Bactericidal activities of health-promoting, food-derived powders against the foodborne pathogens Escherichia coli, Listeria monocytogenes, Salmonella enterica, and Staphylococcus aureus.

UNLABELLED: We evaluated the relative bactericidal activities (BA(50) ) of 10 presumed health-promoting food-based powders (nutraceuticals) and, for comparison, selected known components against the following foodborne pathogens: Escherichia coli O157:H7, Salmonella enterica, Listeria monocytogenes, and Staphylococcus aureus. The relative activities were evaluated using quantitative bactericidal activity [(BA(50) value, defined as the percentage of the sample in the assay mixture that resulted in a 50% decrease in colony forming units]. The BA(50) values were determined by fitting the data to a sigmoidal curve by regression analysis using concentration-antimicrobial response data. Antimicrobial activity is indicated by a low BA(50) value; meaning less material is needed to kill 50% of the bacteria. Olive pomace, olive juice powder, and oregano leaves were active against all 4 pathogens, suggesting that they behave as broad-spectrum antimicrobials. All powders exhibited strong antimicrobial activity against S. aureus. The following powders showed exceptionally high activity against S. aureus (as indicated by the low BA(50) values shown in parentheses): apple skin extract (0.002%); olive pomace (0.008%); and grape seed extract (0.016%). Listeria bacteria were also highly susceptible to apple skin extract (0.007%). The most active substances provide candidates for the evaluation of antimicrobial effectiveness in human food and animal feed. PRACTICAL APPLICATION: Plant-derived health-promoting food supplements, high in bioactive compounds, are candidates for use as antimicrobials in food.

The olive compound 4-hydroxytyrosol inactivates Staphylococcus aureus bacteria and Staphylococcal Enterotoxin A (SEA).

The foodborne pathogen Staphylococcus aureus produces the virulent staphylococcal enterotoxin A (SEA), a single chain protein which consists of 233 amino acid residues with a molecular weight of 27078 Da. SEA is a superantigen that is reported to contribute to animal (mastitis) and human (emesis, diarrhea, atopic dermatitis, arthritis, and toxic shock) syndromes. Changes in the native structural integrity may inactivate the toxin by preventing molecular interaction with cell membrane receptor sites of their host cells. In the present study, we evaluated the ability of the pure olive compound 4-hydroxytyrosol and a commercial olive powder called Hidrox-12, prepared by freeze-drying olive juice, to inhibit S. aureus bacteria and SEA's biological activity. Dilutions of both test substances inactivated the pathogens. Two independent cell assays (BrdU incorporation into newly synthesized DNA and glycyl-phenylalanyl-aminofluorocoumarin proteolysis) demonstrated that the olive compound 4-hydroxytyrosol also inactivated the biological activity of SEA at concentrations that were not toxic to the spleen cells. However, efforts to determine inhibition of the toxin by Hidrox-12 were not successful because the olive powder was cytotoxic to the spleen cells at concentrations found to be effective against the bacteria. The results suggest that food-compatible and safe antitoxin olive compounds can be used to inactivate both pathogens and toxins produced by the pathogens. Practical Application: The results of this study suggest that food-compatible and safe antitoxin olive compounds can be used to reduce both pathogens and toxins produced by the pathogens in foods.

Mechanical, barrier, and antimicrobial properties of apple puree edible films containing plant essential oils.

Edible films, as carriers of antimicrobial compounds, constitute an approach for incorporating plant essential oils (EOs) onto fresh-cut fruit surfaces. The effect against Escherichia coli O157:H7 of oregano, cinnamon, and lemongrass oils in apple puree film-forming solution (APFFS) and in an edible film made from the apple puree solution (APEF) was investigated along with the mechanical and physical properties of the films. Bactericidal activities of APFFS, expressed as BA50 values (BA50 values are defined as the percentage of antimicrobial that killed 50% of the bacteria under the test conditions) ranged from 0.019% for oregano oil to 0.094% for cinnamon oil. Oregano oil in the apple puree and in the film was highly effective against E. coli O157:H7. The data show that (a) the order of antimicrobial activities was oregano oil > lemongrass oil > cinnamon oil and (b) addition of the essential oils into film-forming solution decreased water vapor permeability and increased oxygen permeability, but did not significantly alter the tensile properties of the films. These results show that plant-derived essential oils can be used to prepare apple-based antimicrobial edible films for various food applications.

Antimicrobial activities of tea catechins and theaflavins and tea extracts against Bacillus cereus.

We evaluated the antimicrobial activities of seven green tea catechins and four black tea theaflavins, generally referred to as flavonoids, as well as the aqueous extracts (infusions) of 36 commercial black, green, oolong, white, and herbal teas against Bacillus cereus (strain RM3190) incubated at 21 degrees C for 3, 15, 30, and 60 min. The results obtained demonstrate that (i) (-)-gallocatechin-3-gallate, (-)-epigallocatechin-3-gallate, (-)-catechin-3-gallate, (-)-epicatechin-3-gallate, theaflavin-3, 3'-digallate, theaflavin-3'-gallate, and theaflavin-3-gallate showed antimicrobial activities at nanomolar levels; (ii) most compounds were more active than were medicinal antibiotics, such as tetracycline or vancomycin, at comparable concentrations; (iii) the bactericidal activities of the teas could be accounted for by the levels of catechins and theaflavins as determined by high-pressure liquid chromatography; (iv) freshly prepared tea infusions were more active than day-old teas; and (v) tea catechins without gallate side chains, gallic acid and the alkaloids caffeine and theobromine also present in teas, and herbal (chamomile and peppermint) teas that contain no flavonoids are all inactive. These studies extend our knowledge about the antimicrobial effects of food ingredients.

Antibacterial activities of plant essential oils and their components against Escherichia coli O157:H7 and Salmonella enterica in apple juice.

We evaluated 17 plant essential oils and nine oil compounds for antibacterial activity against the foodborne pathogens Escherichia coli O157:H7 and Salmonella enterica in apple juices in a bactericidal assay in terms of % of the sample that resulted in a 50% decrease in the number of bacteria (BA(50)). The 10 compounds most active against E. coli (60 min BA(50) range in clear juice, 0.018-0.093%) were carvacrol, oregano oil, geraniol, eugenol, cinnamon leaf oil, citral, clove bud oil, lemongrass oil, cinnamon bark oil, and lemon oil. The corresponding compounds against S. enterica (BA(50) range, 0.0044-0.011%) were Melissa oil, carvacrol, oregano oil, terpeineol, geraniol, lemon oil, citral, lemongrass oil, cinnamon leaf oil, and linalool. The activity (i) was greater for S. enterica than for E. coli, (ii) increased with incubation temperature and storage time, and (iii) was not affected by the acidity of the juices. The antibacterial agents could be divided into two classes: fast-acting and slow-acting. High-performance liquid chromatography analysis showed that the bactericidal results are related to the composition of the oils. These studies provide information about new ways to protect apple juice and other foods against human pathogens.

Antibacterial activities of phenolic benzaldehydes and benzoic acids against Campylobacter jejuni, Escherichia coli, Listeria monocytogenes, and Salmonella enterica.

We evaluated the bactericidal activities of 35 benzaldehydes, 34 benzoic acids, and 1 benzoic acid methyl ester against Campylobacter jejuni, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica when these compounds were substituted on the benzene ring with 0, 1, 2, or 3 hydroxy (OH) and/or methoxy (OCH3) groups in a pH 7.0 buffer. Dose-response plots were used to determine the percentage of the sample that induced a 50% decrease in CFU after 60 min (BA50). Of the 70 compounds tested, 24 were found to be active against all four pathogens, and additional 4, 10, and 12 were found to be active against three, two, and one of the pathogens, respectively. C. jejuni was approximately 100 times as sensitive as the other three pathogens. The 10 compounds that were most active against the four pathogens (with average BA50 values ranging from 0.026 to 0.166) and are candidates for studies of activity in foods or for disinfections were 2,4,6-trihydroxybenzaldehyde, 2,5-dihydroxybenzaldehyde, 2,3,4-trihydroxybenzaldehyde, 2-hydroxy-5-methoxybenzaldehyde, 2,3-dihydroxybenzaldehyde, 2-hydroxy-3-methoxybenzaldehyde, 4-hydroxy-2,6-dimethoxybenzaldehyde, 2,5-dihydroxybenzaldehyde, 2,4-dihydroxybenzaldehyde. and 2-hydroxybenzaldehyde. Comparison of the chemical structures of the test compounds and their activities revealed that (i) the aldehyde (CHO) group was more active than the carboxyl (COOH) group whether or not OH groups were present; (ii) compounds were most active with trisubstituted OH > disubstituted OH > monosubstituted OH; (iii) for disubstituted derivatives, 2-OH enhanced activities were exhibited by benzaldehyde but not by benzoic acid; (iv) compounds were more active with OH than with OCH3, irrespective of the position of substitution on the benzene ring; (v) compounds with mixed OH and OCH3 groups exhibited variable results, i.e., in some cases OCH3 groups enhanced activity and in other cases they did not; (vi) methoxybenzoic acids were largely inactive; and (vii) gallic acid was 20 times as active against S. enterica at pH 7.0 as it was at pH 3.7, suggesting that the ionization of its OH groups may enhance bactericidal activity.

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.