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.     

Phenolic Profile and Antioxidant Activity of Extracts Prepared from Fermented Heat‐Stabilized Defatted Rice Bran

Heat‐stabilized, defatted rice bran (HDRB) serves as a potential source of phenolic compounds which have numerous purported health benefits. An estimated 70% of phenolics present in rice bran are esterified to the arabinoxylan residues of the cell walls. Release of such compounds could provide a value‐added application for HDRB. The objective of this study was to extract and quantify phenolics from HDRB using fermentation technology. Out of 8 organisms selected for rice bran fermentation, Bacillus subtilis subspecies subtilis had the maximum phenolic release of 26.8 mg ferulic acid equivalents (FAE) per gram HDRB. Response surface methodology was used to further optimize the release of rice bran phenolics. An optimum of 28.6 mg FAE/g rice bran was predicted at 168 h, 0.01% inoculation level, and 100 mg HDRB/mL. Fermentation of HDRB for 96 h with B. subtilis subspecies subtilis resulted in a significant increase in phenolic yield, phenolic concentration, and radical scavenging capacity. Fermented rice bran had 4.86 mg gentistic acid, 1.38 mg caffeic acid, 6.03 mg syringic acid, 19.02 mg (‐)‐epicatechin, 4.08 mg p‐courmaric acid, 4.64 mg ferulic acid, 10.04 mg sinapic acid, and 17.59 mg benzoic acid per 100 g fermented extract compared to 0.65 mg p‐courmaric acid and 0.36 mg ferulic acid per 100 g nonfermented extract. The high phenolic content and antioxidant activity of fermented HDRB extract indicates that rice bran fermentation under optimized condition is a potential means of meeting the demand for an effective and affordable antioxidant.     

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.     

Inhibition of Listeria monocytogenes by Nisin Combined with Grape Seed Extract or Green Tea Extract in Soy Protein Film Coated on Turkey Frankfurters

The objective of this study was to evaluate the inhibitory effect of grape seed extract (GSE), green tea extract (GTE), nisin and their combinations (nisin with either GSE or GTE) against Listeria monocytogenes. The inhibitory effect of these natural compounds was evaluated in phosphate buffer solution (PBS) medium containing approximately 10⁹ colony‐forming units (CFU/mL) of L. monocytogenes. The effectiveness of these compounds in a meat model system was evaluated by surface inoculation (approximately 10⁶ CFU/g) of L. monocytogenes onto turkey frankfurters. The inoculated frankfurters were dipped into soy protein film‐forming solutions with and without the addition of antimicrobial agents (GSE 1% or GTE 1% or nisin 10000 IU or combinations). Samples were stored at either 4 °C or 10 °C. The inhibitory effects of edible coatings were evaluated on a weekly basis for 28 d. The greatest inhibitory effect was observed in the PBS medium containing GSE (1%) and nisin (10000 IU/mL), which caused a 9‐log cycle reduction of L. monocytogenes population after 3 h incubation at 37 °C. In the meat system, the L. monocytogenes population (7.1 CFU/g) was decreased by more than 2 log cycle after 28 d at 4 °C and 10 °C, in the samples containing nisin (10000 IU) combined with either GSE (1%) or GTE (1%). This research has demonstrated that the use of an edible film coating containing both nisin and natural extracts is a promising means of controlling the growth and recontamination of L. monocytogenes on ready‐to‐eat meat products.