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.     

Destruction of Listeria monocytogenes in sturgeon (Acipenser transmontanus) caviar by a combination of nisin with chemical antimicrobials or moderate heat

The objective of this study was to investigate the effect of nisin in combination with heat or antimicrobial chemical treatments (such as lactic acid, chlorous acid, and sodium hypochlorite) on the inhibition of Listeria monocytogenes and total mesophiles in sturgeon (Acipenser transmontanus) caviar. The effects of nisin (250, 500, 750, and 1,000 IU/ml), lactic acid (1, 2, and 3%), chlorous acid (134 and 268 ppm), sodium hypochlorite (150 and 300 ppm), and heat at 60 degrees C for 3 min were evaluated for a five-strain mixture of L. monocytogenes and total mesophiles in sturgeon caviar containing 3.5% salt. Selected combinations of these antimicrobial treatments were also tested. Injured and viable L. monocytogenes cells were recovered using an overlay method. Treating caviar with > or =500 IU/ml nisin initially reduced L. monocytogenes by 2 to 2.5 log units. Chlorous acid (268 ppm) reduced L. monocytogenes from 7.7 log units to undetectable (<0.48 log units) after 4 days of storage at 4 degrees C. However, there were no synergistic effects observed for combinations of nisin (500 or 750 IU/ml) plus either lactic acid or chlorous acid. Lactic acid caused a slight reduction (approximately 1 log unit) in the microbial load during a 6-day period at 4 degrees C. Sodium hypochlorite was ineffective at the levels tested. Mild heating (60 degrees C for 3 min) with nisin synergistically reduced viable counts of L. monocytogenes and total mesophiles. No L. monocytogenes cells (<0.48 log units) were recovered from caviar treated with heat and nisin (750 IU/ml) after a storage period of 28 days at 4 degrees C.     

Survival of Listeria monocytogenes Scott A on vacuum-packaged raw beef treated with polylactic acid, lactic acid, and nisin

Low-molecular-weight polylactic acid (LMW-PLA) and lactic acid (LA) were used to inhibit growth of Listeria monocytogenes Scott A on vacuum-packaged beef. Nisin was also used simultaneously as an additional hurdle to the growth of this pathogen. Inoculated beef cubes were immersed in a solution of 2% LMW-PLA, 2% LA, 400 IU/ml of nisin, or combinations of each acid and nisin for 5 min and drip-dried for 15 min. The cubes were then vacuum-packaged and stored at 4 degrees C for up to 42 days. Surface pH values of beef cubes treated with 2% LMW-PLA, the combination of 400 IU/ml of nisin and 2% LMW-PLA (2% NPLA), or 400 IU/ml of nisin alone were significantly reduced from 5.59 to 5.18, 5.01, and 5.19, respectively, whereas those decontaminated with 2% LA or 400 IU/ml of nisin and 2% LA (2% NLA) were significantly decreased from 5.59 to 4.92 and 4.83, respectively, at day 0 (P < or = 0.05). The 2% LMW-PLA, 2% LA, 2% NPLA, 2% NLA, and 400 IU/ml of nisin showed immediate bactericidal effects on L. monocytogenes Scott A (1.22-, 1.56-, 1.57-, 1.94-, and 1.64-log10 reduction, respectively) compared with the initial number of 5.33 log10 CFU/cm2 of the untreated control at day 0 (P < or = 0.05). These treatments, combined with vacuum-packaging and refrigeration temperature, succeeded to inhibit growth of L. monocytogenes during storage up to 42 days. At the end of 42 days, the numbers of L. monocytogenes Scott A remaining viable on these samples were 1.21, 0.36, 2.21, 0.84, and 0.89 log10 CFU/cm2, respectively.     

Evaluation of Antibacterial Activity of Whey Protein Isolate Coating Incorporated with Nisin,Grape Seed Extract,Malic Acid,and EDTA on a Turkey Frankfurter System

ABSTRACT: 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 10⁶ 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 °C for 28 d. The L. monocytogenes population (5.5 log/g) decreased to 2.3 log/g after 28 d at 4 °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 °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.     

Inactivation of barotolerant Listeria monocytogenes in sausage by combination of high-pressure processing and food-grade additives

Food-grade additives were used to enhance the efficacy of high-pressure processing (HPP) against barotolerant Listeria monocytogenes. Three strains of L. monocytogenes (Scott A, OSY-8578, and OSY-328) were compared for their sensitivity to HPP, nisin, tert-butylhydroquinone (TBHQ), and their combination. Inactivation of these strains was evaluated in 0.2 M sodium phosphate buffer (pH 7.0) and commercially sterile sausage. A cell suspension of L. monocytogenes in buffer (10(9) CFU/ml) was treated with TBHQ at 100 ppm, nisin at 100 IU/ml, HPP at 400 MPa for 5 min, and combinations of these treatments. Populations of strains Scott A, OSY-8578, and OSY-328 decreased 3.9, 2.7, and 1.3 log with HPP alone and 6.4, 5.2, and 1.9 log with the HPP-TBHQ combination, respectively. Commercially sterile sausage was inoculated with the three L. monocytogenes strains (10(6) to 10(7) CFU/g) and treated with selected combinations of TBHQ (100 to 300 ppm), nisin (100 and 200 ppm), and HPP (600 MPa, 28 degrees C, 5 min). Samples were enriched to detect the viability of the pathogen after the treatments. Most of the samples treated with nisin, TBHQ, or their combination were positive for L. monocytogenes. HPP alone resulted in a modest decrease in the number of positive samples. L. monocytogenes was not detected in any of the inoculated commercial sausage samples after treatment with HPP-TBHQ or HPP-TBHQ-nisin combinations. These results suggest that addition of TBHQ or TBHQ plus nisin to sausage followed by in-package pressurization is a promising method for producing Listeria-free ready-to-eat products.     

Fate of Listeria monocytogenes inoculated onto the surface of model Turkey frankfurter pieces treated with zein coatings containing nisin, sodium diacetate, and sodium lactate at 4 degrees C

The antimicrobial effects of zein coatings containing nisin, sodium lactate, and sodium diacetate against Listeria monocytogenes on turkey frankfurters at 4 degrees C were determined. Our objectives were to determine whether zein, nisin, lactate, and diacetate alone or in combination could control the growth of L. monocytogenes on full-fat turkey frankfurters at 4 degrees C and to determine whether lactate or diacetate had any synergistic effect on the activity of nisin. Turkey frankfurter pieces surface inoculated with L. monocytogenes strain V7 were treated with zein-ethanol-glycerol (ZEG), zein-propylene-glycol (ZPR), ethanol-glycerol (EG), propylene glycol (PR), nisin (N), sodium lactate (L), or sodium diacetate (D) alone or in combination. Over 28 days, treatment with N or D alone reduced L. monocytogenes counts on frankfurters by 6.6 or 6.3 log CFU/g, respectively. N-D treatment reduced L. monocytogenes by 6 log CFU/g. The zein solvents EG and PR reduced L. monocytogenes by about 5.6 and 5.2 log CFU/g, respectively, similar to the results obtained with ZEG and ZPR, which suggests that zein powder per se had no antimicrobial activity. After 28 days, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, and ZPR-N-D-L yielded no detectable CFU. L alone was ineffective. No synergies were observed. N and D when used singly and the combinations of N-D, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, ZPR-N-D-L, EG, and PR were effective as inhibitors of the growth of recontaminating L. monocytogenes cells on full-fat turkey frankfurters.     

Inhibition of Listeria monocytogenes on the surface of individually packaged hot dogs with a packaging film coating containing nisin

The objective of this study was to determine the effectiveness of packaging films coated with a methylcellulose/hydroxypropyl methylcellulose-based solution containing 10,000, 7,500, 2,500, or 156.3 IU/ml nisin for controlling Listeria monocytogenes on the surfaces of vacuum-packaged hot dogs. Barrier film coated with a methylcellulose/hydroxypropyl methylcellulose-based solution containing nisin or no nisin (control) was heat sealed to form individual pouches. Hot dogs were placed in control and nisin-containing pouches and inoculated with a five-strain L. monocytogenes cocktail (approximately 5 log CFU per package), vacuum sealed, and stored for intervals of 2 h and 7, 15, 21, 28, and 60 d at 4 degrees C. After storage, hot dogs and packages were rinsed with 0.1% peptone water. Diluent was spiral plated on modified oxford agar and tryptic soy agar and incubated to obtain counts (CFU per package). L. monocytogenes counts on hot dogs packaged in films coated with 156.3 IU/ml nisin decreased slightly (approximately 0.5-log reduction) through day 15 of refrigerated storage but was statistically the same (P > 0.05) as hot dogs packaged in films without nisin after 60 d of storage. Packaging films coated with a cellulose-based solution containing 10,000 and 7,500 IU/ml nisin significantly decreased (P < 0.05) L. monocytogenes populations on the surface of hot dogs by greater than 2 log CFU per package throughout the 60-d study. Similar results were observed for hot dogs packaged in films coated with 2,500 IU/ml nisin; however, L. monocytogenes populations were observed to be approximately 4 log CFU per package after 60 d of refrigerated storage from plate counts on tryptic soy and modified oxford agars.     

Nisin treatment to enhance the efficacy of gamma radiation against Listeria monocytogenes on meat

Treatment of meat with gamma radiation for inactivation of foodborne pathogens might cause undesirable quality changes in the product. The objective of the present study was to use nisin for enhancing the lethality of gamma radiation against Listeria monocytogenes, so that moderate doses of radiation can effectively eliminate the pathogen on meat. Cubes of raw meat (10 g each) were inoculated with L. monocytogenes (10(7)CFU/g) and treated with nisin (10(3) IU/g), gamma radiation (0.25 to 1.5 kGy), or combinations of these treatments. Meat was analyzed for L. monocytogenes survivors immediately after treatment and during storage at 4 °C for up to 72 h. Nisin treatment alone inactivated L. monocytogenes by 1.2 log CFU/g. Gamma radiation caused dose-dependent inactivation of the pathogen. Treatment with combinations of nisin and gamma radiation resulted in an additive antimicrobial effect when inoculated meat was tested during the first 24 h and in a synergistic effect when tested after 72 h of storage at 4 °C. When L. monocytogenes was inoculated onto meat at low levels (4×10(3) CFU/g), treated with nisin (10(3) IU/g), and then irradiated (1.5 kGy) and stored at 4 °C for 72 h, the pathogen's most probable number was <0.03/g, indicating that such a combination is potentially effective in eliminating L. monocytogenes in meat.     

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.     

Use of nisin-coated plastic films to control Listeria monocytogenes on vacuum-packaged cold-smoked salmon

Cold-smoked (Salmo salar) salmon samples were surface-inoculated with a cocktail of three nisin-resistant strains of L. monocytogenes (PSU1, PSU2 and PSU21) to a level of approximately 5 x 10(2) or 5 x 10(5) CFU/cm2 of salmon surface. The inoculated smoked salmon samples were vacuum-packaged with control film (no nisin) or nisin-coated plastic films and stored at either 4 or 10 degrees C. When the inoculated smoked salmon samples were packaged with film coated with 2000 IU/cm2 of nisin, a reduction of 3.9 log CFU/cm2 (compared with control) was achieved at either temperature for samples inoculated with 5 x 10(2) CFU/cm(2 of L. monocytogenes after 56 (4 degrees C) and 49 (10 degrees C) days of storage while reductions of 2.4 and 0.7 log CFU/cm2 were achieved for samples inoculated with a high level of L. monocytogenes (5 x 10(5) CFU/cm2) after 58 (4 degrees C) and 43 (10 degrees C) days, respectively. For samples packaged in film coated with 500 IU/cm2 of nisin, reductions of 0.5 and 1.7 log CFU/cm2 were achieved for samples inoculated with a low level of L. monocytogenes (5 x 10(2) CFU/cm2) after 56 (4 degrees C) and 49 (10 degrees C) days of storage while reductions of 1.8 and 0.8 log CFU/cm2 were achieved for samples inoculated with high level of L. monocytogenes after 58(4 degrees C) and 43 (10 degrees C) days, respectively. In addition, nisin inhibited the proliferation of background microbiota on smoked salmon in a concentration-dependent manner at both storage temperatures although the bacteriostatic effect was more pronounced at refrigeration temperature. This work highlights the potential for incorporating nisin into plastic films for enhancing the microbial safety of smoked salmon as well as controlling its microbial spoilage.     

Antimicrobial activity of reuterin in combination with nisin against food-borne pathogens

Antimicrobial activity of reuterin individually or in combination with nisin against different food-borne Gram-positive and Gram-negative pathogens in milk was investigated. Reuterin (8 AU/ml) exhibited bacteriostatic activity against Listeria monocytogenes, whereas its activity was slightly bactericidal against Staphylococcus aureus at 37 degrees C. Higher bactericidal activity was detected against Escherichia coli O157:H7, Salmonella choleraesuis subsp. choleraesuis, Yersinia enterocolitica, Aeromonas hydrophila subsp. hydrophila and Campylobacter jejuni. A significant synergistic effect on L. monocytogenes and a slight additive effect on S. aureus after 24 h at 37 degrees C were observed when reuterin was combined with nisin (100 IU/ml). The combination of reuterin with nisin did not enhance the antimicrobial effect of reuterin against Gram-negative pathogens.     

Effect of inoculum preparation procedure and storage time and temperature on the fate of Listeria monocytogenes on inoculated salami

Although dry/semidry fermented sausages are characterized as being of low-to-moderate risk for human listeriosis on a per-serving and per-annum basis, data are lacking relative to the fate of postprocessing Listeria monocytogenes contamination during storage of such products. This study evaluated the effect of inoculum preparation and storage conditions on the fate of L. monocytogenes on vacuum-packaged salami. Commercially produced salami was sliced and inoculated (4 +/- 1.3 log CFU/ cm2) with one of four types of inocula. All inocula consisted of the same 10-strain L. monocytogenes composite, cultivated as individual strains prior to mixing for inoculation. Active cultures of individual strains were prepared (30 degrees C, 24 h) in either tryptic soy broth (containing 0.25% glucose) plus 0.6% yeast extract (TSBYE), tryptic soy broth without glucose plus 0.6% yeast extract (TSBYE-G), TSBYE-G plus 1% glucose (TSBYE+G), or in TSBYE, and then habituated (7 degrees C, 72 h) in sterile salami homogenate (10% [wt/wt] with distilled water). Inoculated salami slices were vacuum packaged, stored at 4, 12, or 25 degrees C, and analyzed (three samples per treatment in each of two replicates) periodically for surviving bacterial counts. In general, pathogen levels decreased during storage and reached levels below the detection limit (-0.4 log CFU/cm2) between 27 and 90 days of storage, depending on temperature of storage and inoculum type. Death rates (log CFU/cm2/day) were found to increase as storage temperature increased, with the exception of the acid-adapted (TSBYE+G) cells, which decreased more rapidly at 4 degrees C than at 12 or 25 degrees C. The habituated inoculum was inactivated at a faster rate than other inocula at 12 and 25 degrees C, but performed similarly to nonadapted (TSBYE-G) and partially acid-adapted (TSBYE) inocula at 4 degrees C. These data may be used to supplement existing information for use in future risk assessments.     

Control of Listeria monocytogenes on the Surface of Refrigerated, Ready-to-eat Chicken Coated with Edible Zein Film Coatings Containing Nisin and/or Calcium Propionate

ABSTRACT: This study investigated the effect of nisin added to zein film coatings (Z) coated onto ready-to-eat chicken against L. monocytogenes. L. monocytogenes inoculated chicken samples were dipped into Z dissolved in propylene glycol (ZP) or ethanol (ZE), with and without added nisin (N) (1000 IU/g) and/or 1% calcium propionate (CP) then stored at 4 °C or 8 °C for 24 d. After 16 d at 4 °C the growth of L. monocytogenes (6.8 log CFU/g) was suppressed by 4.5 to 5 log CFU/g and at 2.7 log CFU/g counts were maintained at a nondetectable level from day 0 to day 24 with ZEN, ZPNCP, or ZENCP. Zein film coatings with nisin can prevent the growth of L. monocytogenes on ready-to-eat chicken.     

Inhibition of Listeria innocua in hummus by a combination of nisin and citric acid

The effect of nisin or citric acid or combinations of these two inhibitors on the inactivation of a cocktail of three Listeria innocua strains was investigated in a model brain heart infusion (BHI) broth and hummus (chickpea dip). In BHI broth, citric acid had a limited ability to inhibit L. innocua growth. Nisin initially reduced L. innocua concentrations by about 3 log cycles; however, L. innocua reached concentrations similar to those of the control after 5 days at 22 degrees C. In combination, the effects of 500 IU/ml nisin and 0.2% citric acid were synergistic and resulted in complete elimination of L. innocua in the BHI broth. The inhibition of L. innocua by nisin (500 or 1,000 IU/g), citric acid (0.1, 0.2, or 0.3%), or their combinations also was evaluated in hummus. Citric acid alone did not affect L. innocua growth or the aerobic bacterial plate count. A combination of 1,000 IU/g nisin and 0.3% citric acid was somewhat effective (approximately 1.5-log reduction) in controlling the concentration of L. innocua and the aerobic plate count for up to 6 days. This combination also may be useful, in addition to proper hygienic practices, for minimizing the growth of the pathogen Listeria monocytogenes in hummus.     

Heat and acid tolerance of Listeria monocytogenes after exposure to single and multiple sublethal stresses

The majority of published studies on the adaptive heat or acid tolerance response of Listeria monocytogenes have been performed with a single strain exposed to a single adaptation treatment; however, in food ecosystems, microorganisms commonly exist as multi-species communities and encounter multiple stresses, which may result in "stress hardening". Therefore, the present study evaluated the adaptive responses to heat (52, 57 and 63 degrees C) or lactic acid (pH 3.5) of a 10-strain composite of L. monocytogenes meat and human isolates at stationary phase, following exposure to combinations of osmotic (10% NaCl), acidic (pH 5.0 with HCl) and thermal (T; 46 degrees C) stresses, sequentially or simultaneously within 1.5h, in tryptic soy broth with 0.6% yeast extract (TSBYE). All treatments induced adaptive responses on L. monocytogenes at 57 degrees C, while no such cross-protection was observed at 52 and 63 degrees C. Survivor curves at 57 degrees C appeared convex with profound shoulders determined by a Weibull model. The highest thermotolerance was observed after combined exposure to acid and heat shock (pH-T), followed by exposure to osmotic shock, and by the combination of osmotic with heat shock (NaCl-T). Regarding acid tolerance, prior exposure to low pH, pH-T, or a combination of NaCl, pH and T resulted in a marked increase of resistance to pH 3.5, showing concave inactivation curves with tails at higher levels of survivors (log(10)CFU ml(-1)) than the control cultures. The sequence of exposure to sublethal stresses did not affect the thermotolerance of L. monocytogenes, whereas simultaneous exposure to most multiple stresses (e.g., NaCl-pH-T, NaCl-T and NaCl-pH) resulted in higher survivors of L. monocytogenes at pH 3.5 than exposure to the same stresses sequentially. The results indicate that combinations and sequences of sublethal hurdles may affect L. monocytogenes acid and heat tolerance, especially in acidic environments with mild heating or in low moisture environments.     

Inhibition of Listeria monocytogenes in pH-adjusted pasteurized liquid whole egg

Although the transmission of L. monocytogenes to humans via pasteurized egg products has not been documented, L. monocytogenes and other Listeria species have been isolated from commercially broken raw liquid whole egg (LWE) in both the United States and Ireland. Recent Listeria thermal inactivation studies indicate that conventional minimal egg pasteurization processes would effect only a 2.1- to 2.7-order-of-magnitude inactivation of L. monocytogenes in LWE; thus, the margin of safety provided by conventional pasteurization processes is substantially smaller for L. monocytogenes than for Salmonella species (a 9-order-of-magnitude process). The objective of this study was to evaluate the inhibitory effects of nisin on the survival and growth of L. monocytogenes in refrigerated and pH-adjusted (pH 6.6 versus pH 7.5) ultrapasteurized LWE and in a liquid model system. The addition of nisin (1,000 IU/ml) to pH-adjusted ultrapasteurized LWE reduced L. monocytogenes populations by 1.6 to > 3.3 log CFU/ml and delayed (pH 7.5) or prevented (pH 6.6) the growth of the pathogen for 8 to 12 weeks at 4 and 10 degrees C. Bioactive nisin was detected in LWE at both pH values for 12 weeks at 4 degrees C. In subsequent experiments, Listeria reductions of > 3.0 log CFU/ml were achieved within 24 h in both LWE and broth plus nisin (500 IU/ml) at pH 6.6 but not at pH 7.5, and antilisterial activity was enhanced when nisin was added as a solution rather than in dry form.     

Effect of nisin and its combination with sodium chloride on the survival of Listeria monocytogenes added to raw buffalo meat mince

Antilisterial activity of nisin (Nisaplin), alone at concentrations of 400 and 800 IU/g and in combination with 2% sodium chloride was incorporated in raw buffalo meat mince. Samples of the raw meat mince were inoculated with 10(3) colony forming units (cfu)/g of L. monocytogenes and stored at 4°C for 16 days and at 37°C for 36 h. Initial estimates of pH, extract release volume, mesophilic and psychrophilic counts were found to be 5.74, 48 ml, 3.5×10(5) and 1.0×10(5) cfu/g of meat, respectively. The growth of L. monocytogenes in the treated groups was significantly (P<0.05) inhibited compared to the control group. The degree of inhibition increased with increasing concentration of nisin and decreasing storage temperature. Addition of 2% sodium chloride in combination with nisin increased the efficacy of nisin at both storage temperatures. The pH in the treated groups remained significantly lower (P<0.01) than in the control groups at both 4 and 37°C.     

Inhibition of Listeria monocytogenes in cooked ham through active packaging with natural antimicrobials and high-pressure processing

Enterocins A and B and sakacin K at 200 and 2,000 activity units (AU)/cm2, nisin at 200 AU/cm2, 1.8% potassium lactate, and a combination of 200 AU/cm2 of nisin and 1.8% lactate were incorporated into interleavers, and their effectiveness against Listeria monocytogenes spiked in sliced, cooked ham was evaluated. Antimicrobial-packaged cooked ham was then subjected to high-pressure processing (HPP) at 400 MPa. In nonpressurized samples, nisin plus lactate-containing interleavers were the most effective, inhibiting L. monocytogenes growth for 30 days at 6 degrees C, with counts that were 1.9 log CFU/g lower than in the control after 3 months. In the other antimicrobial-containing interleavers, L. monocytogenes did not exhibit a lag phase and progressively grew to levels of about 8 log CFU/g. HPP of actively packaged ham slices reduced Listeria populations about 4 log CFU/g in all batches containing bacteriocins (i.e., nisin, sakacin, and enterocins). At the end of storage, L. monocytogenes levels in the bacteriocin-containing batches were the lowest, with counts below 1.51 log CFU/g. In contrast, HPP moderately reduced L. monocytogenes counts in the control and lactate batches, with populations gradually increasing to about 6.5 log CFU/g at the end of storage.     

Factors affecting the antilisterial effects of nisin in milk

The ability of Listeria monocytogenes to proliferate in milk and the antilisterial activities of nisin are well documented. Although milk fat was reported to reduce the antimicrobial activities of nisin, there is little information on the influence of milk fat on the antilisterial activities of nisin in refrigerated milk, and whether pasteurization and homogenization influence these activities. Fresh, pasteurized, and homogenized milk samples (0.1%, 2.0%, and 3.5% fat) were treated with nisin (0-500 IU/ml) and challenged with 10(4) CFU/ml L. monocytogenes strain Scott A. The organism was most sensitive to nisin in skim milk, showing rapid decline in cell numbers to <10 CFU/ml after 12 days at 5 degrees C following treatment with 250 IU/ml. An initial decline in cell numbers in 2% and whole milk was followed by regrowth of the organism. Loss of the antilisterial effects of nisin was confirmed in homogenized whole milk, whether raw or pasteurized, but not in raw or pasteurized whole milk that was not homogenized. Tween 80, a nonionic emulsifier, partially counteracted the loss of the antilisterial activity of nisin, whereas lecithin, an anionic emulsifier, had no effect. These results demonstrate that the chemical composition and treatment of foods may play an important role in the antilisterial effects of nisin.