Evaluation of nisin-coated cellulose casings for the control of Listeria monocytogenes inoculated onto the surface of commercially prepared frankfurters

Commercially prepared frankfurters were formulated with and without approximately 1.4% potassium lactate and 0.1% sodium diacetate and were subsequently processed in cellulose casings coated with and without nisin (approximately 50,000 IU per square inch of internal surface area) to control the outgrowth of Listeria monocytogenes during refrigerated storage. The frankfurters were inoculated with approximately 5 log CFU per package of a five-strain mixture of L. monocytogenes and then vacuum sealed before being stored at 4 degrees C for 60 to 90 days. Surviving organisms were recovered and enumerated by rinsing each package with 18 ml of sterile 0.1% peptone water and plating onto MOX selective agar. The data for each of two trials were averaged. In packages that contained frankfurters formulated with potassium lactate and sodium diacetate and prepared in nisin-coated casings, L. monocytogenes levels decreased by 1.15 log CFU per package after 90 days of storage. L. monocytogenes levels decreased by 0.95 log CFU per package in frankfurters that were prepared in casings that were not coated with nisin. In packages of frankfurters that were formulated without potassium lactate and sodium diacetate and prepared in nisin-coated casings, L. monocytogenes levels decreased by 0.88 log CFU per package after 15 days of storage but then increased appreciably thereafter over a 60-day period of refrigerated storage. There was also an appreciable increase in pathogen numbers during 60 days of storage in otherwise similar frankfurters formulated without potassium lactate and sodium diacetate prepared in casings that were not coated with nisin. These data confirm that potassium lactate and sodium diacetate display listeriostatic activity as an ingredient of commercial frankfurters. These data also establish that cellulose casings coated with nisin display only moderate antilisterial activity in vacuum-sealed packages of commercially prepared frankfurters 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 PROPERTIES OF NISIN-COATED POLYMERIC FILMS AS INFLUENCED BY FILM TYPE AND COATING CONDITIONS

Polymeric films were coated with nisin, an antimicrobial peptide, for potential application in shelf-life extension of food. Five commercial packaging films with varying surface energies were compared. The highest antimicrobial activity was observed on the most hydrophobic nisin-coated films (surface energy ± 32 dyne/cm). The binding affinity of nisin to the surface of ethylene acrylic acid copolymer (surface energy = 32 dyne/cm) was investigated. Films were coated in nisin solutions under varying conditions including nisin concentration, contact time, solution's volume to film area ratio, pH of coating solution and temperature during coating. Stronger nisin activity was detected on films as the coating time increased from 30 s to 2 h. Higher nisin coating concentration also promoted the binding of nisin to films with saturation level observed at 250 μg/mL. Films coated with nisin using a higher solution to film area ratio (0.84 compared to 0.14 and 0.28 mL/cm²) also exhibited higher nisin activity.     

Effectiveness of chitosan-coated plastic films incorporating antimicrobials in inhibition of Listeria monocytogenes on cold-smoked salmon

The objective of this study was to evaluate the efficacy of chitosan-coated plastic films incorporating five Generally Recognized as Safe (GRAS) antimicrobials (nisin, sodium lactate (SL), sodium diacetate (SD), potassium sorbate (PS) and sodium benzoate (SB)) against Listeria monocytogenes on cold-smoked salmon. Salmon samples were surface-inoculated with a five-strain cocktail of L. monocytogenes and packaged in chitosan-coated plastic films containing 500 IU/cm(2) of nisin, 9 mg/cm(2) of SL, 0.5 mg/cm(2) of SD, 0.6 mg/cm(2) of PS, or 0.2 mg/cm(2) of SB, and stored at room temperature (ca. 20 degrees C) for 10 days. The film incorporating SL was the most effective, completely inhibiting the growth of L. monocytogenes during 10 days of storage. L. monocytogenes in samples packaged in the other four antimicrobial films grew, but the increase in counts was lower than the control. The antilisterial efficacy of films containing lower concentrations of SL (2.3 mg/cm(2) and 4.5 mg/cm(2)) and binary combinations SL, PS, SD, SB and nisin were subsequently evaluated. Among all the treatments, chitosan-coated plastic films with 4.5 mg/cm(2) SL, 4.5 mg/cm(2) SL-0.6 mg/cm(2) PS and 2.3 mg/cm(2) SL-500 IU/cm(2) nisin were the most effective. These three most effective antimicrobial films were then tested at refrigerated temperature. They completely inhibited the growth of L. monocytogenes on smoked salmon for at least 6 weeks. Chitosan-coated plastic films containing 4.5 mg/cm(2) SL can potentially assist the smoked-salmon processing industry in their efforts to control L. monocytogenes.     

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.     

Control of Listeria monocytogenes on ham steaks by antimicrobials incorporated into chitosan-coated plastic films

Contamination of ready-to-eat (RTE) meat products such as ham steaks with Listeria monocytogenes has been a concern for the meat processing industry. The objective of this study was to evaluate the antilisterial efficacy of chitosan-coated plastic films alone or incorporating five generally recognized as safe (GRAS) antimicrobials. Effect of chitosan-coated plastic film on the growth of L. monocytogenes was first investigated in an aqueous system of culture medium broth and chitosan-coated films were able to inhibit the growth of L. monocytogenes in a concentration-dependent manner. However, chitosan-coated plastic films were not able to control the growth of L. monocytogenes on ham steaks. Therefore, five GRAS antimicrobials were subsequently incorporated into chitosan-coated plastic films to enhance their antilisterial effectiveness. Ham steaks were surface-inoculated with a five-strain cocktail of L. monocytogenes and then packaged in chitosan-coated plastic films containing 500 IU/cm(2) of nisin, 0.01 g/cm(2) of sodium lactate (SL), 0.0025 g/cm(2) of sodium diacetate, 0.003 g/cm(2) of potassium sorbate (PB), or 0.001 g/cm(2) of sodium benzoate (SB). The samples were stored at room temperature (ca. 20 degrees C) for 10 days. Incorporating antimicrobials into chitosan-coated plastic films slowed down or inhibited the growth of L. monocytogenes. The chitosan-coated plastic film containing SL was the most effective antimicrobial film and its efficacy against L. monocytogenes on ham steaks was evaluated during 12-week storage at 4 degrees C. The film showed excellent long-term antilisterial effect with the counts of L. monocytogenes being slightly lower than the initial inoculum. Chitosan-coated plastic films containing 0.001 g/cm(2) of SL have a potential to be used on ham steaks to control L. monocytogenes.     

Anti-listerial activity of a polymeric film coated with hybrid coatings doped with Enterocin 416K1 for use as bioactive food packaging

In this study, Enterocin 416K1, a bacteriocin produced by Enterococcus casseliflavus IM 416K1, was entrapped in an organic-inorganic hybrid coating applied to a LDPE (low-density polyethylene) film for its potential use in the active food packaging field. The antibacterial activity of the coated film was evaluated against Listeria monocytogenes NCTC 10888 by qualitative modified agar diffusion assay, quantitative determination in listeria saline solution suspension and direct contact with artificially contaminated food samples (frankfurters and fresh cheeses) stored at room and refrigeration temperatures. All investigations demonstrated that enterocin-activated coatings have a good anti-listeria activity. Qualitative tests showed a clear zone of inhibition in the indicator lawn in contact with and around the coated film. During the quantitative antibacterial evaluation the L. monocytogenes viable counts decreased to 1.5 log units compared to the control. The inhibitory capability was confirmed also in food-contact assays. In all food samples packed with coated films we observed a significant decrease in L. monocytogenes viable counts in the first 24 h compared to the control. This difference was generally maintained up to the seventh day and then decreased, with the exception of the cheese samples stored at refrigeration temperature.     

Release of Nisin from Methylcellulose-Hydroxypropyl Methylcellulose Film Formed on Low-density Polyethylene Film

ABSTRACT: Low-density polyethylene film was coated with a solution containing a high-molecular-weight or low-molecular-weight methylcellulose and hydroxypropyl methylcellulose. Films contained 10000, 7500, 5000, 2500, or 0 IU/cm2 nisin. Film samples were placed into peptone water, and 10-μL samples were removed and placed onto spiral plated lawns of Listeria monocytogenes . Zones of inhibition were measured using a caliper. Films containing 5000, 7500, and 10000 IU/cm2 nisin inhibited L. monocytogenes after 30 min; films with 7500 and 10000 IU/cm2 nisin inhibited L. monocytogenes after 60 min and 8 h, respectively. No zones of inhibition were observed after 24 h and 4 d for all films. After 8 d, zones of inhibition were observed for films with all levels of nisin except 2500 IU/cm2. Films with 0 and 2500 IU/ cm2 nisin did not produce zones of inhibition throughout the study. Molecular weight of the cellulose-based carrier had no effect on inhibition of L. monocytogenes. A standard curve of inhibition was developed using solutions of 10000, 7500, 5000, 2500, or 0 IU/cm2 nisin applied directly to lawns of L. monocytogenes . The amount of inhibition using a direct application of a solution was 25% to 50% more effective for inhibition of L. monocytogenes compared with the coated film samples. Overall, the coated film samples were effective for inhibition of L. monocytogenes , particularly when 7500 and 10000 IU/cm2 nisin were used, but the release of nisin was not controlled and did not provide consistent inhibition throughout the 8-d study.     

Biodegradable polylactic acid polymer with nisin for use in antimicrobial food packaging

ABSTRACT:  Biodegradable polylactic acid (PLA) polymer was evaluated for its application as a material for antimicrobial food packaging. PLA films were incorporated with nisin to for control of foodborne pathogens. Antimicrobial activity of PLA/nisin films against Listeria monocytogenes , Escherichia coli O157:H7, and Salmonella Enteritidis were evaluated in culture media and liquid foods (orange juice and liquid egg white). Scanned electron micrograph and confocal laser microscopy revealed that nisin particles were evenly distributed in PLA polymer matrix on the surface and inside of the PLA/nisin films. PLA/nisin significantly inhibited growth of L . monocytogenes in culture medium and liquid egg white. The greatest inhibition occurred at 24 h when the cell counts of L. monocytogenes in the PLA/nisin samples were 4.5 log CFU/mL less than the controls. PLA/nisin reduced the cell population of E. coli O157:H7 in orange juice from 7.5 to 3.5 log at 72 h whereas the control remained at about 6 log CFU/mL. PLA/nisin treatment resulted in a 2 log reduction of S. Enteritidis in liquid egg white at 24 °C. After 21 d at 4 °C the S. Enteritidis population from PLA/nisin treated liquid egg white (3.5 log CFU/mL) was significantly less than the control (6.8 log CFU/mL). E. coli O157:H7 in orange juice was more sensitive to PLA/nisin treatments than in culture medium. The results of this research demonstrated the retention of nisin activity when incorporated into the PLA polymer and its antimicrobial effectiveness against foodborne pathogens. The combination of a biopolymer and natural bacteriocin has potential for use in antimicrobial food packaging.     

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.     

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.     

Antimicrobial activity of lactoferrin against foodborne pathogenic bacteria incorporated into edible chitosan film

The objectives of this research were to develop and characterize edible chitosan film containing lactoferrin as a natural antimicrobial agent, and to investigate the combination effects of lactoferrin with lysozyme in chitosan film against the growth of Escherichia coli O157:H7 and Listeria monocytogenes. Chitosan films containing lactoferrin, lysozyme, or nisin were fabricated, and the antimicrobial concentrations were 0.5, 1, or 2 mg in a circular disc of chitosan film. Three concentrations of lactoferrin or EDTA (0.28, 0.56, or 1.12 mg per disc) were also incorporated into the chitosan film containing lysozyme to investigate the combination effects of lactoferrin. The water barrier properties of the chitosan films containing lactoferrin were characterized. The antimicrobial activities against E. coli O157:H7 and L. monocytogenes were determined using the agar diffusion assay and cell count assay. The chitosan films containing lactoferrin less than 1 mg per disc did not alter the water vapor permeability of the chitosan film. Although the film containing lysozyme exhibited significant antimicrobial activity, the incorporation of lactoferrin alone into chitosan film did not exhibit significant antimicrobial activity against both E. coli O157:H7 and L. monocytogenes. However, the combination of lactoferrin with lysozyme-containing chitosan film significantly decreased the growth of E. coli O157:H7, exhibiting a comparable effect to that of the combination of EDTA with lysozyme (P < 0.05). Furthermore, the combination of lactoferrin with lysozyme in chitosan film exhibited greater reduction in the growth of L. monocytogenes than did the combination EDTA with lysozyme, resulting in an approximate 3-log reduction.     

Enhancing Physical Properties and Antimicrobial Activity of Konjac Glucomannan Edible Films by Incorporating Chitosan and Nisin

ABSTRACT: The antimicrobial effect of konjac glucomannan (KGM) edible ûlm incorporating chitosan (CHI) and nisin at various ratios or concentrations was studied. This activity was tested against pathogenic bacteria, namely, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes , and Bacillus cereus. Mechanical and physical properties were determined, and the results indicated that the blend film KC2 (mixing ratio KGM 80/ CHI 20) showed the maximum tensile strength (102.8 ± 3.8 MPa) and good transparency, water solubility, and water vapor transmission ratio. Differential scanning calorimetry (DSC) and Fourier transform infrared (FTIR) spectroscopy were used to characterize the structural change of the blend films; and the results showed that strong intermolecular hydrogen bonds occurred between CHI and KGM. Incorporation of nisin at 463IU per disk of film for the selected KC2 was found to have antimicrobial activity against S. aureus, L. monocytogenes , and B. cereus. The mean value of inhibition zone diameter of the CHI-N series and the KC2-N series were higher than the KGM-N series at each corresponding concentration and with significant difference ( P < 0.05), however, there was no significant difference in the antimicrobial effect between CHI and KC2 incorporating nisin. At all these levels, the blend ûlm KC2-nisin had a satisfactory appearance, mechanical and physical properties, and antimicrobial activity. Therefore, it could be considered as a potential "active" packaging material.     

Effects of combinations of lactoperoxidase system and nisin on the behaviour of Listeria monocytogenes ATCC 15313 in skim milk

Individual or combined effects of nisin (100 or 200 IU/ml) and the lactoperoxidase system (LPS) were analysed against 1 x 10(4) cfu/ml Listeria monocytogenes ATCC 15313 cells in skim milk, at 25 degrees C for 15 days. Nisin induced an immediate bactericidal effect and LPS a 48 h bacteriostatic phase which in both cases was followed by re-growth of L. monocytogenes. LPS and nisin added together at t0 showed a synergistic and lasting bactericidal effect which after 8 days and until 15 days resulted in no detectable cells in 1 ml of milk. When LPS was added to cells already in contact with 100 or 200 IU/ml nisin for a period of 4 h, the inhibitory activity was enhanced with no L. monocytogenes detectable after 72 or 48 h, respectively, and until 15 days. When LPS was added after 12 h, the nisin bactericidal phase was followed by re-growth. When nisin, 100 or 200 UI/ml, was added to cells already in contact with LPS over 24 h, L. monocytogenes was not detectable after 196 and 244 h, respectively, without any re-growth. For nisin addition after 72 h, cell counts were 8 log10 cycles lower than in the control milk after 196 h, but population levels were similar to the control within 15 days. The best combination to inhibit L. monocytogenes ATCC 15313 was nisin present at t0 followed by the LPS addition 4 h later, when the maximum inhibitory effect of nisin was reached.     

Control of Listeria monocytogenes and Salmonella anatum on the surface of smoked salmon coated with calcium alginate coating containing oyster lysozyme and nisin

ABSTRACT:  This study investigated the antimicrobial effect of oyster lysozyme with or without nisin added to calcium alginate (CaAlg) coated on the surface of smoked salmon against Listeria monocytogenes and Salmonella anatum . L. monocytogenes or S. anatum inoculated smoked salmon samples (1 g) were dipped into CaAlg with either oyster lysozyme (OysL) or hen egg white lysozyme (HEWL), with or without added nisin (N), then stored at 4 °C for 35 d. Our results indicated that the effectiveness of oyster lysozyme or hen egg white lysozyme was enhanced when added to calcium alginate coatings. After 35 d at 4 °C the growth of L. monocytogenes and S. anatum was suppressed in the range of 2.2 to 2.8 log CFU/g with CaAlgNOysL or CaAlgNHEWL coatings compared to the control nontreated samples. There was no significant difference between oyster lysozyme and hen egg white lysozyme treatments against L. monocytogenes or S. anatum inoculated on the surface of salmon. Calcium alginate coatings containing lysozyme with nisin or without could be used to reduce the growth of L. monocytogenes and S. anatum on the surface of ready-to-eat smoked salmon at refrigerated temperatures.     

Antimicrobial food packaging film based on the release of LAE from EVOH

The aim of this work was to develop antimicrobial films for active packaging applications containing the natural antimicrobial compound LAE (lauramide arginine ethyl ester) in EVOH copolymers with different mol % ethylene contents (i.e. EVOH-29 and EVOH-44). EVOH-29 and EVOH-44 films were made by casting and incorporating 0.25%, 1%, 5%, and 10% LAE in the film forming solution (w/w with respect to polymer weight). Previously, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of LAE against Listeria monocytogenes, Escherichia coli, and Salmonella enterica were determined by a microdilution assay. The antimicrobial activity of the resulting films was tested in vitro against these microorganisms in liquid culture media. The activity of the films was also evaluated over time. The results showed that films containing 5% and 10% LAE produced total growth inhibition and viable counts decreased with 0.25% and 1% LAE. Finally, the effectiveness of the films was tested by applying them to an infant formula milk inoculated with L. monocytogenes and S. enterica and stored for 6 days at 4°C. The application of films with LAE to infant formula milk inoculated with L. monocytogenes reduced at the end of storage period about 4 log in case of 10% LAE and with S. enterica reduced 3.74 log and 3.95 log with EVOH 29 5% and 10%, respectively, and EVOH-44 5% and 10% LAE reduced 1 log and 3.27 log, respectively, at the end of storage. The antimicrobial capacity of EVOH-29 films was greater than that of EVOH-44 films in all the cases tested. In general, the films were more effective in inhibiting the growth of L. monocytogenes than S. enterica, this inhibition being more acute at the end of the storage time.     

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.     

Enhancement of nisin, lysozyme, and monolaurin antimicrobial activities by ethylenediaminetetraacetic acid and lactoferrin

A microtiter plate assay was employed to systematically assess the interaction between ethylenediaminetetraacetic acid (EDTA) or lactoferrin and nisin, lysozyme, or monolaurin against strains of Listeria monocytogenes, Escherichia coli, Salmonella enteritidis, and Pseudomonas fluorescens. Low levels of EDTA acted synergistically with nisin and lysozyme against L. monocytogenes but EDTA and monolaurin interacted additively against this microorganism. EDTA synergistically enhanced the activity of nisin, monolaurin, and lysozyme in tryptic soy broth (TSB) against two enterohemorrhagic E. coli strains. In addition, various combinations of nisin, lysozyme, and monolaurin with EDTA were bactericidal to some gram-negative bacteria whereas none of the antimicrobials alone were bactericidal. Lactoferrin alone (2000 microg ml(-1)) did not inhibit any of the bacterial strains, but did enhance nisin activity against both L. monocytogenes strains. Lactoferrin in combination with monolaurin inhibited growth of E. coli O157:H7 but not E. coli O104:H21. While lactoferrin combined with nisin or monolaurin did not completely inhibit growth of the gram-negative bacteria, there was some growth inhibition. All combinations of EDTA or lactoferrin with antimicrobials were less effective in 2% fat UHT milk than in TSB. S. enteritidis and P. fluorescens strains were consistently more resistant to antimicrobial combinations. Resistance may be due to differences in the outer membrane and/or LPS structure.     

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.     

Efficacy of nisin in combination with protective cultures against Listeria monocytogenes Scott A in tofu.

Nisin can be used as a biopreservative to control growth of Listeria monocytogenes in various minimally processed foods. Tofu is an example of a non-fermented soybean product, which may allow growth of Listeria at refrigeration temperatures and in which nisin may be applied to prevent multiplication of Listeria. The efficacy of nisin against Listeria may be compromised by the emergence of spontaneous nisin-resistant mutants. Exposure of L. monocytogenes Scott A to nisin in a culture medium or in a food product results in an initial reduction of Listeria population which is followed by regrowth of survivors to nisin during further incubation. In vitro studies using Standard I Nutrient broth showed that Enterococcus faecium BFE 900-6a and Lactobacillus sakei Lb 706-1a used as protective cultures in combination with nisin were able to suppress proliferation of Listeria cells not killed by nisin at 10 degrees C. Growth and bacteriocin production of these two strains and a third protective culture, Lactococcus lactis BFE 902 was also observed in soymilk and tofu at 10 degrees C. Inoculation studies with tofu prepared with nisin and protective cultures showed that lower amounts of nisin are required for an effective inhibition of L. monocytogenes Scott A when either E. faecium BFE 900-6a or Lc. lactis BFE 902 are used in addition. The combination of nisin with these bacteriocinogenic lactic acid bacteria (LAB) resulted in a complete suppression of listerial growth in homemade tofu stored at 10 degrees C for 1 week. Lb. sakei Lb 706-1a was less effective and did not prevent a slight increase of L. monocytogenes Scott A numbers during storage.