Stéphane Caillet

Mechanism of Action of Spanish Oregano, Chinese Cinnamon, and Savory Essential Oils against Cell Membranes and Walls of Escherichia coli O157:H7 and Listeria monocytogenes

The mechanism of the antimicrobial action of Spanish oregano (Corydothymus capitatus), Chinese cinnamon (Cinnamomum cassia), and savory (Satureja montana) essential oils against cell membranes and walls of bacteria was studied by the measurement of the intracellular pH and ATP concentration, the release of cell constituents, and the electronic microscopy observations of the cells when these essential oils at their MICs were in contact with Escherichia coli O157:H7 and Listeria monocytogenes. E. coli O157:H7 and L. monocytogenes, two pathogenic foodborne bacteria, were used as gram-negative and gram-positive bacterial models, respectively. Treatment with these essential oils at their MICs affected the membrane integrity of bacteria and induced depletion of the intracellular ATP concentration. Spanish oregano and savory essential oils, however, induced more depletion than Chinese cinnamon oil. An increase of the extracellular ATP concentration was observed only when Spanish oregano and savory oils were in contact with E. coli O157:H7 and L. monocytogenes. Also, a significantly higher (P < or = 0.05) cell constituent release was observed in the supernatant when E. coli O157:H7 and L. monocytogenes cells were treated with Chinese cinnamon and Spanish oregano oils. Chinese cinnamon oil was more effective to reduce significantly the intracellular pH of E. coli O157:H7, whereas Chinese cinnamon and Spanish oregano decreased more significantly the intracellular pH of L. monocytogenes. Electronic microscopy observations revealed that the cell membrane of both treated bacteria was significantly damaged. These results suggest that the cytoplasmic membrane is involved in the toxic action of essential oils.

Antimicrobial effects of selected plant essential oils on the growth of a Pseudomonas putida strain isolated from meat

The inhibitory effect of 60 different essential oils was evaluated on a Pseudomonas putida strain of meat origin, associated with meat spoilage. Essential oils were tested at concentrations from 0.003 to 0.8% (wt/vol) to determine minimum inhibitory and maximal tolerated concentrations (MIC and MTC, respectively) using an agar medium culture. Of the 60 samples tested, Corydothymus capitatus essential oil was the most active showing a MIC of 0.025% and a MTC of 0.06%. Seven essential oils (Cinnamomum cassia, Origanum compactum, Origanum heracleoticum, Satureja hortensis, Satureja montana, Thymus vulgaris carvacroliferum, Thymus vulgaris thymoliferum) have shown a strong antimicrobial activity against P. putida with a MIC of 0.05% and a MTC ranging from 0.013% to 0.025%. Ten other oils (Cinnamomum verum (leaf and bark), Eugenia caryophyllus, Cymbopogon martinii var. motia, Cymbopogon nardus, Melaleuca linariifolia, Origanum majorana, Pimenta dioica, Thymus satureoides, Thymus serpyllum) showed a high antimicrobial activity showing a MIC ranging from 0.1% to 0.4%, while the remaining were less active showing a MIC⩾0.8%.

Bioactive Compounds in Cranberries and their Biological Properties

Cranberries are healthy fruit that contribute color, flavor, nutritional value, and functionality. They are one of only three fruits native to America. Over the past decade, public interest for the North American cranberry (Vaccinium macrocarpon) has been rising with reports of their potential health benefits linked to the numerous phytochemicals present in the fruit—the anthocyanins, the flavonols, the flavan-3-ols, the proanthocyanidins, and the phenolic acid derivatives. The presence of these phytochemicals appears to be responsible for the cranberry property of preventing many diseases and infections, including cardiovascular diseases, various cancers, and infections involving the urinary tract, dental health, and Helicobacter pylori-induced stomach ulcers and cancers. Recent years have seen important breakthroughs in our understanding of the mechanisms through which these compounds exert their beneficial biological effects, yet these remain to be scientifically substantiated. In this paper these characteristics, as well as the antioxidant, radical scavenging, antibacterial, antimutagen, and anticarcinogen properties of cranberry major bioactive compounds are explained.

Antimicrobial Effects of Alginate-Based Film Containing Essential Oils for the Preservation of Whole Beef Muscle

Alginate-based edible films containing 1% (wt/vol) essential oils of Spanish oregano, Chinese cinnamon, or savory were immersed in 2% (wt/vol) or 20% (wt/vol) CaCl2 solution and then applied to beef muscle slices to control the growth of Escherichia coli O157:H7 and Salmonella Typhimurium. Whole beef muscle surfaces were inoculated with one of these strains at 10(3) CFU/cm2. During the 5 days of storage, samples of meat were obtained periodically for microbiological analysis. The availability of active compounds from essential oils present in films was evaluated by determination of total phenolic compounds for oregano- and savory-based films and of total aldehydes for cinnamon-based films during storage. After 5 days of storage, films containing oregano or cinnamon essential oils were the most effective against Salmonella Typhimurium regardless of the type of pretreatment used (2 or 20% CaCl2). During the same period, meat inoculated with E. coli O157:H7 and coated with films treated with 2% CaCl2 had significantly fewer bacteria (P < or = 0.05) when oregano-based films were used than when cinnamon- and savory-based films were used. The E. coli O157:H7 concentration was higher at the end of the storage period when films were pretreated with 20% CaCl2. Evaluation of the active compounds in films revealed that availability in oregano- and savory-based films was significantly more important (P < or = 0.05) than that in cinnamon-based films regardless of the type of pretreatment used (2 or 20% CaCl2). At the end of storage, release rates of 40, 60, and 77% were noted in oregano-, savory-, and cinnamon-based films in 2% CaCl2 and rates of 65, 62, and 90% were noted in the same films in 20% CaCl2.

Analyzing cranberry bioactive compounds.

There is a growing public interest for the North American cranberry (Vaccinium macrocarpon) as a functional food because of the potential health benefits linked to phytochemical compounds present in the fruit—the anthocyanin pigments, responsible for its brilliant red color, and other secondary plant metabolites (flavonols, flavan-3-ols, proanthocyanidins, and phenolic acid derivatives). Isolation of these phenolic compounds and flavonoids from a sample matrix is a prerequisite to any comprehensive analysis scheme. By far the most widely employed analytical technique for the characterization of these compounds has been high-performance liquid chromatography(HPLC) coupled with ultraviolet-visible(UV/Vis) and mass spectrometer(MS) detection. This review covers the cranberry major bioactive compounds, the extraction and purification methods, and the analytical conditions for HPLC used to characterize them. Extraction, chromatographic separation and detection strategies, analyte determinations, and applications in HPLC are discussed and the information regarding methods of specific cranberry analyte analyses has been summarized in tabular form to provide a means of rapid access to information pertinent to the reader.

Antimicrobial Effects of Alginate-Based Films Containing Essential Oils on Listeria monocytogenes and Salmonella Typhimurium Present in Bologna and Ham

Bologna and ham slices (300 of each) were inoculated with Salmonella Typhimurium or Listeria monocytogenes at 10(3) CFU/cm(2). Alginate-based edible films that had been immersed in a 2 or 20% (wt/vol) CaC12 solution and contained 1% (wt/ vol) essential oils of Spanish oregano (O; Corydothymus capitatus), Chinese cinnamon (C; Cinnamomum cassia), or winter savory (S; Satureja montana) were then applied to slices to control pathogen growth. On bologna, C-based films pretreated with 20% CaC12 were the most effective against the growth of Salmonella Typhimurium and L. monocytogenes. L. monocytogenes was the more sensitive bacterium to O-, C-, and S-based films. L. monocytogenes concentrations were below the detection level (<10 CFU/ml) after 5 days of storage on bologna coated with O-, C-, or S-based films pretreated with 20% CaCl2. On ham, a 1.85 log CFU/cm2 reduction of Salmonella Typhimurium (P < 0.05) was found after 5 days of storage with C-based films regardless of the type of pretreatment used (2 or 20% CaC12) or when coated with O-based films pretreated with 20% CaCl2. L. monocytogenes was highly resistant in ham, even in the presence of O-, C-, or S-based films. However, C-based films pretreated with 20% CaCl2 were the most effective against the growth of L. monocytogenes. Evaluation of the availability of active compounds in films revealed a significantly higher release of active compounds in C-based films (P < 0.05) regardless of pretreatment or meat tested (bologna or ham). O-based films had the lowest release level of active compounds. The release of active compounds from O- and S-based films pretreated with 20% CaCl2 was faster than that in the same respective films pretreated with 2% CaCl2 regardless of the meat type. C-based film pretreated by immersion in a 20% CaCl2 solution was most efficient against both pathogens, and migration of active compounds was higher in C-based films than in O- and S-based films.

Combined Effects of Antimicrobial Coating, Modified Atmosphere Packaging, and Gamma Irradiation on Listeria innocua Present in Ready-to-Use Carrots (Daucus carota)

The objective of this study was to evaluate the effect of an edible antimicrobial coating combined with modified atmosphere (MA) packaging (60% O2, 30% CO2, and 10% N2) and gamma irradiation on peeled minicarrots inoculated with Listeria innocua. Carrots were inoculated with L. innocua (10(3) CFU/g) and then coated with an antimicrobial coating based on calcium caseinate containing trans-cinnamaldehyde. The same formulation without trans-cinnamaldehyde was used as an inactive coating. Coated and uncoated carrots were packed under the MA or under air, irradiated at 0.25 or 0.5 kGy, and stored at 4 +/- 1 degrees C for 21 days. Samples were evaluated periodically for enumeration of L. innocua. Unirradiated carrots stored under air had the highest concentrations of L. innocua after 21 days of storage: 2.23 CFU/g in the uncoated samples and 2.26 CFU/ g in samples coated with the inactive coating. These results suggest that the inactive coating did not have any antimicrobial effect against L. innocua. However, the addition of the antimicrobial coating resulted in a 1.29-log reduction in the concentration of L. innocua in carrots packed under air after 21 days of storage and a 1.08-log reduction in carrots packed under MA after 7 days of storage. After 7 days of storage, no L. innocua was detected in samples treated at 0.5 kGy under air or in samples treated at 0.25 kGy under MA. A complete inhibition of L. innocua was also observed during all storage periods in uncoated and coated samples treated at 0.5 kGy under MA. These results indicate that the combination of irradiation and MA conditions play an important role in the radiosensitization of L. innocua.

Microbial and sensory quality of marinated and irradiated chicken.

Chicken legs were subjected to two pretreatments (packaged in air or marinated in natural plant extracts and then packaged in air) followed by irradiation (0, 3, or 5 kGy). The control and irradiated chicken legs were stored at 4 degrees C and underwent microbial analysis (mesophilic aerobic plate counts and Salmonella detection) and sensory evaluation at predetermined intervals. Microbial analysis indicated that irradiation had a significant effect (P < or = 0.05) on the mesophilic aerobic plate counts of the poultry. For each treatment, the bacterial growth decreased with an increase of irradiation dose. The marinade had an additive effect with irradiation in reducing bacterial growth and controlling proliferation during storage at 4 +/- 1 degree C. No Salmonella was observed until day 12 in marinated chicken irradiated at 3 kGy and for all experiments with chicken legs stored under air or marinated at 5 kGy. However, Salmonella was found in chicken legs irradiated at 3 kGy in air and in nonirradiated samples. The sensory evaluation indicated a significant (P < or = 0.05) difference in odor and flavor intensities between the irradiated chicken at 5 kGy and the control. No significant difference was found (P > 0.05) between the marinated chicken irradiated at 5 kGy and the control.

Combined effects of coating, modified atmosphere packaging, and gamma irradiation on quality maintenance of ready-to-use carrots (Daucus carota)

The objective of this study was to evaluate the effect of an edible coating combined with modified atmosphere (MA; 60% O2, 30% CO2, and 10% N2) packaging and gamma irradiation on the microbiological stability and physicochemical quality of minimally processed carrots. A coating based on calcium caseinate and whey protein isolates was used. Coated and uncoated peeled minicarrots were packed under the MA or air (78.1% N2, 20.9% O2, and 0.036% CO2), irradiated at 0.5 or 1 kGy, and stored at 4 +/- 1 degrees C for 21 days. Samples were evaluated periodically for aerobic plates counts (APCs) and physicochemical properties (firmness, white discoloration, and whiteness index). Gamma irradiation did not significantly affect the physicochemical properties of the carrots (P > 0.05). Microbiological analysis revealed that for uncoated carrots irradiation at 0.5 and 1 kGy under air and MA reduced the APCs by 3.5 and 4 log CFU/g and by 4 and 4.5 log CFU/g, respectively. For coated carrots, irradiation at 0.5 and 1 kGy under air and MA reduced the APCs by 4 and 4.5 log CFU/g and by 3 and 4.25 log CFU/g, respectively. The coating was able to protect carrots against dehydration during storage under air. Coating and irradiation at 1 kGy were also able to protect carrot firmness during storage under air. MA packaging retarded whitening of uncoated carrots but had a detrimental effect on firmness. The edible coating used in this study did not significantly inhibit (P > 0.05) microbial growth on carrots.

Influence of Antimicrobial Compounds and Modified Atmosphere Packaging on Radiation Sensitivity of Listeria monocytogenes Present in Ready-to-Use Carrots (Daucus carota)

Radiosensitization of Listeria monocytogenes was determined in the presence of trans-cinnamaldehyde, Spanish oregano, winter savory, and Chinese cinnamon on peeled minicarrots packed under air or under a modified atmosphere (60% O2, 30% CO2, and 10% N2). Samples were inoculated with L. monocytogenes HPB 2812 serovar 1/2a (106 CFU/g) and were coated separately with each active compound (0.5%, wt/wt) before being packaged under air or the modified atmosphere and irradiated at doses from 0.07 to 2.4 kGy. Results indicated that the bacterium was more resistant to irradiation under air in the absence of active compound. The dose required to reduce L. monocytogenes population by 1 log CFU (D10) was 0.36 kGy for samples packed under air and 0.17 kGy for those packed under the modified atmosphere. The active compounds evaluated in this study had an effect on the radiation sensitivity of L. monocytogenes on carrots. The most efficient compound was trans-cinnamaldehyde, where a mean 3.8-fold increase in relative radiation sensitivity was observed for both atmospheres compared with the control. The addition of winter savory and Chinese cinnamon produced a similar increase in relative radiation sensitivity but only when samples where packed under modified atmosphere conditions.