Bactericidal activities of plant essential oils and some of their isolated constituents against Campylobacter jejuni,Escherichia coli,Listeria monocytogenes,and Salmonella enterica

An improved method of sample preparation was used in a microplate assay to evaluate the bactericidal activity levels of 96 essential oils and 23 oil compounds against Campylobacter jejuni, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica obtained from food and clinical sources. Bactericidal activity (BA50) was defined as the percentage of the sample in the assay mixture that resulted in a 50% decrease in CFU relative to a buffer control. Twenty-seven oils and 12 compounds were active against all four species of bacteria. The oils that were most active against C. jejuni (with BA50 values ranging from 0.003 to 0.009) were marigold, ginger root, jasmine, patchouli, gardenia, cedarwood, carrot seed, celery seed, mugwort, spikenard, and orange bitter oils; those that were most active against E. coli (with BA50 values ranging from 0.046 to 0.14) were oregano, thyme, cinnamon, palmarosa, bay leaf, clove bud, lemon grass, and allspice oils; those that were most active against L monocytogenes (with BA50 values ranging from 0.057 to 0.092) were gardenia, cedarwood, bay leaf, clove bud, oregano, cinnamon, allspice, thyme, and patchouli oils; and those that were most active against S. enterica (with BA50 values ranging from 0.045 to 0.14) were thyme, oregano, cinnamon, clove bud, allspice, bay leaf, palmarosa, and marjoram oils. The oil compounds that were most active against C. jejuni (with BA50 values ranging from 0.003 to 0.034) were cinnamaldehyde, estragole, carvacrol, benzaldehyde, citral, thymol, eugenol, perillaldehyde, carvone R, and geranyl acetate; those that were most active against E. coli (with BA50 values ranging from 0.057 to 0.28) were carvacrol, cinnamaldehyde, thymol, eugenol, salicylaldehyde, geraniol, isoeugenol, citral, perillaldehyde, and estragole; those that were most active against L monocytogenes (with BA50 values ranging from 0.019 to 0.43) were cinnamaldehyde, eugenol, thymol, carvacrol, citral, geraniol, perillaldehyde, carvone S, estragole, and salicylaldehyde; and those that were most active against S. enterica (with BA50 values ranging from 0.034 to 0.21) were thymol, cinnamaldehyde, carvacrol, eugenol, salicylaldehyde, geraniol, isoeugenol, terpineol, perillaldehyde, and estragole. The possible significance of these results with regard to food microbiology is discussed.     

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.     

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%。     

Antioxidant and Antimicrobial Properties of Selected Spice Extracts

Tinctures of eleven plants used as spices (basil, celery, dill, horsetail, lovage, marjoram, milfoil, oregano, parsley, rosemary, and thyme) were tested for their antiradical properties by means of the DPPH (1,1-diphenyl-2-picrylhydrazyl) assay over a two year period. Seven of these plants (basil, lovage, marjoram, milfoil, oregano, rosemary and thyme) were selected to obtain a mixture, which was tested in situ as antioxidant on vegetable and animal fats by an accelerated oxidation test at 110°C. The herbal extract also showed antimicrobial activity against Staphylococcus aureus (ATCC 25923), Streptococcus pyogenes (ATCC 49399), Escherichia coli (ATCC 25922) and Candida albicans (ATCC 24433).     

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.     

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).     

The analysis of essential oils and extracts (oleoresins) from seasonings--a critical review.

A critical review of the analytical methods employed for the determination of the relevant components of seasonings is presented. Where the available methods were inadequate, new ones have been devised. Particular emphasis has been placed on those methods of analysis that provide a rapid and sufficiently accurate appraisal of seasoning extracts and essential oils from seasonings under routine control laboratory conditions. At the same time, the margin of error of these methods has been determined. The individual seasoning extracts were assessed according to the following criteria: (1) essential oil--cardamom, laurel leaves, cloves, origanum (marjoram), sage, and thyme; (2) essential oil and nonvolatile lipids--dillseed, coriander, caraway, mace, nutmeg, pimento (allspice), and celery seed; (3) essential oil and/or pungent ingredients--capsicum, ginger, and pepper; (4) essential oil and/or coloring matter--turmeric (curcuma) and paprika; (5) essential oil and other components--garlic, onion, and cinnamon.     

Effects of Essential Oil Vapour Treatment on the Postharvest Disease Control and Different Defence Responses in Two Mango (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.) Cultivars

The purpose of this study was to investigate the inhibitory effect of essential oils (thyme, clove and cinnamon) in vapour phase against the major fungal diseases of mango in vitro and in vivo. Thyme oil vapour (5 μL/Petri plate) completely inhibited the mycelial growth of Colletotrichum gloeosporioides and Lasiodiplodia theobromae under in vitro condition. Thyme oil vapour at 66.7 μL L^?1 significantly reduced artificially inoculated C. gloeosporioides and L. theobromae in mangoes for 4 days. GC/MS analysis revealed thymol, eugenol and benzofuran, 3-methyl as the dominant compounds in thyme, clove and cinnamon oils, respectively. The activities of defence and antioxidant enzymes including peroxidase, chitinase, phenylalanine ammonia-lyase, β-1,3-glucanase, catalase and superoxide dismutase were enhanced by thyme oil (66.7 μL L^?1) treatment and also help to maintain the phenolic content. Hence, postharvest thyme oil vapour treatment may prove to be an alternative means of controlling disease in mangoes.     

The analysis of essential oils and extracts (oleoresins) from seasonings ‐ A critical review

A critical review of the analytical methods employed for the determination of the relevant components of seasonings is presented. Where the available methods were inadequate, new ones have been devised. Particular emphasis has been placed on those methods of analysis that provide a rapid and sufficiently accurate appraisal of seasoning extracts and essential oils from seasonings under routine control laboratory conditions. At the same time, the margin of error of these methods has been determined. The individual seasoning extracts were assessed according to the following criteria: (1) essential oil — cardamom, laurel leaves, cloves, origanum (marjoram), sage, and thyme; (2) essential oil and nonvolatile lipids — dillseed, coriander, caraway, mace, nutmeg, pimento (allspice), and celery seed; (3) essential oil and/or pungent ingredients — capsicum, ginger, and pepper; (4) essential oil and/or coloring matter — turmeric (curcuma) and paprika; (5) essential oil and other components — garlic, onion, and cinnamon.     

Efficacy of plant extracts in inhibitingAeromonas hydrophilaandListeria monocytogenesin refrigerated,cooked poultry

Alcohol extracts of angelica root, banana purée, bay, caraway seed, carrot root, clove (eugenol), marjoram, pimento leaf and thyme were applied to cooked chicken to determine their antimicrobial activities against Aeromonas hydrophilaand Listeria monocytogenes.Skinless chicken breast meat was cooked to an internal temperature of 85°C, allowed to cool to c. 5°C, then treated by surface application with plant extracts. Low (10 cfu g⁻¹)or high (10⁵ cfu g⁻¹)populations of A. hydrophilaand L. monocytogeneswere applied and samples were stored at either 5 or 15°C for up to 14 or 7 days, respectively. Eugenol and pimento extracts were most effective in inhibiting growth of both bacteria. A. hydrophilawas the more sensitive to the two treatments, with 4 log₁₀ cfu g⁻¹less growth occurring at 14 days at 5°C on eugenol-treated breast meat than on control samples. These results suggested that plant extracts might be useful as antimicrobials in cooked, ready-to-eat chicken meat.     

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.     

DPPH free-radical scavenging ability, total phenolic content, and chemical composition analysis of forty-five kinds of essential oils

Forty-five kinds of commonly used essential oils were employed to investigate the DPPH (1,1-diphenyl2-picrylhydrazyl) radical scavenging ability and total phenolic content of major chemical compositions. The free-radical scavenging ability and total phenolic content of cinnamon leaf and clove bud essential oils are the best among these essential oils. One-half milliliter of cinnamon leaf and clove bud essential oils (10 mg mL EtOH) are shown to be 96.74% and 96.12% of the DPPH (2.5ml, 1.52 × 10^-4 M) free-radical scavenging ability, respectively. Their EC50 (effective concentrations) are 53 and 36 (μg mL^-1). One milligram per milliliter of cinnamon leaf, clove bud, and thyme red essential oils were shown to be 420, 480, and 270 (mg g^-1 of GAE) of total phenolic content, respectively. Eugenol in cinnamon leaf and clove bud essential oils (82.87% and 82.32%, respectively) were analyzed by GC-MS. It is clear that the amounts of the phenol compounds in essential oils and the DPPH free-radical scavenging ability are in direct proportion.     

Heavy metals in spices commonly consumed in Republic of Korea

Levels of lead (Pb), cadmium (Cd), arsenic (As), and mercury (Hg) were evaluated in 359 samples of commonly consumed spices (cinnamon, parsley, basil, oregano, coriander seed, nutmeg, cumin, bay leaf, fenugreek, rosemary, thyme, fennel, sage, clove, marjoram, tarragon, caraway, dill seed, pepper, and turmeric) from the market in the Republic of Korea. The content of Pb, Cd, and As was assessed by acid wet digestion using a microwave oven and inductively coupled plasma mass spectrometry (ICP-MS). The content of Hg was analysed using a direct mercury analyzer (DMA). Pb, Cd, As, and Hg mean content in spices ranged from 0.039–0.972 mg kg^?1, 0.013–0.315 mg kg^?1, 0.121–0.861 mg kg^?1, and 0.001–0.025 mg kg^?1, respectively.     

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.     

生姜贮藏期病原菌分离鉴定及精油抑菌效果

分离、纯化及鉴定生姜贮藏期病原菌,筛选出对抑制病原菌效果较好的精油并验证该精油抑菌效果。通过分离、纯化及致病性实验得到了病原菌菌株ljr4及ljw2。以病原菌18S rDNA序列进行进化树分析,结果表明该两种病原菌分别属于镰刀菌属及被孢霉属。根据病原菌种属选择了6 种精油进行抑菌性实验,结果发现肉桂精油及百里香精油在用量为2 000 μL/L时对病原菌生长抑制率均为100%。在药敏实验中发现肉桂精油比百里香精油抑菌活性更强。肉桂精油对ljr4、ljw2这2 种病原菌最低抑菌用量分别为64、32 μL/L,最低杀菌用量分别为125、500 μL/L。与未熏蒸生姜相比,在贮藏30 d后肉桂精油熏蒸处理分别使接种ljr4及ljw2的生姜感病率下降了59.33%和47.33%。肉桂精油作为生姜贮藏前的熏蒸剂有较好的应用前景。     

The potential application of plant essential oils as natural food preservatives in soft cheese

Investigations were carried out to assess the efficiency of four plant essential oils; bay, clove, cinnamon and thyme as natural food preservatives. The effect of the plant essential oils at concentrations of 0·1, 0·5 and 1% was studied in low-fat and full-fat soft cheese against Listeria monocytogenes and Salmonella enteritidis at 4° and 10°C respectively, over a 14-day period. The composition of the cheese was shown to be an important factor in determining the effectiveness of the plant essential oils. In the low-fat cheese, all four oils at 1% reduced L. monocytogenes to ≤1·0 log₁₀cfu ml⁻¹. In contrast, in the full-fat cheese, oil of clove was the only oil to achieve this reduction. Oil of thyme proved ineffective against S. enteritidis in the full-fat cheese, yet was equally as effective as the other three oils in the low-fat cheese, reducing S. enteritidis to ≤1·0 log₁₀cfu ml⁻¹from day 4 onwards. It is concluded that selected plant essential oils can act as potent inhibitors of L. monocytogenes and S. enteritidis in a food product.     

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。     

Antimicrobial activity of cinnamon and clove oils under modified atmosphere conditions

Mixtures of cinnamon and clove oils were tested for inhibitory activity against important spoilage microorganism of intermediate moisture foods. Four fungal species (Aspergillus flavus, Penicillium roqueforti, Mucor plumbeus and Eurotium sp.), four yeasts species (Debaryomyces hansenii, Pichia membranaefaciens, Zygosaccharomyces rouxii and Candida lipolytica), and two bacteria species (Staphylococcus aureus and Pediococcus halophilus) inoculated separately on agar plates were sealed in a barrier pouch and exposed to essential oil volatiles under a modified atmosphere of low O2 (<0.05-10%) and high CO2 (20% or 40%), with the balance being N2. A. flavus and Eurotium sp. proved to be the most resistant microorganisms. Cinnamon and clove oils added between 1000 and 4000 microL at a ratio of 1:1 were tested for minimum inhibitory volume (MIV) against molds and yeasts. The gas phase above 1000 microL of the oil mixture inhibited growth of C. lipolytica and P. membranaefaciens; 2000 microL inhibited growth of A. flavus, P. roqueforti, M. plumbeus, Eurotium sp., D. hansenii, and Z. rouxii, while inhibition of A. flavus required the addition of 4000 microL. Higher ratios of cinnamon oil/clove oil were more effective for inhibiting the growth of A. flavus.     

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.     

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.