduced the E. coli O157:H7 population by more than 4 log CFU/g, and extended the microbiological shelf life by more than 30 days. However, those concentrations of EOs affected the physicochemical characteristics of fresh-cut apples and thus limited their shelf life from 7 to 21 days. Lemongrass and cinnamon EOs (0.7%), citral (0.5%), and cinnamaldehyde (0.5%) were the most effective compounds for extending microbiological shelf life, whereas lemongrass, cinnamon, and clove EOs at 0.3% (vol/vol) best maintained the physicochemical characteristics of the product. Apple pieces with EC at day 0 and with EC with or without lemongrass EO at 0.7% at day 15 were preferred by the panelists. ECs containing natural antimicrobials and quality stabilizing compounds may be useful for extending the shelf life of fresh-cut fruits.

The inhibitory effect of some plant oil aromatics against three strains of Arcobacter butzleri, two strains of Arcobacter cryaerophilus, and one strain of Arcobacter skirrowii was evaluated. When MICs were determined using the broth macrodilution method, cinnamaldehyde was most inhibitory followed by thymol, carvacrol, caffeic acid, tannic acid, and eugenol (P < 0.001). Sublethal concentrations of the three most potent plant oil aromatics also were examined. Overall, cinnamaldehyde was the most bacteriostatic against all arcobacters tested except A. butzleri when these strains were exposed to the MIC25 of this aromatic aldehyde. The bacteriostatic activities of thymol and carvacrol were concentration and species dependent.

The antimicrobial activity of eugenol microemulsions (eugenol encapsulated in surfactant micelles) in ultrahigh-temperature pasteurized milk containing different percentages of milk fat (0, 2, and 4%) was investigated. Antimicrobial microemulsions were prepared from a 5% (wt) aqueous surfactant solution (Surfynol 485W) with 0.5% (wt) eugenol. Two strains each of Listeria monocytogenes and Escherichia coli O157:H7 previously shown to be the least and most resistant to the microemulsion in microbiological media were used to inoculate sterile milk (10(4) CFU/ml). Samples were withdrawn and plated at 0, 1, 3, 6, 12, and 24 h for enumeration. Microemulsions completely prevented growth of L. monocytogenes for up to 48 h in skim milk and reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h. Similarly, in 2% fat milk, eugenol-Surfynol combinations reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h but only increased the lag phase of both strains of L. monocytogenes. In full-fat milk (4% fat), microemulsions inhibited growth of the least resistant strains of L. monocytogenes and E. coli but were ineffective against the two resistant strains. Unencapsulated eugenol was slightly more or as inhibitory as microemulsions against target pathogens. Results were attributed to diffusional mass transport of antimicrobials from microemulsions to the macroemulsion (milk). Results suggest that food composition, especially fat level, may affect the efficiency of targeting of foodborne pathogens with surfactant-encapsulated antimicrobials.

Growth inhibition of four strains of Escherichia coli O157:H7 (H1730, F4546, 932, and E0019) and Listeria monocytogenes (Scott A, 101, 108, and 310) by essential oil components (carvacrol and eugenol) solubilized in nonionic surfactant micelles (Surfynol 465 and 485W) was investigated. Concentrations of encapsulated essential oil components ranged from 0.02 to 1.25% depending on compound, surfactant type, and surfactant concentration (0.5 to 5%). Eugenol encapsulated in Surfynol 485W micelles was most efficient in inhibiting growth of the pathogens; 1% Surfynol 485W and 0.15% eugenol was sufficient to inhibit growth of all strains of E. coli O157:H7 and three of four strains of L. monocytogenes (Scott A, 310, and 108). The fourth strain, L. monocytogenes 101, was inhibited by 2.5% Surfynol and 0.225% eugenol. One percent Surfynol 485W in combination with 0.025% carvacrol was effective in inhibiting three of four strains of E. coli O157:H7. Strain H1730 was the most resistant strain, requiring 0.3% carvacrol and 5% surfactant for complete inhibition. Growth inhibition of L. monocytogenes by combinations of carvacrol and Surfynol 465 ranged between 0.15 and 0.35% and 1 and 3.75%, respectively. Generally, the antimicrobial activity of Surfynol 465 in combination with eugenol was higher than that for the combination with carvacrol. The potent activity was attributed to increased solubility of essential oil components in the aqueous phase due to the presence of surfactants and improved interactions of antimicrobials with microorganisms.

Growth inhibition of four strains of Escherichia coli O157:H7 (H1730, F4546, 932, and E0019) and Listeria monocytogenes (Scott A, 101, 108, and 310) by eugenol encapsulated in water soluble micellar nonionic surfactant solutions (Surfynol 485W) adjusted to pH 5, 6, and 7 and incubated at 10, 22, and 32 degrees C was determined. Concentrations of eugenol ranged from 0.2 to 0.9% at a surfactant concentration of 5%. Antimicrobial activity was assessed using a microbroth dilution assay. Eugenol encapsulated in surfactant micelles inhibited both microorganisms at pH 5, 6, and 7. At pH 5, some inhibition occurred in the absence of eugenol, i.e., by the surfactant itself (optical density at 24 h for L. monocytogenes = 0.07 and optical density at 24 h for E. coli O157:H7 = 0.09), but addition of >0.2% eugenol led to complete inhibition of both microorganisms. Inhibition of L. monocytogenes and E. coli O157:H7 decreased with increasing pH, that is, the minimum inhibitory concentration was 0.2, 0.5, and 0.5% of micellar encapsulated eugenol solutions at pH 5, 6, and 7, respectively. The encapsulated essential oil component in surfactant micelles was effective at all three temperatures tested (10, 22, and 32 degrees C), indicating that the activity of encapsulated eugenol was not affected by high or low (refrigeration) temperatures. Overall, strains of E. coli O157:H7 were more sensitive than strains of L. monocytogenes. Improved activity was attributed to increased solubility of eugenol in the aqueous phase due to the presence of surfactants and improved interactions of antimicrobials with microorganisms.

The application of combined preservative factors (hurdle technology) is very effective in controlling the growth of food spoilage and foodborne pathogenic bacteria. Antimicrobial activity of nisin alone and in combination with some natural organic compounds (carvacrol, cinnamic acid, eugenol, diacetyl, and thymol) on the growth of gram-positive bacteria Bacillus subtilis and Listeria innocua was-investigated. All the organic compounds tested exhibited antimicrobial activity against the microorganisms used; however, the MICs varied between 0.8 and 15.0 mM depending on the potency of the compound or the sensitivity of the target strain. Investigation of the interaction between the organic compounds and nisin against the test organisms revealed different patterns, varying from synergistic to antagonistic. Combinations of nisin with carvacrol, eugenol, or thymol resulted in synergistic action against both test organisms. Activity of nisin and cinnamic acid together was synergistic against L. innocua, but only additive against B. subtilis. In contrast, the combination of diacetyl and nisin resulted in an antagonistic effect against both test organisms. This study highlights the potential of the combination of these compounds with nisin to inhibit pathogen growth in food.

The use of multiple freeze (-20 degrees C)-thaw cycles in combination with isoeugenol and polysorbate 80 was investigated as a method for the reduction of numbers of Listeria monocytogenes cells in a bacteriological medium. Three freeze (1 h, -20 degrees C)-thaw cycles in the presence of isoeugenol at concentrations of 0, 100, and 300 ppm resulted in average L. monocytogenes reductions of 0.69, 2.65, and 3.3 log10 MPN (most probable number) per ml, respectively. Increasing the number of freeze-thaw cycles further decreased cell numbers, with reductions of nearly 5 log10 MPN/ml being obtained with six freeze-thaw cycles. Freeze-thaw cycles were effective in reducing cell numbers at isoeugenol concentrations down to 25 ppm. Rapid freezing rates with liquid nitrogen were found to be less effective in reducing numbers of L. monocytogenes cells. Two rapid freeze-thaw cycles in the presence of 100 ppm isoeugenol and polysorbate 80 resulted in a reduction of 1.45 log10 MPN/ml. Two freezing (-20 degrees C) cycles involving slow freezing and thawing rates with samples being held frozen for 6 h for each cycle resulted in reductions larger than those obtained with faster freezing rates. It was found that complete thawing in freeze-thaw cycles was not necessary to achieve bactericidal action. The application of multiple freeze-thaw cycles in combination with low concentrations of isoeugenol could effectively reduce numbers of L. monocytogenes cells in bacteriological media.

The effect of nine different oils was evaluated on the growth of Aspergillus parasiticus and Fusarium moniliforme. The experimental design to examine the inhibition of mycotoxins involved the incorporation of each of seven oils into broth and patty cultures. The fungal mycotoxin was identified by high-pressure liquid chromatography. Clove oil (eugenol) was the most inhibitory to the growth of A. parasiticus and F. moniliforme, followed by cinnamon (cinnamic aldehyde), oregano (thymol and carvacol) and mace oils (myristin). Neem and eucalyptus oil (cineole) did not affect fungal growth. The feasibility of implementing the results of this study to control mycotoxin toxicity was examined by costoring whole and ground cloves with mycotoxin-infected grain. Addition of both whole and ground cloves markedly reduced the aflatoxin contamination of the grain. These results clearly suggest that commonly occurring mycotoxigenic fungi can be controlled with clove oil (eugenol), thus spice oil successfully inhibited the growth of A. parasiticus and F. moniliforme, regulated the production of fumonisins. and prevented the formation of aflatoxins. The social implication of this finding is that rural communities can prevent the formation of fungal toxins in contaminated grain by simple measures.

Refrigerated ready-to-eat foods are becoming increasingly popular but are often vulnerable to contamination and subsequent growth by psychrotrophic foodborne pathogens. Consequently, there is a need for additional methods to assure the safety of these foods. Beef slices prepared from roasted whole sirloin tips were used in the study. Nine plant extracts were evaluated for ability to inhibit the growth of two psychrotrophic pathogens (Aeromonas hydrophila and Listeria monocytogenes) in refrigerated cooked beef. Results indicated that only eugenol (clove extract) and pimento extract significantly inhibited the growth of A. hydrophila and L. monocytogenes. However, L. monocytogenes was not as sensitive as was A. hydrophila to both treatments, especially to pimento extracts. These results suggest that plant extracts might be useful as an antimicrobial in cooked ready-to-eat meat.