Antilisterial activity and consumer acceptance of irradiated chicken breast meat vacuum-infused with grape seed and green tea extracts and tartaric acid.

Contamination of poultry with pathogenic bacteria contributes to human foodborne disease, causes damage to industry brand names, and has a significant economic impact on the food industry in the form of both damage to industry brand names and losses associated with recalls. Irradiation is a safe and effective means of decontaminating poultry products, but the maximum dose strengths allowed negatively impact poultry sensory quality characteristics. The 1st objective of this study was to investigate the potential interactive inhibitory effects of natural antimicrobials as components of a vacuum-marination in addition to various dose levels of irradiation. Tartaric acid (TA) at 2 levels and grape seed (GS) and green tea (GT) extracts were combined, vacuum-infused into chicken breast fillets, and irradiated at 1, 2, and 3 kGy by electron beam irradiation. The 2nd objective was to use a consumer test group to evaluate TA and plant extract infusion into chicken breast fillets with and without irradiation at 2 kGy on overall impression, flavor, texture, appearance, and tenderness. The results showed that samples vacuum-infused with TA at 37.5 and 75.0 mM and irradiated at 1 kGy significantly reduced Listeria monocytogenes (L.m.) levels by 2 and 3 log CFU/g compared to the control after 12 d of refrigerated storage. Vacuum-infusion of TA at 37.5 and 75.0 mM at 2 and 3 kGy irradiation, reduced L.m. to near nondetectable levels. The addition of TA and GS and GT to chicken breast fillets with and without irradiation did not significantly impact consumer preference, tenderness, appearance, or flavor. The addition of tartaric acid and natural plant extracts to chicken marinades could contribute to the prevention of L.m. contamination.

Ciprofloxacin-sensitive and ciprofloxacin-resistant Campylobacter jejuni are equally susceptible to natural orange oil-based antimicrobials.

A total of 10 ciprofloxacin-sensitive (ciprofloxacin minimum inhibitory concentration, MIC < 0.5 micro g/ml) and 10 ciprofloxacin-resistant (MIC 16 to 32 micro g/ml) presumptive C. jejuni were further characterized and evaluated for their inhibition by natural orange oil fractions. Partial species identification was performed by using a hippuricase gene-based polymerase chain reaction (PCR) assay. One of the isolates appeared to be atypical and failed to hydrolyze hippurate. Of the ciprofloxacin-resistant C. jejuni isolates tested, six were found to have their quinolone resistance determined by a C --> T mutation in codon 86 of gyrA. Both groups of ciprofloxacin-sensitive and -resistant C. jejuni isolates were most susceptible to cold-pressed terpeneless Valencia orange oil (C4) which yielded inhibition zones from 44.0 +/- 1.4 to 80 +/- 0.0 mm. Less inhibitory responses were recorded for 5-fold concentrated Valencia orange oil (C3) and distilled d-limonene (C7) which exerted similar effects on both ciprofloxacin-sensitive and -resistant C. jejuni isolates. In general, ciprofloxacin-resistant and -sensitive C. jejuni isolates were equally susceptible to the respective orange oil fractions.

Effect of organic acids and plant extracts on Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium in broth culture model and chicken meat systems.

Foodborne illness due to consumption of products contaminated with Salmonella Typhimurium (S.T.), Listeria monocytogenes (L.m.), and Escherichia coli O157:H7 (E.c.) results in many deaths and significant economic losses each year. In this study, acetic (AA), citric acid (CA), lactic acid (LA), malic acid (MA), and tartaric acid (TA) and grape seed (GS), green tea (GT), bitter melon seed (BMS), rasum, and fenugreek (FG) extracts were investigated as inhibitors against S.T., L.m., and E.c. in both broth-culture and meat systems. Brain Heart Infusion solutions containing 18.7, 37.5, and 75.0 mM organic acids and 5, 10, 20, and 40 mg/mL extracts were challenged with approximately log 6 CFU/mL S.T., L.m., and E.c. A pH-adjusted control was included to determine pH effect on exhibited antibacterial activity. For the meat system, 1 to 2 g chicken breast pieces were vacuum-infused with CA/MA/TA acid at 75 and 150 mM and GS and GT at 3000, 6000, and 9000 ppm in a partial factorial arrangement. GT and GS showed considerable activity in broth-culture. All organic acids were effective in broth-culture at 75 mM after 24 h (P < 0.05). CA and TA were effective at 37.5 mM. CA/MA/TA at 150.0 mM were the most effective in the meat system, reducing E.c., L.m. and S.T. by >5, >2, and 4-6 log CFU/g, respectively, although all organic acids showed some antibacterial activity at 75.0 and 150.0 mM. These findings demonstrate the effectiveness of organic acids and plant extracts in the control of S.T., L.m., and E.c. O157:H7.

Evaluation of antibacterial activity of whey protein isolate coating incorporated with nisin, grape seed extract, malic acid, and EDTA on a Turkey frankfurter system.

The effectiveness of whey protein isolate (WPI) coatings incorporated with grape seed extract (GSE), nisin (N), malic acid (MA), and ethylenediamine tetraacetic acid (EDTA) and their combinations to inhibit the growth of Listeria monocytogenes, E. coli O157:H7, and Salmonella typhimurium were evaluated in a turkey frankfurter system through surface inoculation (approximately 10(6) CFU/g) of pathogens. The inoculated frankfurters were dipped into WPI film forming solutions both with and without the addition of antimicrobial agents (GSE, MA, or N and EDTA, or combinations). Samples were stored at 4 degrees C for 28 d. The L. monocytogenes population (5.5 log/g) decreased to 2.3 log/g after 28 d at 4 degrees C in the samples containing nisin (6000 IU/g) combined with GSE (0.5%) and MA (1.0%). The S. typhimurium population (6.0 log/g) was decreased to approximately 1 log cycles after 28 d at 4 degrees C in the samples coated with WPI containing a combination of N, MA, GSE, and EDTA. The E. coli O157:H7 population (6.15 log/g) was decreased by 4.6 log cycles after 28 d in samples containing WPI coating incorporated with N, MA, and EDTA. These findings demonstrated that the use of an edible film coating containing nisin, organic acids, and natural extracts is a promising means of controlling the growth and recontamination of L. monocytogenes, S. typhimurium, and E. coli O157:H7 in ready-to-eat poultry products.

Transmission electron microscopy study of Listeria monocytogenes treated with nisin in combination with either grape seed or green tea extract.

The objective of this study was to use transmission electron microscopy to investigate the morphological changes that occurred in Listeria monocytogenes cells treated with grape seed extract (GSE), green tea extract (GTE), nisin, and combinations of nisin with either GSE or GTE. The test solutions were prepared with (i) 1% GSE, 1% GTE, 6,400 IU of nisin, and the combination of these dilutions with nisin or with (ii) the pure major phenolic constituents of GSE (0.02% epicatechin plus 0.02% catechin) or GTE (0.02% epicatechin plus 0.02% caffeic acid) and their combinations with 6,400 IU of nisin in tryptic soy broth with 0.6% yeast extract (TSBYE). Test solutions were inoculated with L. monocytogenes at approximately 10(6) CFU/ml and incubated for 3 or 24 h at 37 degrees C. After 3 h of incubation, cells were harvested and evaluated under a transmission electron microscope (JEOL-100 CX) operating at 80 kV (50,000X). Microscopic examination revealed an altered cell membrane and condensed cytoplasm when L. monocytogenes cells were exposed to a combination of nisin with either GSE or GTE or to pure compounds of the major phenolic constituents in combination. After 24 h of incubation at 37 degrees C, the combinations of nisin with GSE and nisin with GTE reduced the L. monocytogenes population to undetectable levels and 3.7 log CFU/ml, respectively. These observations indicate that the combination of nisin with either GSE or GTE had a synergistic effect, and the combinations of nisin with the major phenolic constituents were most likely associated with the L. monocytogenes cell damage during inactivation in TSBYE at 37 degrees C.

Inhibition of Listeria monocytogenes using nisin with grape seed extract on turkey frankfurters stored at 4 and 10 degrees C.

Recontamination of cooked ready-to-eat (RTE) chicken and beef products with Listeria monocytogenes has been a major safety concern. Natural antimicrobials in combinations can be an alternative approach for controlling L. monocytogenes. Therefore, the objectives of this study were to evaluate the inhibitory activities against L. monocytogenes of nisin (6,400 IU/ ml), grape seed extract (GSE; 1%), and the combination of nisin and GSE both in tryptic soy broth with 0.6% yeast extract (TSBYE) and on the surface of full-fat turkey frankfurters. TSBYE was incubated at 37 degrees C for 72 h and turkey frankfurters at 4 or 10'C for 28 days. Inocula were 6.7 or 5 log CFU per ml or g for TSBYE or frankfurters, respectively. After 72 h in TSBYE, nisin alone did not show any inhibitory activity against L. monocytogenes. The combination of nisin and GSE gave the greatest inhibitory activity in both TSBYE and on turkey frankfurters with reductions of L. monocytogenes populations to undetectable levels after 15 h and 21 days, respectively. This combination of two natural antimicrobials has the potential to control the growth and recontamination of L. monocytogenes on RTE meat products.

Does soil CO2 efflux acclimatize to elevated temperature and CO2 during long-term treatment of Douglas-fir seedlings?

We investigated the effects of elevated soil temperature and atmospheric CO2 on soil CO2 efflux (SCE) during the third and fourth years of study. We hypothesized that elevated temperature would stimulate SCE, and elevated CO2 would also stimulate SCE with the stimulation being greater at higher temperatures. The study was conducted in sun-lit controlled-environment chambers using Douglas-fir (Pseudotsuga menziesii) seedlings grown in reconstructed litter-soil systems. We used a randomized design with two soil temperature and two atmospheric CO2 treatments. The SCE was measured every 4 wk for 18 months. Neither elevated temperature nor CO2 stimulated SCE. Elevated CO2 increased the temperature sensitivity of SCE. During the winter, the relationship between SCE and soil moisture was negative but it was positive during the summer. The seasonal patterns in SCE were associated with seasonal changes in photosynthesis and above-ground plant growth. SCE acclimatized in the high-temperature treatment, probably because of a loss of labile soil carbon. Elevated CO2 treatment increased the temperature sensitivity of SCE, probably through an increase in substrate availability.

Identification and characterization of two bacteriocin-producing bacteria isolated from garlic and ginger root.

Two bacteriocin-producing bacterial strains were isolated from garlic and ginger root by the agar overlay method. The bacteria were identified by 16S rRNA sequence analyses and fermentation patterns as Leuconostoc mesenteroides (garlic isolate) and Lactococcus lactis (ginger isolate). The bacteriocins were assigned the names leucocin BC2 and lactocin GI3, respectively. Physiochemical properties and antimicrobial spectra of the bacteriocins were determined by the spot-on-lawn method. Both bacteriocins were inhibited by proteolytic enzymes. Leucocin BC2 exhibited a narrow antimicrobial spectrum, inhibiting only Bacillus, Enterococcus, and Listeria species. Lactocin GI3 had a broader spectrum, inhibiting Bacillus, Clostridium, Listeria, Enterococcus, Leuconostoc, Pediococcus, and Staphylococcus species. Both bacteriocins remained active when heated at 90 degrees C for 15 min or 120 degrees C for 20 min. Leucocin BC2 assayed at 37 degrees C showed an inhibitory activity of 1,600 AU/ml, whereas at 8 degrees C the activity was 12,800 AU/ml. Conversely, lactocin GI3 activity was the same at both assay temperatures. Both bacteriocins remained active over a pH range of 2.0 to 9.0 and in various organic solvents. The activity of leucocin BC2 was increased when treated with 0.5% acetic acid and 0.5% lactic acid, whereas lactocin GI3 activity was decreased with either acid. The molecular mass values were 3.7 kDa for leucocin BC2 and 3.9 kDa for lactocin GI3. These results show that the inhibitory substances produced by the bacteria isolated from garlic and ginger are bacteriocins that appear to be different in some characteristics from previously reported bacteriocins.

Detection and characterization of a bacteriocin produced by Lactococcus lactis subsp. cremoris R isolated from radish.

Bacteria isolated from radish were identified as Lactococcus lactis subsp. cremoris R and their bacteriocin was designated lactococcin R. Lactococcin R was sensitive to some proteolytic enzymes (proteinase-K, pronase-E, proteases, pepsin, alpha-chymotrypsin) but was resistant to trypsin, papain, catalase, lysozyme and lipase, organic solvents, or heating at 90 degrees C for 15, 30 and 60 min, or 121 degrees C for 15 min. Lactococcin R remained active after storage at -20 and -70 degrees C for 3 months and after exposure to a pH of 2-9. The molecular weight of lactococcin R was about 2.5 kDa. Lactococcin R was active against many food-borne pathogenic and food spoilage bacteria such as Clostridium, Staphylococcus, Listeria, Bacillus, Micrococcus, Enterococcus, Lactobacillus, Leuconostoc, Streptococcus and Pediococcus spp., but was not active against any Gram-negative bacteria. Lactococcin R was produced during log phase and reached a maximum activity (1600 AU ml-1) at early stationary phase. The highest lactococcin R production was obtained in MRS broth with 0.5% glucose, at 6.5-7.0 initial pH values, 30 degrees C temperature and 18-24-h incubation times. Lactococcin R adsorbed maximally to its heat-killed producing cells at pH 6-7 (95%). Crude lactococcin R at 1280 AU ml-1 was bactericidal, reducing colony counts of Listeria monocytogenes by 99.98% in 3 h. Lactococcin R should be useful as a biopreservative to prevent growth of food-borne pathogenic and food spoilage bacteria in ready-to-eat, dairy, meat, poultry and other food products. Lactococcin R differs from nisin in having a lower molecular weight, 2.5 kDa vs 3.4 kDa, and in being sensitive to pepsin and alpha-chymotrypsin to which nisin is resistant.

Death of Salmonella typhimurium and Escherichia coli in the presence of freshly reconstituted dehydrated garlic and onion.

The kinetics of the decline of populations of Salmonella typhimurium inoculated into freshly reconstituted dehydrated onion and garlic powders was studied. Measurable bactericidal activity was observed for onion and garlic concentrations of 1 and 5% (w/v), respectively, with maximal death rates occurring for concentrations of 5 and 10%. At these concentrations, the decimal reduction times were 1.1 and 1.2 hr, respectively, for resting cell cultures and 1.8 and 2.1 hr, respectively, for growing cultures. Of the major volatile aliphatic disulfide compounds of onions, n-propyl allyl and di-n-propyl, at concentrations of 0.1%, showed a comparable activity against resting cells but only a bacteriostatic effect toward actively growing cultures, which overcame this effect in 2 to 6 hr. At comparable concentrations, growing cultures of Escherichia coli were as susceptible to garlic, but apparently more resistant to onion, than were those of S. typhimurium.