Lilian Nilsson

Inhibition of Listeria monocytogenes on cold-smoked salmon by nisin and carbon dioxide atmosphere

The bacteriostatic and bacteriocidal effect of nisin in combination with carbon dioxide, NaCl and low temperature on the survival of Listeria monocytogenes was investigated in in vitro model studies and in trials with cold-smoked salmon. Addition of nisin caused various degrees of inhibition and sometimes death of L. monocytogenes in model experiments performed at 10 degrees C. The antilisterial effect of nisin was improved in the presence of 100% CO2 and increasing NaCl concentrations (0.5 to 5.0% w/v). Minimal bactericidal concentrations (MBC) of nisin varied from 30 to more than 500 IU/ml. The most pronounced effect of nisin was found when 10(2) cfu/ml was grown in media with 5.0% NaCl and incubated in CO2 atmosphere (MBC = 30 IU/ml). The bactericidal effect of nisin was reduced in air and vacuum, and did not increase systematically with increasing NaCl concentrations. In general, nisin concentration < or = 50 IU/ml resulted in the survival and growth of L. monocytogenes in all combinations with other preservatives (NaCl, CO2). Addition of nisin (500 or 1000 IU/g) to cold-smoked salmon inoculated with L. monocytogenes and stored at 5 degrees C delayed, but did not prevent growth of L. monocytogenes in vacuum-packs. Numbers of L. monocytogenes increased to 10(8) cfu/g in vacuum packed cold-smoked salmon in 8 days, whereas CO2 packing of cold-smoked salmon resulted in an 8-day lag phase of L. monocytogenes, with numbers eventually reaching 10(6) cfu/g in 27 days. Addition of nisin to CO2 packed cold-smoked salmon resulted in a 1 to 2 log reduction of L. monocytogenes followed by a lag phase of 8 and 20 days in salmon with 500 and 1000 IU nisin/g, respectively. The levels of L. monocytogenes remained below 10(3) cfu/g during 27 days of storage at both concentrations of nisin.

Growth control of Listeria monocytogenes on cold-smoked salmon using a competitive lactic acid bacteria flora.

A Lactobacillus sake strain LKE5 and four strains of Carnobacterium piscicola were evaluated as biopreservation cultures to control the growth of Listeria monocytogenes on vacuum-packed, cold-smoked salmon stored at 5 degrees C. All five strains were antilisterial as live cultures in an agar diffusion assay. Cell-free supernatants of two strains of C. piscicola and L. sake LKE5 were also antilisterial because of the production of bacteriocins. The presence of high cell numbers of strains of C. piscicola had no influence on the sensory quality of cold-smoked salmon stored at 5 degrees C, but L. sake LKE5 caused strong sulfurous off-flavors and was rejected as a culture for biopreservation of cold-smoked salmon. A bacteriocin-producing strain of C. piscicola (A9b) initially caused a 7-day lag phase of L. monocytogenes, followed by a reduction in numbers of L. monocytogenes from 10(3) CFU/ml to below 10 CFU/ml after 32 days of incubation, coinciding with the detection of antilisterial compounds. The presence of a nonbacteriocin-producing strain of C. piscicola (A10a) prevented the growth of L. monocytogenes during the 32-day incubation. The growth of L. monocytogenes was strongly repressed on cold-smoked salmon in the presence of C. piscicola A9b and A 10a, respectively. The initial cell numbers of L. monocytogenes that were found on Oxford plates incubated at 25 degrees C reached low maximum cell counts of 10(4) and 2 x 10(3) after 14 and 20 days of storage in mixed culture with C. piscicola A9b and A10a.

Carbon Dioxide and Nisin Act Synergistically on Listeria monocytogenes

ABSTRACT This paper examines the synergistic action of carbon dioxide and nisin on Listeria monocytogenes Scott A wild-type and nisin-resistant (Nisr) cells grown in broth at 4°C. Carbon dioxide extended the lag phase and decreased the specific growth rate of both strains, but to a greater degree in the Nisrcells. Wild-type cells grown in 100% CO2 were two to five times longer than cells grown in air. Nisin (2.5 μg/ml) did not decrease the viability of Nisr cells but for wild-type cells caused an immediate 2-log reduction of viability when they were grown in air and a 4-log reduction when they were grown in 100% CO2. There was a quantifiable synergistic action between nisin and CO2 in the wild-type strain. The MIC of nisin for the wild-type strain grown in the presence of 2.5 μg of nisin per ml increased from 3.1 to 12.5 μg/ml over 35 days, but this increase was markedly delayed for cultures in CO2. This synergism between nisin and CO2 was examined mechanistically by following the leakage of carboxyfluorescein (CF) from listerial liposomes. Carbon dioxide enhanced nisin-induced CF leakage, indicating that the synergistic action of CO2 and nisin occurs at the cytoplasmic membrane. Liposomes made from cells grown in a CO2 atmosphere were even more sensitive to nisin action. Liposomes made from cells grown at 4°C were dramatically more nisin sensitive than were liposomes derived from cells grown at 30°C. Cells grown in the presence of 100% CO2 and those grown at 4°C had a greater proportion of short-chain fatty acids. The synergistic action of nisin and CO2 is consistent with a model where membrane fluidity plays a role in the efficiency of nisin action.

The contribution of bacteriocin to inhibition of Listeria monocytogenes by Carnobacterium piscicola strains in cold-smoked salmon systems

Aims:  To study the importance of bacteriocin production for the antilisterial effect of a bacteriocinogenic Carnobacterium piscicola strain A9b on growth of Listeria monocytogenes in broth and cold‐smoked salmon systems.

Growth inhibition of Listeria monocytogenes by a nonbacteriocinogenic Carnobacterium piscicola

Aims:  This study elucidates the mechanisms by which a nonbacteriocinogenic Carnobacterium piscicola inhibits growth of Listeria monocytogenes.

Factors affecting production of an antilisterial bacteriocin by Carnobacterium piscicola strain A9b in laboratory media and model fish systems.

Aims: To investigate factors influencing bacteriocin production and bacteriocin stability of the bioprotective culture Carnobacterium piscicola strain A9b.

Role of acetate in production of an autoinducible Class IIa Bacteriocin in Carnobacterium piscicola A9b

ABSTRACT Carnobacterium piscicola strain A9b isolated from cold smoked salmon inhibits growth of the food-borne pathogen Listeria monocytogenes partly due to the production of a proteinaceous compound (L. Nilsson, L. Gram, and H. H. Huss. J. Food Prot. 62:336-342, 1999). The purpose of the present study was to purify the compound and describe factors affecting its production, with particular emphasis on food-relevant factors. Amino acid sequencing showed that the compound is a class IIa bacteriocin with an N-terminal amino acid sequence identical to that of carnobacteriocin B2. The production of the bacteriocin was autoinducible, and the threshold level for induction was 9.6 × 10−10 M. We also report, for the first time, that acetate acts as an induction factor, with a threshold concentration of 0.3 to 12 mM. Acetate could not act as an inducer during the late exponential phase of C. piscicola A9b. The induction of bacteriocin production showed a dose-dependent relationship at acetate concentrations of up to 10 to 20 mM (depending on the growth medium) and at a concentration of 1.9 × 10−8 M for the bacteriocin itself; a saturation level of bacteriocin specific activity was reached at these concentrations of induction factors. The combined use of both inducers did not enhance the saturation level of bacteriocin production compared to that seen with the use of each inducer alone. Increasing NaCl and glucose concentrations negatively influenced the efficiency of acetate as an induction factor. Based on the results, carnobacteriocin B2 was used as an induction factor to manipulate the production of bacteriocin in cold smoked salmon juice and thus improve the ability to inhibit L. monocytogenes.