In vitro antimicrobial activity of essential oils from aromatic plants against selected foodborne pathogens

The purpose of this study was to examine the effectiveness of selected essential oils for the control of growth and survival of pathogenic microorganisms of significant importance in food hygiene and to determine whether the antimicrobial effect was due to the major compounds of the oils. MIC and MBC were determined by the tube dilution method. Essential oils from Thymus vulgaris from Spain and France, Salvia sclarea, Salvia officinalis, Salvia lavandulifolia, Lavandula latifolia, Lavandula angustifolia, three hybrids of Lavandula latifolia x Lavandula angustifolia (Lavandin 'Super', Lavandin 'Abrialis', and Lavandin 'Grosso'), Rosmarinus officinalis, Hissopus officinalis, and Satureja montana were evaluated. Inhibition ranged from the strong activity of Satureja montana and Thymus vulgaris (France) to no inhibition with Salvia sclarea and Hissopus officinalis for each of the test strains: Salmonella Enteritidis, Salmonella Typhimurium, Escherichia coli O157:H7, Yersinia enterocolitica, Shigella flexneri, Listeria monocytogenes serovar 4b, and Staphylococcus aureus. Because some of the essential oils were highly inhibitory in small quantities to selected pathogenic microorganisms, they may provide alternatives to conventional antimicrobial additives in foods.     

The antimicrobial effect of thyme essential oil, nisin, and their combination against Listeria monocytogenes in minced beef during refrigerated storage

The antimicrobial effect of thyme essential oil (EO) at 0.3%, 0.6%, or 0.9%, nisin at 500 or 1000IU/g, and their combination against Listeria monocytogenes was examined in both tryptic soy broth (TSB) and minced beef meat. Thyme EO at 0.3% possessed a weak antibacterial activity against the pathogen in TSB, whereas at 0.9% showed unacceptable organoleptic properties in minced meat. Thus, only the level of 0.6% of EO was further examined against the pathogen in minced meat. Treatment of minced beef meat with nisin at 500 or 1000IU/g showed antibacterial activity against L. monocytogenes, which was dependent on the concentration level of nisin and the strains used. Treatment of minced beef meat with EO at 0.6% showed stronger inhibitory activity against L. monocytogenes than treatment with nisin at 500 or 1000IU/g. All treatments showed stronger inhibitory activity against the pathogens at 10 degrees C than at 4 degrees C. The combined addition of EO at 0.6% and nisin at 500 or 1000IU/g showed a synergistic activity against the pathogen. Most efficient among treatments was the combination of EO at 0.6% with nisin at 1000IU/g, which decreased the population of L. monocytogenes below the official limit of the European Union recently set at 2logcfu/g, during storage at 4 degrees C.     

The antimicrobial effect of thyme essential oil, nisin and their combination against Escherichia coli O157:H7 in minced beef during refrigerated storage

The antimicrobial effect of thyme essential oil (EO) at supplementation levels of 0.3%, 0.6% or 0.9%, nisin at 500 or 1000 IU/g, and their combination, on Escherichia coli O157:H7 was examined in both tryptic soy broth (TSB) and minced beef meat. EO at 0.3% possessed a weak antibacterial activity against the pathogen in TSB, whereas at 0.9% showed unacceptable organoleptic properties in minced meat. Thus, only the level of 0.6% of EO was further examined against the pathogens in minced meat. Treatment of minced beef meat with EO at 0.6% showed an inhibitory activity against E. coli O157:H7 during storage at 10 °C, but not at 4 °C. Treatment of minced beef meat or TSB with nisin at 500 or 1000 IU/g did not show any antibacterial activity against E. coli O157:H7. The combination of EO at 0.6% and nisin at 500 or 1000 IU/g showed an additive effect against the pathogen, which was higher during storage at 10 °C than at 4 °C.     

肉桂精油抑菌性及其在酱卤牛肉保鲜中的应用

以肉制品中常见的5种食品腐败菌和致病菌为研究对象,重点研究与酱卤肉加工过程相关的不同pH、温度、Na Cl浓度等对肉桂精油抑菌效果的影响,并进行气态防腐试验和酱卤牛肉保鲜实验。结果表明:肉桂精油具有广谱抑菌作用,对受试的5种菌均表现出良好的抑菌作用,其中以枯草芽孢杆菌对肉桂精油最为敏感。肉桂精油与pH存在协同抑菌的作用。在酸性环境中肉桂精油的抑菌性能强烈。肉桂精油与一定浓度范围的Na Cl有协同抑菌效果,且肉桂精油具有良好的热稳定性。气态防腐实验表明:肉桂精油具有良好的挥发性,不与菌体直接接触条件下,在密闭培养皿中对受试菌同样表现出良好的抑菌活性。经肉桂精油浸泡过的酱卤牛肉中微生物的生长受到明显的抑制。     

The effect of dietary oregano essential oil on microbial growth of rabbit carcasses during refrigerated storage

The effect of dietary supplementation with oregano essential oil on microbial growth of rabbit carcasses during refrigerated storage was investigated. A total of 45 weaned rabbits were separated into three equal groups with three subgroups each. One group was given the basal diet and served as control and the other two groups were administered diets supplemented with oregano essential oil at levels of 100 and 200 mg/kg diet, respectively (OR100 and OR200 groups). Total viable counts, Pseudomonas spp., lactic acid bacteria, Brochothrix thermosphacta, Enterobacteriaceae and yeast and mould counts, as well as off-odours and appearance of slime were all assessed on rabbit carcasses stored at 3 ± 1 °C for 12 days. The results showed that performance parameters were not affected (p > 0.05) whereas the dietary supplementation with oregano essential oil resulted in lower (p < 0.05) average microbial counts on the carcasses, compared to controls, throughout storage. Dietary supplementation with oregano essential oil at 200 mg/kg was more effective in inhibiting microbial growth compared with 100 mg/kg. Sensory evaluation scores indicated that the carcasses obtained from OR100 and OR200 groups gave a noticeable putrid odour after days 8 and 10, respectively, whereas the control carcasses developed off-odours after the 6th day of storage. Slime formation in the controls was observed after day 6, while the OR100 and OR200 groups were just beginning to show slime after days 8 and 10, respectively.     

Microbiological and chemical quality of ground beef treated with sodium lactate and sodium chloride during refrigerated storage

The effects of sodium lactate (NaL) and sodium chloride (NaCl), either alone (30 g/kg) or in combination (20+20 g/kg), on the microbiological and chemical quality of raw ground beef during vacuum-packaged storage at 2°C were investigated. The results showed that addition of NaL alone or in combination with NaCl significantly delayed the proliferation of aerobic plate counts, psychrotrophic counts, lactic acid bacteria and Enterobacteriaceae and extended the shelf life of the product up to 15 and 21 days, respectively, versus 8 days only for control. Over the storage time (21 days), NaL maintained the ground beef at almost constant pH, while the pH of control or NaCl-treated samples significantly decreased. Lipid oxidation (TBA value) was not affected by addition of NaL. At storage day 21 however, TBA values of both NaL-treated (0.309) and control (0.318) samples were significantly lower than those of samples treated with NaCl (0.463). The combination of NaCl with NaL significantly reduced the oxidative changes caused by NaCl (0.384 versus 0.463). Therefore, NaL alone or in combination with NaCl could be utilized successfully to reduce the microbial growth, maintain the chemical quality, and extend the shelf life of ground beef during refrigerated storage.     

The effect of dietary oregano essential oil on lipid oxidation in raw and cooked chicken during refrigerated storage

The antioxidative effect of dietary supplementation with oregano essential oil on susceptibility of raw and cooked breast and thigh muscle meat of chickens to lipid oxidation during refrigerated storage for 9 days was investigated. Day-old chickens (n=80) were randomly divided into four groups and fed a basal diet containing 30 mg α-tocopheryl acetate kg(-1) feed as control, or basal diet plus 200 mg α-tocopheryl acetate kg(-1) feed, or basal diet plus 50 or 100 mg oregano essential oil kg(-1) for 38 days prior to slaughter. Lipid oxidation was assessed by monitoring malondialdehyde (MDA) formation in raw and cooked meat during 0, 3, 6 and 9 days of refrigerated storage, using the thiobarbituric acid (TBA) assay and third-order derivative spectrophotometry. Results showed that dietary oregano essential oil supplementation exerted antioxidative effects, the supplementation being most effective in retarding lipid oxidation in stored raw and cooked meat at the 100 mg oregano essential oil kg(-1) feed. However, dietary α-tocopheryl acetate supplementation at 200 mg kg(-1) feed displayed greater antioxidant activity than oregano treatments. Thigh muscle was more susceptible to oxidation compared to breast muscle in all treatments, although the former tissues contained α-tocopherol at markedly higher levels.     

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.     

Composition and antimicrobial properties of Sardinian Juniperus essential oils against foodborne pathogens and spoilage microorganisms

In this work, the chemical compositions and antimicrobial properties of Juniperus essential oils and of their main components were determined. Five berry essential oils obtained from different species of Juniperus growing wild in Sardinia were analyzed. The components of the essential oils were identified by gas chromatography-mass spectrometry (GC-MS) analysis. The antimicrobial activities of the oils and their components against food spoilage and pathogenic microorganisms were determined by a broth microdilution method. The GC-MS analysis showed a certain variability in the concentrations of the main constituents of the oils. Alpha-pinene was largely predominant in the oils of the species J. phoenicea subsp. turbinata and J. oxycedrus. Alpha-pinene and myrcene constituted the bulk (67.56%) of the essential oil of J. communis. Significant quantitative differences were observed for myrcene, delta-3-carene, and D-germacrene. The results of the antimicrobial assay show that the oils of J. communis and J. oxycedrus failed to inhibit any of the microorganisms at the highest concentrations tested (MLC > or = 900 microg/ml), while the oils extracted from J. turbinata specimens were active against fungi, particularly against a strain of Aspergillus flavus (an aflatoxin B1 producer). Of the single compounds tested, delta-3-carene was found to possess the broadest spectrum of activity and appeared to contribute significantly to the antifungal activity observed for J. turbinata oils. This activity may be helpful in the prevention of aflatoxin contamination for many foods.     

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.     

In vitro inhibition of foodborne pathogens by volatile oil and organic extracts of Poncirus trifoliata Rafin. seeds

BACKGROUND: Poncirus trifoliata Rafin. is widely used in oriental medicine because of its beneficiary effects on health. In this study, we examined the chemical compositions of the volatile oil isolated from the seeds of P. trifoliata by hydrodistillation, and tested the efficacy of the oil and various organic extracts as an antibacterial potential against a panel of foodborne pathogens. RESULTS: The hydrodistilled volatile oil was analysed by GC–MS. Thirty‐six compounds representing 90.67% of the total oil were identified, of which veridiflorol, spathulenol, α‐humulene, α‐cadinol, δ‐cadinene, T‐muurolol, hexadecanoic acid, germacrene D, bergaptene and aromadendrene were the major volatile compounds. The oil and the organic extracts of chloroform, ethyl acetate and methanol revealed a remarkable antibacterial effect against the tested pathogens such as Staphylococcus aureus, Bacillus subtilis, Listeria monocytogenes, Pseudomonas aeruginosa, Enterobacter aerogenes, Salmonella typhimurium, S. enteritidis and Escherichia coli. Also, the oil had strong detrimental effect on the viable count of the tested bacteria. CONCLUSION: Our findings demonstrate that the oil and organic extracts derived from P. trifoliata seeds might be a potential source of a preservative for the food or pharmaceutical industries. Copyright © 2009 Society of Chemical Industry     

Evaluation of selected quality parameters to monitor essential oil alteration during storage

Several analytical methods including high-performance liquid chromatography (HPLC) were applied on essential oils to establish a set of suitable quality parameters for monitoring changes during storage. For this purpose, 7 essential oils derived from various plant families were exposed to possible worst-case conditions for up to 3 mo at 38 °C under cool white light in the presence of atmospheric oxygen. After storage and immediate freezing at -80 °C to stop further alterations until analysis, individual physicochemical parameters were assessed and are discussed with regard to the precision of the respective method as well as suitability to monitor aging processes. To determine conductivity and pH values, a work-up of essential oils by water extraction was established. Conductivity, peroxide value as well as pH turned out to be suitable parameters to picture alterations. Additionally, HPLC fingerprints were monitored by diode array detection to track underlying chemical changes in essential oil composition. PRACTICAL APPLICATION: This study evaluates a set of analytical parameters to track changes during essential oil storage. On this basis, quality alterations can be monitored.     

芥末精油对芒果采后病原真菌抑制效果和保鲜效应的研究

以"小腰芒"品种为材料,研究芥末精油熏蒸处理对芒果采后病原菌的抑制效应及对芒果采后保鲜效果。体外实验表明,在室温20～22℃下,芥末熏蒸对炭疽病菌和蒂腐病菌具有较强的抑制活性,0.002mL/L能完全抑制病原菌的生长,且随浓度增大,抑制效应增强。活体实验表明,0.005～0.01mL/L空间熏蒸处理,能明显抑制芒果炭疽病菌和蒂腐病菌在芒果果实上的生长,20℃下6d对照果实整果发病,0.01～0.05mL/L处理的芒果果实没有发病。经芥末精油熏蒸24h,浓度在0.05mL/L以下的芒果常温贮藏12d对其食用品质没有明显的影响。     

Antimicrobial activity of eucalyptus essential oil against Pseudomonas in vitro and potential application in refrigerated storage of pork meat

The objective of this study was to investigate the influence of eucalyptus essential oil (EO) on the Pseudomonas in vitro and application in the pork meat. Eucalyptus oil showed the greatest antimicrobial activity to Pseudomonas among five EOs (tea tree oil, eucalyptus oil, clove oil, mentha oil and garlic oil) in vitro. Eucalyptus oil treatment caused the increasing of membrane permeability and the modifications of ultrastructure and morphology of Pseudomonas spp. and P. aeruginosa. Treatment of refrigerated pork with eucalyptus oil led to a significant decrease P. spp. count, pH, total volatile basic nitrogen and 2‐thiobarbituric acid values, as well as higher sensory scores of custom acceptance than those in control at the end of cold storage. It was conclude that eucalyptus EO is a potential alternative to chemical treatment to control the spoilage and extend the shelf life of refrigerated pork meat.