ANTIFUNGAL ACTIVITIES OF THYME, CLOVE AND OREGANO ESSENTIAL OILS

The antifungal potential of essential oils of oregano (Origanum vulgare), thyme (Thymus vulgaris) and clove (Syzygium aromaticum) was determined. To establish this antifungal potential, two molds related to food spoilage, Aspergillus niger and Aspergillus flavus, were selected. The agar dilution method was employed for the determination of antifungal activities. The three essential oils analyzed presented inhibitory effects on both molds tested. Oregano essential oil showed the highest inhibition of mold growth, followed by clove and thyme. Aspergillus flavus was more sensitive to thyme essential oil than A. niger. Clove essential oil was a stronger inhibitor against A. niger than against A. flavus.     

Influence of Some Spice Essential Oils on Aspergillus Parasiticus Growth and Production of Aflatoxins in a Synthetic Medium

Gas-liquid chromatography was used to determine the essential oil compositions of thyme, cumin, clove, caraway, rosemary, and sage. The basic components of these oils were thymol, cumin aldehyde, eugenol, carvonc, borneol and thujonc, respectively. The antifungal potential of the oils against Aspergillus parasiticus were investigated. The essential oils caused complete inhibition of both mycelial growth and aflatoxin production. The effectiveness followed the sequence: thyme > cumin > clove > caraway > rosemary > sage. The major components of the essential oils produced an inhibitory effect at minimum inhibitory concentrations equal to those obtained with the oils.     

Effects of Tropical Citrus Essential Oils on Growth, Aflatoxin Production, and Ultrastructure Alterations of Aspergillus flavus and Aspergillus parasiticus

Ethyl acetate extracts and hydrodistillated essential oils from five cultivars of tropical citrus epicarps were evaluated for their inhibitory activities against Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus, Aspergillus parasiticus, and Penicillium sp. using disk diffusion and broth microdilution assays. Essential oils prepared from kaffir lime (Citrus hystrix DC) and acid lime (Citrus aurantifolia Swingle) epicarps exhibited stronger antifungal activity to all fungi than their ethyl acetate extracts with minimum inhibitory concentration and minimum fungicidal concentration values of 0.56 and 1.13 mg/ml (dry matter), respectively, against aflatoxin-producing A. flavus and A. parasiticus. The dominant components of the essential oil from kaffir lime were limonene, citronellol, linalool, o-cymene, and camphene, whereas limonene and p-cymene were major components of acid lime essential oil. Pure limonene, citronellal, and citronellol were five to six times less fungicidal than the natural essential oils, indicating the synergistic activity of many active compounds present in the oils. Kaffir and acid lime essential oils significantly reduced aflatoxin production of A. flavus and A. parasiticus, particularly lime essential oil, which completely inhibited growth and aflatoxin production of A. flavus at the concentration of 2.25 mg/ml. Target cell damage caused by acid lime essential oil was investigated under transmission electron microscopy. Destructive alterations of plasma and nucleus membrane, loss of cytoplasm, vacuole fusion, and detachment of fibrillar layer were clearly exhibited in essential-oil-treated cells.     

Lactobacillus plantarum 29 Inhibits Penicillium spp. Involved in the Spoilage of Black Truffles (Tuber aestivum)

The effect of an antifungal culture of Lactobacillus plantarum to be used in the storage at refrigeration temperature of fresh black truffles was examined. The strain was selected among 29 lactobacilli isolated from foods and evaluated for their viability and acidification activity at 4 °C, as well as for their inhibitory activity against 11 Penicillium strains isolated from truffles stored at refrigeration temperature. Lb. plantarum 29 showed the ability to hold not only the growth of Penicillium isolated from truffles, but also that of P. digitatum DSM 2750, a green mold involved in the spoilage of truffles. The antifungal activity was observed in vitro and in situ, and the sensory characteristics of truffles were preserved during the cold storage.     

Prolonging Storage Time of Baby Ginger by Using a Sand‐Based Storage Medium and Essential Oil Treatment

Wilt and rot occur readily during storage of baby ginger because of its tender skin and high moisture content (MC). A storage medium, which consisted of sand, 20% water, and 3.75% super absorbent polymers delayed weight loss and loss of firmness at 12 °C and 90% relative humidity. Microorganisms were isolated and purified from decayed rhizomes; among these, 3 fungi were identified as pathogens. The results of 18S rDNA sequence analysis showed that these fungi belonged to Penicillium, Fusarium, and Mortierella genera. The use of essential oil for controlling these pathogens was then investigated in vitro. Essential oils extracted from Cinnamomum zeylanicum (cinnamon) and Thymus vulgaris (thyme) completely inhibited the growth of all of the above pathogens at a concentration of 2000 ppm. Cinnamon oil showed higher antifungal activity in the drug sensitivity test with minimal fungicidal concentration (<500 ppm against all pathogens). In the in vivo test, cinnamon fumigation at a concentration of 500 ppm reduced infection rates of Penicillium, Fusarium, and Mortierella by 50.3%, 54.3%, and 60.7%, respectively. We recommended cinnamon oil fumigation combined with medium storage at 12 °C as an integrated approach to baby ginger storage.     

The effects of thyme (Thymus vulgaris) and rosemary (Rosmarinus officinalis) essential oils on Brochothrix thermosphacta and on the shelf life of beef packaged in high-oxygen modified atmosphere

The objective of the study was to determine the Minimal Inhibitory Concentration (MIC) of thyme (29.4% thymol, 21.6% p-cymene) and rosemary essential oils (27.6% 1,8-cineole, 13.5% limonene, 13.0% β-pinene) against Brochothrix thermosphacta and to establish the feasibility of their use as components of modified atmosphere during beef refrigerated storage. The minimum inhibitory concentration (MIC) of thyme oil against B. thermosphacta is 0.05% and that of rosemary oil 0.5%. The MIC values are independent on strain and temperature of growth, however the bactericidal effects are strain dependent. The addition of any of oil at a concentration equal to 2MIC to the modified atmosphere (80% O₂/20% CO₂) does not significantly influence the microbial quality of meat. At the same time, such a concentration of the essential oils was considerably detrimental to the organoleptic factors.     

Application of plant essential oils in prevention of fungal growth on Para rubber wood

Antifungal activities of essential oils (cinnamon, clove, eucalyptus, peppermint and lemongrass oils) against moulds (Penicillium sp., Aspergillus niger and Aspergillus versicolor) isolated from rubber wood surfaces were examined. Clove oil possessed consistent antifungal activity with the minimal inhibitory concentration (MIC) of 5 μl ml^?1 against all these fungi, while cinnamon oil had MICs from 2.5 to 10 μl ml^?1. However, only dip treatment with cinnamon oil inhibited the growth of A. niger on rubber wood for at least 25 weeks.     

Antifungal activity of 2 lactic acid bacteria of the Weissella genus isolated from food

Abstract: In the present study, a total of 116 lactic acid bacteria (LAB) strains isolated from Mill flour and fermented cassava were screened for their antifungal activity. Three strains among 116 were selected for their strongest inhibitory activity against food molds. These 3 strains were Lactobacillus plantarum VE56, Weissella cibaria FMF4B16, and W. paramesenteroides LC11. The compounds responsible for the antifungal activity were investigated. The strains displayed an inhibitory activity against targeted molds at acidic pH. However, the influence of organic acids was rejected according to the calculated minimal inhibitory concentration (MIC). Antifungal compounds were investigated in the cell‐free supernatants and phenyllactic acid (PLA) was detected in different amounts with a maximal concentration for Lb. plantarum VE56 (0.56 mM). Hydroxy fatty acid, such as 2‐hydroxy‐4‐methylpentanoic acid, was also produced and involved in the inhibitory activity of Lb. plantarum VE56 and W. paramesenteroides LC11. Antifungal LAB are known to produce PLA and 3‐hydroxy fatty acids and other organic acids with antifungal activity. This short communication focuses on antifungal activity from Weissella genus. The antifungal activity was attributed to antifungal compounds identified such as PLA, 2‐hydroxy‐4‐methylpentanoic acid, and other organic acids. Nevertheless, the concentration produced in the cell‐free supernatant was too low to compare to their MIC, suggesting that the inhibitory activity was caused by a synergy of these different compounds. Practical Application: Antifungal LAB are interesting to prevent food spoilage in fermented food and prolong their shelf life. In this way, chemical preservatives could be avoided and replaced by natural preservatives.     

Production and Characterization of an Antifungal Compound (3-Phenyllactic Acid) Produced by <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> Strain

The Lactobacillus plantarum strain was isolated from grass silage that produces a broad spectrum of antifungal compound, active against food and feed-borne filamentous fungi in agar plate assay. Aspergillus fumigatus and Rhizopus stolonifer were the most sensitive among molds. No inhibitory activity could be detected against mold Penicillium roqueforti. Enhanced antifungal activity was observed at 30 °C in pH 6.5. Minimum inhibitory concentration values against fungal cultures were ranged from 6.5 to 12.0 mg/ml for commercial 3-phenyllactic acid. The production of antifungal compound phenyllactic acid (PLA), lactic acid, and acetic acid by L. plantarum strain was also investigated. Structure characterization of the antifungal compound was carried out by nuclear magnetic resonance spectroscopy, infrared spectroscopy, and gas chromatography. The produced compound (PLA) acted as a fungistatic and delayed the growth of a variety of fungal contaminants.     

Inhibitory effect of cinnamon essential oils on selected cheese-contaminating fungi (Penicillium spp.) during the cheese-ripening process

The antifungal effects of essential oils (EOs) on cultures of the known cheese fungal contaminants Penicillium spp. were evaluated. Cinnamon leaf and bark EOs were the most effective among 8 EOs tested. The main components of both cinnamon EOs were eugenol, cinnamaldehyde, and linalool. Both inhibited growth of Penicillium spp. at a concentration of 10% v/v (2,000 ppm/mm²) on a lawn cell plate. When tested using 3 commercially available cheese starters, cinnamon EOs showed no effect against the FD-DVS ABT-5 cheese starter. However, growth of lactobacilli was inhibited in the presence of ≥10% (v/v) of leaf and bark EOs for the KAZU 1 starter, and ≥5% (v/v) for the FD-DVS FLORA-DANICA starter. A concentration of 4,000 ppm/mm² of cinnamon EOs completely inhibited growth of the Penicillium spp. that naturally contaminates the surface of Appenzeller cheese.     

Antifungal Effect of Allium tuberosum,Cinnamomum cassia,and Pogostemon cablin Essential Oils and Their Components Against Population of Aspergillus Species

Antifungal activity of Allium tuberosum (AT), Cinnamomum cassia (CC), and Pogostemon cablin (Patchouli, P) essential oils against Aspergillus flavus strains 3.2758 and 3.4408 and Aspergillus oryzae was tested at 2 water activity levels (a_w: 0.95 and 0.98). Main components of tested essential oils were: allyl trisulfide 40.05% (AT), cinnamaldehyde 87.23% (CC), and patchouli alcohol 44.52% (P). The minimal inhibitory concentration of the plant essential oils against A. flavus strains 3.2758 and 3.4408 and A. oryzae was 250 ppm (A. tuberosum and C. cassia), whereas Patchouli essential oil inhibited fungi at concentration > 1500 ppm. The essential oils exhibited suppression effect on colony growth at all concentrations (100, 175, and 250 ppm for A. tuberosum; 25, 50, and 75 for C. cassia; 100, 250, and 500 for P. cablin essential oil). Results of the study represent a solution for possible application of essential oil of C. cassia in different food systems due to its strong inhibitory effect against tested Aspergillus species. In real food system (table grapes), C. cassia essential oil exhibited stronger antifungal activity compared to cinnamaldehyde.     

Assessment of Thymus vulgaris L. essential oil as a safe botanical preservative against post harvest fungal infestation of food commodities

A total of 14 odoriferous angiospermic essential oils were tested against the toxigenic strain of Aspergillus flavus. The essential oil of Thymus vulgaris L. showed highest antifungal efficacy. The thyme oil absolutely inhibited the mycelial growth of A. flavus at 0.7 μl ml^− 1 and exhibited a broad fungitoxic spectrum against eight different food contaminating fungi viz. Fusarium oxysporum, Cladosporium herbarum, Curvularia lunata, Aspergillus terreus, Aspergillus niger, Aspergillus fumigatus, Alternaria alternata and Botryodiploidia theobromae. The oil also showed significant antiaflatoxigenic efficacy as it completely arrested the aflatoxin B₁ production at 0.6 μl ml^− 1. Thyme oil as fungitoxicant was also found superior over most of the prevalent synthetic fungicides. The LC₅₀ of thyme oil against mice was recorded as 7142.85 μl kg^− 1 body weight indicating its non-mammalian toxicity and strengthening its safe exploitation as preservative for stored food commodities. The findings recommend the thyme oil as potential botanical preservative in eco-friendly control of biodeterioration of food commodities during storage.

Industrial relevance

The thyme essential oil may be recommended for large scale application as a plant based preservative for stored food items because of its strong antifungal as well as antiaflatoxigenic efficacy. Because of broad antimicrobial spectrum, more efficacy over prevalent synthetic preservatives as well as non-mammalian toxicity, the thyme essential oil may be formulated as a safe and economical plant based preservative against post harvest fungal infestation and aflatoxin contamination of food commodities.     

Antimicrobial Efficacy of an Array of Essential Oils Against Lactic Acid Bacteria

The essential oils of clove bud, cinnamon bark and thyme, and their individual compounds including allyl isothiocyanate (AIT), carvacrol, cinnamaldehyde, cinnamic acid, eugenol, and thymol were initially assessed for antimicrobial activity against 9 lactic acid bacteria (LAB) species. Carvacrol and thymol were the most inhibitory with MICs of 0.1% (v/v and w/v, respectively). Cinnamaldehyde, cinnamon bark oil, clove bud oil, eugenol, and thyme oil were moderately inhibitive (MICs = 0.2% v/v), while cinnamic acid required a concentration of 0.5% (w/v). AIT was not effective with MICs in excess of concentrations tested (0.75% v/v). The bactericidal capability of the oil components carvacrol, cinnamaldehyde, eugenol, and thymol were further examined against Pediococcus acidilactici, Lactobacillus buchneri, and Leuconostoc citrovorum. Thymol at 0.1% (w/v) was bactericidal against L. citrovorum (>4‐log reduction), but resulted in a 2‐log CFU/mL reduction against L. buchneri and P. acidilactici. Cinnamaldehyde at 0.2% to 0.25% (v/v) was effective against L. citrovorum, L. buchneri, and P. acidilactici, resulting in a >2‐log reduction. All 3 organisms were susceptible to 0.2% carvacrol with >3‐log reduction observed after exposure for 6 h. Eugenol was the least effective. Concentrations of 0.2% and 0.25% (v/v) were needed to achieve an initial reduction in population, >3‐log CFU/mL after 6 h exposure. However, at 0.2%, P. acidilactici and L. buchneri recovered to initial populations in 48 to 72 h. Results indicate essential oils have the capacity to inactivate LAB that are commonly associated with spoilage of shelf stable low‐acid foods.     

Effects of Essential Oils from Plants on Growth of Food Spoilage Yeasts

Thirty-two essential oils from plants were screened for inhibitory effects on 13 food-spoilage and industrial yeasts. Of these, essential oils of allspice, cinnamon, clove, garlic, onion, oregano, savory, and thyme were most inhibitory. Oils were subsequently tested for their effects on biomass production and pseudomycelium formation of eight genera of yeasts. Garlic oil was a potent inhibitor of yeast growth at concentrations as low as 25 ppm. The oils of onion, oregano and thyme were also strongly inhibitory. Essential oils (100 ppm) had no effect on pseudomycelium production by Candida lipolytica. However, all eight essential oils delayed pseudomycelium formation by Hansenula anomala, whereas six oils stimulated pseudomycelium production by Lodderomyces elongisporus. Cinnamon and clove oils were clearly stimulatory to pseudomycelium production by Saccharomyces cerevisiae.     

The effects of citrus oils and some spices on growth and aflatoxin production by Aspergillus parasiticus NRRL 2999

The inhibitory effect of various concentrations of mint, sage, bay leaves, thyme, aniseed, red pepper and citrus peel oils on the growth of an aflatoxin-producing strain of Aspergillus parasiticus and its production of aflatoxin was studied. Amongst the herbs used, thyme was found a highly effective antifungal agent. Growth and aflatoxin formation was depressed by orange and lemon oils at a concentration of 1.6% through 10 days incubation.     

Determination of Chemical Composition,Total Phenolic,Antimicrobial, and Antioxidant Activities of Echinophora tenuifolia Essential Oil

In this study, the antioxidant and antimicrobial activities, total phenolic content, and essential oil composition of Echinophora tenuifolia L. subsp. sibthorpiana were investigated. The antioxidant activity of investigated essential oil was assessed by ABTS and DPPH assays. DPPH radical scavenging activity expressed by IC₅₀ was 2.84 g/L, whereas the TEAC value determined by ABTS assay was 0.032 g TEAC/kg plant. Total phenol content of essential oil determined by Folin-Ciocalteu method was calculated as 1.32 g GAE/kg plant. The essential oil extracted by hydrodistillation (Clevenger apparatus) was investigated by GC-MS technique and 78 compounds were identified. The main components of essential oils were found to be δ-3-carene (17.93%), p-cymene (8.99%), methyleugenol (16.41%), and α-phellandrene (9.33%). The antimicrobial activity of investigated essential oil was tested using a broth dilution method against 13 bacterial and 2 fungal microorganisms. The lowest minimum inhibitory concentration of essential oil against Bacillus cereus was 62.5 μg/mL while the antifungal activity was greater than 1000 μg/mL for both Candida albicans and Saccharomyces cereviciae. Investigated essential oil has a certain level of antioxidant and antimicrobial effects, which may be attributed to their chemical compounds. The antimicrobial efficiency of essential oil, especially against Bacillus cereus and Staphylocoocus spp., offers its effectiveness to treatment of wound or disease caused by Gram positive bacteria.     

Biocontrol of Gray Mold Disease on Strawberry Fruit by Integration of Lactobacillus plantarum A7 with Ajwain and Cinnamon Essential Oils

This study was conducted to evaluate the efficacy of the Lactobacillus plantarum A7 (L. Plantarum), ajwain and cinnamon essential oils (AO and CO, respectively) in suppressing gray mold rot in strawberry fruit. AO and CO showed over 90% inhibition of radial mycelia growth with lower concentration of the oils per plate for all tested pathogens. Combined application of L. plantarum with AO and CO was tested to assess the possible synergistic effects of these 3 elements on the control of tested plant pathogens. In this case both combinations of L. plantarum + AO and L. plantarum + CO inhibited the mycelia growth of the pathogens completely. Results showed that the combined treatment of strawberry fruits with L. plantarum + AO (50 μL) and L. plantarum + CO (100 μL) resulted in remarkably improved control of Botrytis infections, in comparison with application of L. plantarum or essential oils alone. Quality attributes (that is pH, acidity, vitamin C, and total soluble solid) of the strawberry fruits did not change significantly (P < 0.01) when combination of Lactobacillus and essential oils was used. To the best of our knowledge, this is the first report on the effects of combination of a Lactobacillus as an antagonist bacterium with essential oils to increase the shelf life of strawberry.     

Antifungal activity of essential oils of clove (Syzygium aromaticum) and/or mustard (Brassica nigra) in vapor phase against gray mold (Botrytis cinerea) in strawberries

The search for new alternatives to preserve foods is a growing research field that is of great interest to the food industry. In this study, the inhibitory effect of clove and/or mustard essential oils in vapor phase was evaluated in vitro and in vivo (strawberries) against Botrytis cinerea, a mold highly detrimental to fruits and vegetables. In vitro and in vivo minimum inhibitory concentration (MIC) for clove essential oil was 92.56 μL/L_air, while for mustard, essential oil was 15.42 μL/L_air. As for the combination of tested essential oils the MIC was 46.28 μL/L_air of clove essential oil and 1.93 μL/L_air of mustard essential oil for the in vitro tests, while 11.57 μL/L_air of clove essential oil and 1.93 μL/L_air of mustard essential oil for the in vivo tests. Furthermore, the observed inhibitory effect is due to a synergism of tested essential oils in most of the assessed combinations.Industrial relevanceEffectiveness of vapors of clove and/or mustard essential oils against B. cinerea was demonstrated. The inhibitory activity exhibited by essential oils vapors can be attributed to the major compounds identified in clove and mustard essential oils, inhibiting B. cinerea growth in vitro and in vivo (strawberries). Combinations of clove and mustard essential oils had a synergistic antifungal effect, confirming that the combined use of volatile compounds from essential oils is more effective than its individual application.     

植物复合精油对5种稻谷霉变优势菌株抑菌作用研究

为了研究天然植物精油(百里香、丁香、肉桂)对霉变稻谷的抑菌效果,以5种稻谷霉变优势菌株为受试菌,以霉菌抑菌圈直径大小和最低抑菌浓度（MIC）为指标,通过混料设计方法建立复合精油抑菌模型,结合方差分析得到抑菌效果最佳的植物精油配比。研究结果表明,单一精油抑菌活性对亮白曲霉（A. candidus）,杂色曲霉（A. versicolor）和聚多曲霉（A. sydowii）为肉桂精油>丁香精油>百里香精油;对稻黑孢霉（N. oryzae）为肉桂精油=丁香精油>百里精香油;对布罗克青霉菌（P. brocae）为丁香精油=百里香精油<肉桂精油。当肉桂精油:丁香精油:百里香精油的体积比为55.2%︰26.9%︰17.9%时,3种植物精油对5种菌株抑制效果最佳,复合抑制值大于90.9%。     

Hake proteins edible films incorporated with essential oils: Physical,mechanical, antioxidant and antibacterial properties

Physical, mechanical, antioxidant and antimicrobial properties of hake protein films incorporated with citronella, coriander, tarragon and thyme oils were investigated. Dried hake proteins were solubilized at pH 11.0 (protein concentration in film forming solution ca. 0.9%) and glycerol (59% w/w of protein) and 0.25 ml of each essential oil per gram of protein was added. Films obtained were homogeneous and transparent with a yellowish colour. The addition of the different essential oils reduced the water vapour permeability but increased the solubility of films in water. The amount of protein released from the films depended on the oil added. Among all essential oils incorporated-films the thyme oil films exhibited the lowest mechanical properties (puncture force and elongation at break). Conversely, films with thyme oil added presented the highest inhibition against Shewanella putrefaciens. Antioxidant activity of hake protein films generally increased with the incorporation of essential oils as indicated by DPPH radical scavenging activity and reducing power.