Nisin in milk sensitizes Bacillus spores to heat and prevents recovery of survivors.

Decimal reduction times (D values) were determined for Bacillus cereus T spores and B. stearothermophilus ATCC 12980 spores in skim milk supplemented with various concentrations (0, 2,000, and 4,000 IU/ml) of the bacteriocin nisin by using an immersed, sealed capillary tube procedure. For both organisms, the addition of nisin lowered the apparent D values. For B. cereus, the addition of 2,000 IU of nisin per ml to skim milk before heating significantly (P< or =0.05) lowered the apparent D value compared to the control treatment. The D values at 97 degrees C were 7.0, 4.8, and 4.7 min for the control and 2,000- and 4,000-IU/ml nisin treatments, respectively. At 103 degrees C, the D values were 1.5, 0.85, and 0.88 min for the control and 2,000-and 4,000-IU/ml nisin treatments. When calculated across both nisin treatments, the mean reductions in apparent D values at 97, 100, and 103 degrees C due to addition of nisin in comparison to the controls were 32, 20, and 42%, respectively. The zD values for B. cereus ranged from 8.0 to 8.9 degrees C. With B. stearothermophilus, the apparent D values at 130 degrees C were reduced by 13 and 21% respectively, because of the presence of 2,000 or 4,000 IU of nisin per ml. The D values were 16.0, 13.8, and 12.5 s for the control and 2,000- and 4,000-IU/ml nisin treatments, respectively. There was a significant (P< or =0.05) decrease in the apparent D value between the control and 4,000-IU/ml treatment. Overall, log populations of survivors for B. stearothermophilus compared to the control were lower at any given sampling time due to the presence of nisin. The results of these studies suggest that spore control is likely due to enhanced sensitivity of spores to heat and the presence of residual nisin in the recovery medium that could prevent outgrowth of survivors.     

Postprocessing antimicrobial treatments to control Listeria monocytogenes in commercial vacuum-packaged bologna and ham stored at 10 degrees C

The antilisterial effect of chemical dipping solutions on commercial bologna and ham slices, inoculated (3 to 4 log CFU/ cm2) after processing, was evaluated during storage in vacuum packages at 10 degrees C. Samples were inoculated with a 10-strain composite of Listeria monocytogenes and subsequently immersed (25+/-2 degrees C) for 2 min in 2.5% acetic acid (AA), 2.5% lactic acid (LA), 5% potassium benzoate (PB), or 0.5% Nisaplin (commercial form of nisin, equivalent to 5,000 IU/ml of nisin) solutions, either singly or sequentially (Nisaplin plus AA, Nisaplin plus LA, or Nisaplin plus PB), and then vacuum packaged and stored at 10 degrees C for 48 days. In addition to microbiological analysis, sensory evaluations were performed on uninoculated samples treated with AA, LA, or PB. Initial reductions (day 0) of the pathogen, compared with the controls, on bologna and ham samples treated with AA, LA, or PB ranged from 0.4 to 0.7 log CFU/cm2. Higher (P < 0.05) initial reductions (2.4 to 2.9 log CFU/cm2) were obtained for samples treated with Nisaplin alone and when followed by AA, LA, or PB. L. monocytogenes populations on control bologna and ham samples increased from 3.4 log CFU/cm2 (day 0) to 7.4 and 7.8 log CFU/ cm2, respectively, in 8 days at 10 degrees C. Listericidal effects were observed for all treatments tested, except for Nisaplin applied on its own, during storage at 10 degrees C. The sequential treatment of Nisaplin plus LA reduced L. monocytogenes to undetectable levels in both products at the end of storage. The sequential treatments were also found to inhibit growth of spoilage microorganisms. Sensory evaluations indicated that dipping (2 min) of ham samples in AA (2.5%), LA (2.5%), or PB (5%) led to lower sensory scores. However, since results of this study indicated that these treatments caused extensive listericidal effects, there is possibly a potential to reduce the levels of chemicals applied and still achieve adequate antilisterial activity without major negative effects on product quality.     

The effect of nisin on growth kinetics from activated Bacillus cereus spores in cooked rice and in milk

The growth kinetics of germinated cells from activated spores of Bacillus cereus in cooked white rice and in milk were evaluated at different temperatures for control samples and for samples with 25 microg of nisin per ml added. Nisin was applied in the form of Nisaplin (10(6) IU/g), which contained 25,000 microg of nisin per g. The length of the lag phase for cooked white rice controls was 120 h at 10 degrees C, 8 h at 25 degrees C, and 2.5 h at 33 degrees C. The generation times for cooked rice were 327.7 min at 10 degrees C, 59.0 min at 25 degrees C, and 42.3 min at 33 degrees C; those for milk without nisin were 297.0 min at 20 degrees C, 31.2 min at 30 degrees C, 28.6 min at 35 degrees C, and 33.7 min at 40 degrees C; and those for milk with nisin added were 277.2 min at 20 degrees C, 66.9 min at 30 degrees C, and 66.4 min at 35 degrees C. No development of B. cereus was observed for milk with nisin added at 40 degrees C for 12 h, in which germinated cells decreased by a decimal reduction time (D) of 4.7 h. A temperature of 45 degrees C was shown to be harmful to B. cereus, decreasing the germinated cells in both formulations with D-values of 4.3 to 4.6 h. Similar inhibition of cell growth at 40 degrees C was not observed with lower nisin concentrations.     

Inhibitory combinations of nisin, sodium chloride, and pH on Listeria monocytogenes ATCC 15313 in broth by an experimental design approach

The influence of pH (5.0-8.2), NaCl concentrations (0-6% w/v), and incubation time (0-24 h) on the inhibitory activity of nisin (0-100 I.U./ml) against Listeria monocytogenes (10(3) cfu/ml) was studied using the Doehlert experimental design and was confirmed by kinetic experiments. Predicted values were in agreement with experimental values. Experiments were carried out at 22 degrees C in reconstituted TSB-YE1 broth with or without NaCl. Nisin had an immediate pH-dependent bactericidal effect, which increased with decreasing pH values. In modified TSB-YE1 broth without NaCl, the bactericidal efficacy of nisin (50 I.U./ml) was maximum at pH 6.6, with no L. monocytogenes survivors until 120 h at 22 degrees C. Nisin (50 I.U./ml) action decreased in the presence of NaCl, with a minimal inhibitory effect between 2 and 4%. This partially protective effect was cancelled at higher levels of nisin.     

The influence of nisin on the thermal resistance of Bacillus cereus

Decimal reduction times (D-values) at cooking and autoclaving temperatures (80 to 120 degrees C) of spores of Bacillus cereus ATCC 1479-8 in rice and milk (13% wt/vol) supplemented with nisin (25 microg/ml) were evaluated. The mean D-values at 97.8 degrees C in cooked white rice, phosphate buffer (pH 7.0), and rice water (pH 6.7) were 3.62, 1,99, and 1.34 min, respectively. From 80 to 100 degrees C, the mean reduction in D-values due to the addition of nisin to milk was 40%. The D-value at 110 degrees C was approximately 0.86 min for milk (control) and milk with nisin. The z-values ranged from 7.32 degrees C (phosphate buffer) to 10.37 degrees C (milk control).     

Joint effect of nisin, CO2, and EDTA on the survival of Pseudomonas aeruginosa and Enterococcus faecium in a food model system.

A study on the joint effect of either nisin or Nisaplin, headspace CO2 levels, and EDTA on the survival of Pseudomonas aeruginosa and Enterococcus faecium was carried out in a water-soluble fish muscle extract at 3 degrees C using a second-order rotatable factorial design. E. faecium was completely deactivated by all processing after 2 days of storage. In contrast, P. aeruginosa was much less susceptible to treatments, and cell death was satisfactorily described by two models. Nisin increased cell death, whereas Nisaplin (commercial form of nisin) was not suitable, as it caused undesirable interference, presumably due to its co-compounds. Interactions between Nisaplin or nisin and either EDTA or CO2 were found to be nonstatistically significant. Factors that could account for this unexpected lack of synergism are discussed. However, a statistically significant positive interaction was found between CO2 and EDTA. This finding could allow CO2 levels to be decreased and hence to reduce the main disadvantages of CO2 application, namely, exudation and acidification.     

Influence of nisin on the resistance of Bacillus anthracis sterne spores to heat and hydrostatic pressure

The influence of nisin on the heat and pressure resistance of Bacillus anthracis Sterne spores was examined. The decimal reduction times (D-value) of spores in milk (2% fat) at 80, 85, and 90 degrees C were determined. In the absence of nisin, the D-values were 30.09, 9.30, and 3.86 min, respectively. The D-values of spores heated in the presence of nisin (1 mg/ml) were not significantly different (P = 0.05). However, spores heated in the presence of nisin had a 1- to 2-log reduction in viability, after which the death kinetics became similar to those of spores in the absence of nisin. The z-values all were 11.2 degrees C regardless of the presence or absence of nisin. The pressure sensitivity of B. anthracis Sterne spores in the presence and absence of nisin also was determined. Spores treated with nisin were 10 times more pressure sensitive than were spores subjected to pressure in the absence of nisin under the conditions used in this study.     

Post process control of Listeria monocytogenes on commercial frankfurters formulated with and without antimicrobials and stored at 10 degrees C

The antilisterial effect of postprocess antimicrobial treatments on commercially manufactured frankfurters formulated with and without a 1.5% potassium lactate-0.05% sodium diacetate combination was evaluated. Frankfurters were inoculated (ca. 3 to 4 log CFU/cm2) with 10-strain composite Listeria monocytogenes cultures originating from different sources. The inocula evaluated were cells grown planktonically in tryptic soy broth plus 0.6% yeast extract (30 degrees C, 24 h) or in a smoked sausage homogenate (15 degrees C, 7 days) and cells that had been removed from stainless steel coupons immersed in an inoculated smoked sausage homogenate (15 degrees C, 7 days). Inoculated frankfurters were dipped (2 min, 25 +/- 2 degrees C) in acetic acid (AA; 2.5%), lactic acid (LA; 2.5%), potassium benzoate (PB; 5%), or Nisaplin (commercial form of nisin; 0.5%, equivalent to 5,000 IU/ml of nisin) solutions, or in Nisaplin followed by AA, LA, or PB, and were subsequently vacuum packaged and stored for 48 days at 10 degrees C. In addition to microbiological analyses, sensory evaluations were performed with uninoculated samples that had been treated with AA, LA, or PB for 2 min. Initial L. monocytogenes populations were reduced by 1.0 to 1.8 log CFU/cm2 following treatment with AA, LA, or PB solutions, and treatments that included Nisaplin reduced initial levels by 2.4 to >3.8 log CFU/ cm2. All postprocessing treatments resulted in some inhibition of L. monocytogenes during the initial stages of storage of frankfurters that were not formulated with potassium lactate-sodium diacetate; however, in all cases, significant (P < 0.05) growth occurred by the end of storage. The dipping of products formulated with potassium lactate-sodium diacetate in AA or LA alone--or in Nisaplin followed by AA, LA, or PB-increased lag-phase durations and lowered the maximum specific growth rates of the pathogen. Moreover, depending on the origin of the inoculum, this dipping of products led to listericidal effects. In general, differences in growth kinetics were obtained for the three inocula that were used to contaminate the frankfurters. Possible reasons for these differences include the presence of stress-adapted subpopulations and the inhibition of the growth of the pathogen due to high levels of spoilage microflora. The dipping of frankfurters in AA, LA, or PB did not (P > 0.05) affect the sensory attributes of the product when compared to the control samples. The data generated in this study may be useful to U.S. ready-to-eat meat processors in their efforts to comply with regulatory requirements.     

Combined effects of heat, nisin and acidification on the inactivation of Clostridium sporogenes spores in carrot-alginate particles: from kinetics to process validation

Combined effects of mild temperatures, acidification and nisin on the thermal resistance of Clostridium sporogenes ATCC 11437 spores were assessed. Inoculated carrot-alginate particles were used as a solid-food model for the validation of the spore inactivation during the flow of a solid-liquid food system through the holding tube of an aseptic processing unit. Inactivation kinetics was studied in a water bath with the spores inoculated into carrot-alginate particles and in Sorensen's phosphate buffer. For temperatures of 70-90 degrees C, D-values in the buffer were 24.9-5.7 min, much lower than those evaluated for the particles (115.1-22.2 min). Statistical analyses showed significant synergistic effects of temperature and pH on spore inactivation for both media. Acidification reduced the heat resistance of the spores by reducing the D-values. Nisin was not significantly effective at the lower concentrations (up to 750 IU/g). The combination of 90 degrees C, pH: 4.5 and 500IU/g nisin resulted in a ten-fold decrease of the D-value for spores inoculated in the particles (from 111.1 to 10.6 min). Microbial validation tests were conducted using a pilot-scale aseptic processing unit with a mixture of carrot cubes (10%) and carrier liquid of 2%-carboxymethylcellulose solution (90%). Spore-inoculated carrot-alginate particles (initial counts of 106 CFU/g, obtained after come-up-time pre-heat) with pH 3.5 and 2000 IU/g nisin were processed at 90 degrees C in the aseptic processing unit. Microbial analysis showed no spore survivors in the particles after passing through the holding tube (5.2-6.0 min of residence time). The proposed combination of these hurdles significantly enhanced the spore inactivation rate (D(90)=1.17 min) as compared to that for thermal treatment only (D(90)=19.6 min).     

Inactivation of spores of Bacillus cereus in cheese by high hydrostatic pressure with the addition of nisin or lysozyme

The objective of this work was to study high hydrostatic pressure (HHP) inactivation of spores of Bacillus cereus ATCC 9139 inoculated in model cheeses made of raw milk, together with the effects of the addition of nisin or lysozyme. The concentration of spores in model cheeses was approximately 6-log10 cfu/g of cheese. Cheeses were vacuum packed and stored at 8 degrees C. All samples except controls were submitted to a germination cycle of 60 MPa at 30 degrees C for 210 min, to a vegetative cells destruction cycle of 300 or 400 MPa at 30 degrees C for 15 min, or to both treatments. Bacillus cereus counts were measured 24 h and 15 d after HHP treatment. The combination of both cycles improved the efficiency of the whole treatment. When the second pressure-cycle was of 400 MPa, the highest inactivation (2.4 +/- 0.1 log10 cfu/g) was obtained with the presence of nisin (1.56 mg/L of milk), whereas lysozyme (22.4 mg/L of milk) did not increase sensitivity of the spores to HHP. For nisin (0.05 and 1.56 mg/L of milk), no significant differences were found between counts at 24 h and 15 d after treatment. Considering that mesophilic spore counts usually range from 2.6 to 3.0 log10 cfu/ml in raw milk, HHP at mild temperatures with the addition of nisin may be useful for improving safety and preservation of soft curd cheeses made from raw milk.     

Inactivation of Salmonella enteritidis and Salmonella senftenberg in liquid whole egg using generally recognized as safe additives, ionizing radiation, and heat

The effect of combining irradiation and heat (i.e., irradiation followed by heat [IR-H]) on Salmonella Enteritidis and Salmonella Senftenberg inoculated into liquid whole egg (LWE) with added nisin, EDTA, sorbic acid, carvacrol, or combinations of these GRAS (generally recognized as safe) additives was investigated. Synergistic reductions of Salmonella populations were observed when LWE samples containing GRAS additives were treated by gamma radiation (0.3 and 1.0 kGy), heat (57 and 60 degrees C), or IR-H. The presence of additives reduced the initial radiation Dgamma -values (radiation doses required to eliminate 90% of the viable cells) by 1.2- to 1.5-fold, the thermal decimal reduction times (D,-values) by up to 3.5- and 1.8-fold at 57 and 60 degrees C, respectively, and the thermal D,-values after irradiation treatments by up to 3.4- and 1.5-fold at 57 and 60 degrees C, respectively, for both Salmonella serovars. Of all the additives investigated, nisin at a concentration of 100 IU/ml was the most effective at reducing the heat treatment times needed to obtain a 5-log reduction of Salmonella. Thus, while treatments of 21.6 min at 57 degrees C or of 5 min at 60 degrees C should be applied to achieve a 5-log reduction for Salmonella in LWE, only 5.5 min at 57 degrees C or 2.3 min at 60 degrees C after a 0.3-kGy radiation pretreatment was required when nisin at a concentration of 100 IU/ml was used. The synergistic reduction of Salmonella viability by IR-H treatments in the presence of GRAS additives could enable LWE producers to reduce the temperature or processing time of thermal treatments (current standards are 60'C for 3.5 min in the United States) or to increase the level of Salmonella inactivation.     

Effect of nisin on growth boundaries of Listeria monocytogenes Scott A, at various temperatures, pH and water activities

The effect of nisin on growth boundaries of Listeria monocytogenes Scott A in Tryptone Soy Broth (TSB) under different a(w)s, pH, and temperatures was studied. Growth/no growth turbidity data was modeled using logistic regression. Combinations of various temperatures (5-35 degrees C), pH (4.05-6.70) adjusted with HCl, a(w)s (0.937-0.998) NaCl (0.5-10.5%) and nisin (0-100 IU/ml) were used to monitor the growth/no growth response of L. monocytogenes Scott A for 60 days. The concordance of the logistic regression model was 99.4%, indicating successful data fitting. The minimum pH at which growth was observed was 4.81 at the temperature range of 25-35 degrees C and at a(w) as high as 0.992. Growth was observed at a(w) as low as 0.937, at pH 6.7, at the temperature range of 25-35 degrees C. Increasing nisin concentrations above 25 IU/ml resulted in a more inhibitory environment for L. monocytogenes. Presence of 100 IU/ml resulted in a minimum pH for growth at 5.20, and a minimum a(w) at 0.967 at the temperature range of 25-35 degrees C. It was remarkable that low to medium salt concentrations (2.5-4.5 NaCl% w/v) provided a protective effect against inhibition of L. monocytogenes by nisin. The present study points out the applicability of growth/no growth modeling in order to study any interactions between various factors affecting initiation of growth of micro-organisms, in which its turn helps the understudying of microbe-food ecosystem relations and the development of safer food.     

Combined effect of nisin and carvacrol at different pH and temperature levels on the viability of different strains of Bacillus cereus.

The influence of pH and temperature on the bactericidal action of nisin and carvacrol on vegetative cells of different Bacillus cereus strains was studied. The five strains tested showed significant differences in sensitivity towards nisin, at pH 7.0 and 30 degrees C. Carvacrol concentrations of 0.3 mmol l(-1) had no effect on viability of B. cereus cells. When the same carvacrol concentration was combined with nisin, however, it resulted in a greater loss of viability of cells than when nisin was applied alone. The concentration of carvacrol played an important role on the bactericidal effect of nisin and, therefore, on the synergistic action of both compounds combined. At lower pH values (6.30 and 5.75), nisin was more active against B. cereus cells than at pH 7.0 at 30 degrees C, with a different sensitivity of the strains tested. The combined effect of nisin and carvacrol was found to be significantly different at pH 7.0 and 5.75. When the temperature was 8 degrees C, nisin was significantly less active against B. cereus IFR-NL 94-25 than at 30 degrees C, both at pH 7.0 and 6.30. At 8 degrees C, there was a significant increased effect of nisin at lower pH values. Also at this low temperature, a synergistic effect between nisin and carvacrol on B. cereus cells was observed at the pHs tested. This study indicates the potential of nisin and carvacrol at lower pHs to be used for preservation of minimally processed foods.     

Post-processing application of chemical solutions for control of Listeria monocytogenes, cultured under different conditions, on commercial smoked sausage formulated with and without potassium lactate-sodium diacetate

This study evaluated post-processing chemical solutions for their antilisterial effects on commercial smoked sausage formulated with or without 1.5% potassium lactate plus 0.05% sodium diacetate, and contaminated (approximately 3-4 log cfu/cm(2)) with 10-strain composite Listeria monocytogenes inocula prepared under various conditions. Inoculated samples were left untreated, or were immersed (2 min, 25 +/- 2 degrees C) in solutions of acetic acid (2.5%), lactic acid (2.5%), potassium benzoate (5%) or Nisaplin (0.5%, equivalent to 5000 IU/ml of nisin) alone, and in sequence (Nisaplin followed by acetic acid, lactic acid or potassium benzoate), before vacuum packaging and storage at 10 degrees C (48 days). Acetic acid, lactic acid or potassium benzoate applied alone reduced initial L. monocytogenes populations by 0.4-1.5 log cfu/cm(2), while treatments including Nisaplin caused reductions of 2.1-3.3 log cfu/cm(2). L. monocytogenes on untreated sausage formulated with antimicrobials had a lag phase duration of 10.2 days and maximum specific growth rate (mu(max)) of 0.089 per day, compared to no lag phase and mu(max) of 0.300 per day for L. monocytogenes on untreated product that did not contain antimicrobials in the formulation. The immersion treatments inhibited growth of the pathogen for 4.9-14.8 days on sausage formulated without potassium lactate-sodium diacetate; however, in all cases significant (P < 0.05) growth occurred by the end of storage. The antilisterial activity of chemical solutions was greatly enhanced when applied to product formulated with antimicrobials; growth was completely inhibited on sausage treated with acetic or lactic acid alone, and in sequence with Nisaplin. In general, habituation (15 degrees C, 7 days) of L. monocytogenes cells, planktonically or as attached cells to stainless-steel coupons in sausage homogenate prior to contamination of product, resulted in shorter lag phase durations compared with cells cultivated planktonically in a broth medium. Furthermore, when present, high levels of spoilage flora were found to suppress growth of the pathogen. Findings of this study could be useful to US meat processors in their efforts to select required regulatory alternatives for control of post-processing contamination in meat products.     

Further Studies on the Antibotulinal Effectiveness of Nisin in Acidic Media

The antibotulinal effectiveness of nisin in TPYG broth was increased somewhat by lowering the pH to pH 5.5. The ability of nisin to inhibit the outgrowth of strain 56A spores was markedly increase at pH 5.5 by comparison to its effectiveness at higher pHs observed in previous studies. The increased effectiveness of nisin at pH 5.5 was less notable for the strain 69A, 113B, and 213B spores. The nisin sensitivity of the type E spores was essentially unchanged from that observed in earlier studies at higher pHs. At pH 6, nisin levels of 5000 I.U./ml were insufficient to prevent spore outgrowth by C. botulinum in cooked meat medium. Comparatively, much lower levels of nisin were effective in preventing botulinal outgrowth in TPYG broth at pH 6. The decreased effectiveness of nisin in cooked meat medium may be due to the binding of nisin to meat particles, and this binding is apparently not affected by lowering the pH to pH 6.0.     

Heat resistance of Yersinia enterocolitica grown at different temperatures and heated in different media.

In the range of 4-20 degrees C, growth temperature did not influence the heat resistance at 54-66 degrees C for Yersinia enterocolitica at pH 7 in citrate phosphate buffer. However, when cells were grown at 37 degrees C. the D62 increased from 0.044 to 0.17 min. This increase was constant at all heating temperatures tested (z = 5.7-5.8). Growth temperature did not influence the proportion of heat-damaged cells after a heat treatment, as measured by their response to a 2% of sodium chloride added to the recovery medium. The sensitivity of heat treated cells to nisin or lysozyme depended on growth temperature: Whereas the number of cells grown at 4 degrees C surviving heat treatment was the same regardless of the presence of 100 IU/ml of nisin or 100 microg/ml of lysozyme in the recovery medium, that of cells grown at 37 degrees C was, in these media, lower. The pH of maximum heat resistance in citrate phosphate buffer was pH 7 for cells grown at 37 degrees C, but pH 5 for those grown at 4 degrees C. In both suspensions the magnitude of the effect of pH on heat resistance was constant at all heating temperatures. For cells grown at 4 degrees C the heat resistance at 54-66 degrees C, in skimmed milk or pH 7 buffer, was the same. For cells grown at 37 degrees C this also applied for heat treatment at 66 degrees C but at 56 degrees C the heat resistance in skimmed milk was higher.     

Effect of Nisin on the Outgrowth of Clostridium botulinum Spores

Nisin, an antibiotic produced by certain strains of Streptococcus lactis, is effective in preventing the outgrowth of Clostridium botulinum spores. Type A C. botulinum spores were the most resistant to the inhibitory action of nisin requiring 1000-2000 I.U. of nisin/ml for a 50% inhibition of outgrowth on TPYG agar plates. Type E spores were more sensitive requiring only 50-100 I.U./ml for 50% inhibition of outgrowth on TPYG agar plates. Type B spores displayed an intermediate level of sensitivity requiring 500-1000 I.U. of nisin/ml for 50% inhibition of outgrowth on TPYG agar plates. Similar levels of nisin were necessary to prevent spore outgrowth in TPYG broth and BHI broth over a 7-day incubation period. With prolonged incubation periods of up to 65 days in TPYG broth, spore outgrowth was observed sporadically at higher nisin levels with the type A and B spores which may indicate some decomposition of nisin with storage. Nisin levels of 5000 I.U./ml for the type A spores and 2000 I.U./ml for the type B spores and the Minnesota E spores were insufficient to prevent spore outgrowth by C. botulinurn in cooked meat medium. For the Beluga E spores, a nisin level of 2000 I.U./ml was necessary to prevent spore outgrowth in cooked meat medium. The need for higher levels of nisin in cooked medium to prevent spore outgrowth may be due to the binding of the nisin by meat particles.     

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.     

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.