Effects of essential oils and oleoresins of plants on ethanol production,respiration and sporulation of yeasts

Ethanol production, respiration, and sporulation of yeasts as effected by essential oils and oleoresins of allspice, cinnamon, clove, garlic, onion, oregano, savory, and thyme were investigated. Essential oils of allspice, cinnamon, and clove had little or no effect on ethanol production by Sacchraromyces cerevisiae. Oils of onion, oregano, savory, and thyme delayed and/or reduced the production of ethanol. Overall, essential oils effectively suppressed ethanol production by Hansenula anomala. At the highest concentrations tested (500 μg/ml), only cinnamon, clove, garlic and thyme oleoresins substantially delayed and/or reduced ethanol production by S. cerevisiae. Most of the essential oils (100 μm/ml) impaired the respiratory activity of S. cerevisiae as evidenced by a reduction in CO₂ production. Thyme oleoresin was the strongest inhibitor. Allspice and garlic oils impaired sporulation by H. anomala. All oils delayed sporulation of Lodderomyces elongisporus).     

Inhibition of Growth and Germination of C. botulinum 33A, 40B, and 1623E by Essential Oil of Spices

Effects of clove, thyme, black pepper, pimenta, origanum, garlic, onion, and cinnamon oils on growth and germination of Clostridium botulinum types 33A, 40B, and 1623E were studied. At 200 ppm, ail oils highly inhibited growth of C. botulinum 33A, 40B, and 1623E. At 10 ppm, inhibitory activity of most oils diminished. By activity on C. botulinum growth, oils could be divided into three categories: (1) very active: cinnamon, origanum, and clove; (2) active: pimenta, and thyme; (3) least active: garlic, onion, and black pepper. Effectiveness on germination was quite different. At 150 and 200 ppm all oils totally prevented germination. At 10 ppm garlic and onion showed higher activity than the others. Spores of 33A were more sensitive than 40B and 1623E.     

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.     

Antibacterial activity of different essential oils obtained from spices widely used in Mediterranean diet

Raw and processed foods are open to contamination during their production, sale and distribution. At present, therefore, a wide variety of chemical preservatives are used throughout the food industry to prevent the growth of food spoiling bacteria. However health and economic considerations have led to a search for alternatives, such as essentials oils that can safely be used as substitutes for fungicides and bactericides to partially or completely inhibit the growth of fungi and bacteria. The aim of this work was to determine the effectiveness of the essentials oils from oregano (Origanum vulgare), thyme (Thymus vulgaris), rosemary (Rosmarinus officinalis), sage (Salvia officinalis), cumin (Cuminum cyminum) and clove (Syzygium aromaticum) on the growth of some bacteria commonly used in the food industry, Lactobacillus curvatus, Lactobacillus sakei, Staphylococcus carnosus and Staphylococcus xylosus or related to food spoilage Enterobacter gergoviae, Enterobacter amnigenus. The agar disc diffusion method was used to determine the antibacterial activities of the oils. All six essential oils analysed had an inhibitory effect on the six tested bacteria. Oregano essential oil showed the highest inhibition effect followed by cumin and clove.     

Inhibitory effect of cinnamon, clove, lemongrass, oregano and palmarose essential oils on growth and fumonisin B1 production by Fusarium proliferatum in maize grain

The effect of cinnamon, clove, oregano, palmarose and lemongrass oils on growth and FB1 production by three different isolates of F. proliferatum in irradiated maize grain at 0.995 and 0.950 aw and at 20 and 30 degrees C was evaluated. The five essential oils inhibited growth of F. proliferatum isolates at 0.995 aw at both temperatures, while at 0.950 aw only cinnamon, clove and oregano oils were effective in inhibiting growth of F. proliferatum at 20 degrees C and none of them at 30 degrees C. Cinnamon, oregano and palmarose oils had significant inhibitory effect on FB1 production by the three strains of F. proliferatum at 0.995 aw and both temperatures, while clove and lemongrass oils had only significant inhibitory effect at 30 degrees C. No differences were found using 500 or 1000 microg essential oil g(-1). At 0.950 aw, none of the essential oils had any significant effect on FB1 production. The results suggest that mainly cinnamon and oregano oils could be effective in controlling growth and FB1 production by F. proliferatum in maize under preharvest conditions.     

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%。     

Effect of essential oils of cinnamon,clove, lemon grass,oregano and palmarosa on growth of and fumonisin B1 production by Fusarium verticillioides in maize

The ability of cinnamon, clove, lemon grass, oregano and palmarosa essential oils to prevent growth of and fumonisin B₁ (FB₁) production by Fusarium verticillioides at different water activity (0.95 and 0.995 a_w) and temperature (20 and 30 °C) levels in irradiated maize grain was evaluated. All the essential oils inhibited growth of F verticillioides isolates under all conditions tested, but FB₁ production was only inhibited at 30 °C and 0.995 a_w. Moreover, stimulation of toxin production was found under certain environmental conditions. None of the essential oils showed a significantly greater ability to inhibit FB₁ production when compared with the others. At 1000 mg essential oil kg^?1 maize the essential oils showed a greater inhibitory effect on growth of F verticillioides than at 500 mg kg^?1, but there was no difference in FB₁ production between the two levels of essential oil. Copyright © 2004 Society of Chemical Industry     

Inhibitory activity of essential oils of garlic and onion against bacteria and yeasts

The essential oils of garlic and onion and their constituent sulfides with three or more sulfur atoms were potent inhibitors of yeast growth. The minimum inhibitory concentrations of garlic oil, onion oil, diallyl trisulfide, diallyl tetrasulfide, and dimethyl trisulfide for all the yeasts tested ranged between 2 and 45 ppm. The oils and their constituent sulfides, however, were only very weakly antibacterial, showing minimum inhibitory concentrations of greater than 300 ppm for most of the bacteria tested. The antiyeast activity of garlic oil and onion oil was storage stable and was not influenced by pH. Film formation on soy sauce by Zygosaccharomyces rouxii SS1 was completely prevented for 30 days by the addition of 30 and 40 ppm of garlic oil and onion oil, respectively.     

鲜切西兰花贮藏期病原菌分离鉴定及植物精油对其抑制效果

以‘冬至绿’西兰花为试材,分离、纯化并鉴定鲜切西兰花贮藏期病原菌,筛选出抑制病原菌效果较好的植物精油。通过分离、纯化及致病性实验得到病原真菌菌株AB01,以病原菌内源转录间隔区序列进行进化树分析,鉴定该病原菌为链格孢菌（Alternaria alternata）。选用肉桂、茴香、牛至、丁香、百里香和香茅6 种精油对AB01进行抑菌性实验,通过体外抑菌和活体抑菌筛选出最优的精油种类及含量。结果发现,肉桂精油对AB01的抑菌效果最好,其最低抑菌浓度和最低杀菌浓度分别为0.05 μL/mL和0.15 μL/mL,其次是茴香和牛至精油。0.15 μL/mL肉桂精油用于鲜切西兰花可有效降低花球黄花率和切面褐变率。     

Two Different Processes to Obtain Antimicrobial Packaging Containing Natural Oils

In this study, antimicrobial packaging materials were developed by incorporating known concentrations (w/w) of essentials oils of oregano (Origanum vulgare) and thyme (Thymus vulgaris) into low-density polyethylene (LDPE), suitable for use as food packaging, via two different methods: ionizing treatment and directly by extrusion. The mechanical, barrier, and antimicrobial properties of the packaging were evaluated against the following foodborne pathogens: Salmonella typhimurium, Listeria monocytogenes, and Escherichia coli O157:H7. The results demonstrate that films developed by extrusion incorporating 4% (w/w) of essential oils had a higher inhibitory effect than those obtained using the ionizing treatment. The packaging developed by extrusion containing 1% (w/w) showed a positive inhibitory effect, while those obtained by the ionizing treatment had no inhibitory effect against any of the test microorganisms. The incorporation of essential oils on the LDPE films generated a plasticizer effect, whereas the ones obtained by means of ionizing treatment did significantly affect the barrier properties of the films. The results of this study showed that plant-derived essential oils could be incorporated in active films for food packaging.     

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.     

Antifungal activity of five different essential oils in vapour phase for the control of Colletotrichum gloeosporioides and Lasiodiplodia theobromae in vitro and on mango

Mango fruit has high commercial value; however, major postharvest losses are encountered throughout the supply chain due to postharvest diseases. These results lead to the search for natural fungicide for postharvest diseases control. The antifungal effects of five essential oils (thyme, clove, cinnamon, anise and vitex) were assessed by disc volatilisation method. Thyme oil vapours at 5 μL per Petriplate, and clove and cinnamon oil at 8 μL per Petriplate showed 100% growth inhibition of mango pathogens in vitro. GC/MS analysis of essential oil showed thymol (23.88), o‐cymol (23.88) and terpinolene (23.88) as the major constituents of thyme oil. Clove and cinnamon oils contain 3‐allyl‐2‐methoxyphenol (37.42%) and benzofuran 3‐methyl (17.97%), respectively. Thyme oil as a fumigant at 66.7 μL L^?1 showed a significant (P < 0.05) inhibition on postharvest pathogens of mango fruits stored at 25 °C for 6 days. Results of our study suggest the possibility of using thyme oil as an alternate natural fungicide to manage postharvest diseases in mango.     

Initial screening for inhibitory activity of essential oils on growth of Fusarium verticillioides, F. proliferatum and F. graminearum on maize-based agar media

An in vitro initial screening of a range of 37 essential oils on inhibition of mycelial growth of Fusarium verticillioides, F. proliferatum and F. graminearum under different temperature (20–30°C) and water activity (a_w) (0.95–0.995) conditions was made. The basic medium was a 3% maize meal extract agar. The maize meal agar was modified with glycerol to a range of water activity conditions and the essential oils were incorporated at different concentrations (0, 500, 1000 μg ml⁻¹). Cinnamon leaf, clove, lemongrass, oregano and palmarosa oils were the products tested suitable for being used as novel preservatives in the control of the three Fusarium species studied. Although water activity was determinant for the growth of the isolates, in general, the preservative effects of the oils were not linked to a_w. However, a trend to a higher inhibition by the oils when a_w was low was observed. Temperature had a minor importance in the inhibitory effect of the preservatives. In vivo studies may be required to confirm the usefulness of the results obtained.     

STABILIZATION OF MEAT LIPIDS WITH GROUND SPICES

The antioxidant efficacy of ground clove, ginger, oregano, rosemary, sage, and thyme was investigated in comminuted pork systems. Spices at 200–2000 ppm levels of addition inhibited the formation of the 2-thiobarbituric acid (TBA) reactive substances (TBARS) by 12–96% over 21-days of storage at 4C. The anti-oxidative effect of spices was concentration-dependent; but close followed by sage and then rosemary was most effective. Ginger and thyme exerted the least effect in prevention of oxidation of meat lipids.     

牛至、香茅、丁香精油化学成分及体外抑菌活性研究

采用滤纸片法、肉汤稀释法和点种法研究3种植物精油及其化学成分对5种病原菌的体外抑菌活性。用气相色谱-质谱联用技术(gas chromatography-mass spectrometer,GC-MS)最佳分析条件对化学活性成分进行鉴定,用峰面积归一法测定各化合物在挥发油中的相对百分含量。结果表明:9.5%的牛至、香茅、丁香精油可将白色葡萄球菌、白色念珠菌全部杀灭;4.75%的牛至、香茅、丁香精油可将金黄色葡萄球菌全部杀灭;4.75%牛至、丁香精油可将大肠埃希菌全部杀灭,但香茅精油杀灭浓度相对较弱;3种精油对铜绿假单胞菌的杀灭浓度均大于9.5%;GC-MS结果显示,牛至精油中主要含有双戊烯(33.93%)、百里香酚(19.12%)、4-萜品醇(14.92%)、α-蒎烯(10.87%);丁香精油主要成分为丁香酚(86.54%),1-石竹烯(10.41%);香茅精油主要包括香茅醛(31.38%)、香叶醇(18.43%);植物精油具有广谱的抑菌活性。     

Antibacterial properties of plant essential oils

Fifty plant essential oils were examined for their antibacterial properties against 25 genera of bacteria. Four concentrations of each oil were tested using an agar diffusion technique. The ten most inhibitory oils were thyme, cinnamon, bay, clove, almond (bitter), lovage, pimento, marjoram, angelica and nutmeg. The most comprehensively inhibitory extracts were angelica (against 25 genera), bay (24), cinnamon (23), clove (23), thyme (23), almond (bitter) (22), marjoram (22), pimento (22), geranium (21) and lovage (20).     

Effect of oregano and thyme essential oils on the microbiological and chemical quality of refrigerated (4 °C) ready‐to‐eat squid rings

The efficacy of oregano and thyme essential oils (OEO and TEO, respectively) in the quality retention of a refrigerated (4 °C) squid (Loligo vulgaris) ring ready‐to‐eat (RTE) product was studied. Essential oils were added at different concentrations to the coating medium during processing. An inhibitory (P < 0.05) effect of OEO on the microbial activity (aerobes, anaerobes, Enterobacteriaceae, psychrotrophs) of the squid rings was observed, with a more pronounced effect as OEO concentration increased. The addition of OEO also led to an inhibitory (P < 0.05) effect on lipid oxidation, as determined by peroxide, thiobarbituric acid‐reactive substance and interaction compound formation; however, no effect (P > 0.05) of the OEO concentrations on lipid oxidation development was detected. The addition of TEO did not lead to an inhibitory effect (P > 0.05) on the microbial activity of the refrigerated RTE squid, although a slight inhibitory (P < 0.05) effect on lipid oxidation was observed in the batches including the higher TEO concentrations.     

Biodegradable gelatin–chitosan films incorporated with essential oils as antimicrobial agents for fish preservation

Essential oils of clove (Syzygium aromaticum L.), fennel (Foeniculum vulgare Miller), cypress (Cupressus sempervirens L.), lavender (Lavandula angustifolia), thyme (Thymus vulgaris L.), herb-of-the-cross (Verbena officinalis L.), pine (Pinus sylvestris) and rosemary (Rosmarinus officinalis) were tested for their antimicrobial activity on 18 genera of bacteria, which included some important food pathogen and spoilage bacteria. Clove essential oil showed the highest inhibitory effect, followed by rosemary and lavender. In an attempt to evaluate the usefulness of these essential oils as food preservatives, they were also tested on an extract made of fish, where clove and thyme essential oils were the most effective. Then, gelatin–chitosan-based edible films incorporated with clove essential oil were elaborated and their antimicrobial activity tested against six selected microorganisms: Pseudomonas fluorescens, Shewanella putrefaciens, Photobacterium phosphoreum, Listeria innocua, Escherichia coli and Lactobacillus acidophilus. The clove-containing films inhibited all these microorganisms irrespectively of the film matrix or type of microorganism. In a further experiment, when the complex gelatin–chitosan film incorporating clove essential oil was applied to fish during chilled storage, the growth of microorganisms was drastically reduced in gram-negative bacteria, especially enterobacteria, while lactic acid bacteria remained practically constant for much of the storage period. The effect on the microorganisms during this period was in accordance with biochemical indexes of quality, indicating the viability of these films for fish preservation.     

Synergistic antifungal effects of thyme essential oil and Lactobacillus plantarum cell‐free supernatant against Penicillium spp. and in situ effects

This study evaluates the synergistic antifungal effects between thyme essential oils and Lactobacillus plantarum cell‐free supernatant (LCFS) against Penicillium spp. and in situ antifungal activity in rice grains. Thyme essential oil and LCFS showed remarkable antifungal activities against Penicillium spp. with the minimum inhibitory concentration (MIC) of 40 and 80 µL/mL, respectively. The analysis of fractional inhibitory concentration (FIC) index showed the antifungal synergism between thyme essential oil and LCFS against Penicillium spp. with FIC index of 0.5. This synergism also resulted in fourfold reduction in their MICs when applied in combination. The antifungal modes of action were characterized by observing the changes in cell membrane permeability and degradation of fungal cell wall. The combination of thyme essential oil and LCFS (2 × MIC of each) showed remarkable in situ antifungal effect and completely inhibit the growth of Penicillium in rice seeds. The results suggested the possible applications of the observed synergism on actual crops.

Practical applications

Essential oils are used as preservative in food industry and high concentration of essential oil is associated with negative organoleptic characteristics. This study presented a novel approach for synergistic antifungal effects by using the combination of thyme essential oil and Lactobacillus plantarum cell‐free supernatant (LCFS) against Penicillium spp. and systematic evaluation of the antifungal effect by using fractional inhibitory concentration (FIC) index method. This approach will be a role model for future research on synergism and overcome the major drawbacks of using live bacteria and the negative effects arising from antimicrobial activities of essential oils.