Interaction of water activity and bicarbonate salts in the inhibition of growth and mycotoxin production by Fusarium and Aspergillus species of importance to corn.

The combined effects of water activity (a(w)) and ammonium/sodium bicarbonate on growth and mycotoxin production in corn by Fusarium and Aspergillus species were investigated. Interaction was observed between the salts and a(w) on the colony growth rates and lag phase durations of all isolates. Growth stimulation at low salt levels was observed only for the Fusarium isolates as the fastest growth of F. verticillioides and F. proliferatum occurred at levels of 0.1-0.2 and 0.5% ammonium and sodium bicarbonate, respectively. Although the complete inhibition of the growth of the Fusarium and Aspergillus isolates investigated took place at a level of 1% ammonium bicarbonate as much as 4% sodium bicarbonate failed to completely inhibit the growth of the Aspergillus isolates. Increase in concentration of either salt generally resulted in large reductions of both fumonisin B(1) and aflatoxin B(1) production. According to the sensorial analysis performed, corn treated with up to 1% ammonium bicarbonate was still acceptable for consumption, whereas corn treated with at least 2% sodium bicarbonate was determined to be sensorially unsuitable. Ammonium bicarbonate can be concluded to be more suitable for protecting stored corn from fungal contamination as it was capable of completely inhibiting both growth and mycotoxin production of the Fusarium and Aspergillus isolates of most importance to corn at levels that were still sensorially acceptable. Therefore ammonium bicarbonate could possibly be applied as a cheap and easy to apply treatment for use in resource limited developing countries.

Predictive modelling of the individual and combined effect of water activity and temperature on the radial growth of Fusarium verticilliodes and F. proliferatum on corn.

The major objective of this study was to develop validated models to describe the effect of a(w) and temperature on the radial growth on corn of the two major fumonisin producing Fusaria, namely Fusarium verticilliodes and F. proliferatum. The growth of these two isolates on corn was therefore studied at water activities between 0.810-0.985 and temperatures between 15 and 30 degrees C. Minimum a(w) for growth was 0.869 and 0.854 for F. verticilliodes and F. proliferatum, respectively. No growth took place at a(w) values equal to 0.831 and 0.838 for F. verticilliodes and F. proliferatum, respectively. The colony growth rates, g (mm d(-1)) were determined by fitting a flexible growth model describing the change in colony diameter (mm) with respect to time (days). Secondary models, relating the colony growth rate with a(w) or a(w) and temperature were developed. A third order polynomial equation and the linear Arrhenius-Davey model were used to describe the combined effect of temperature and a(w) on g. The combined modelling approaches, predicting g (mm d(-1)) at any a(w) and/or temperature were validated on independently collected data. All models proved to be good predictors of the growth rates of both isolates on maize within the experimental conditions. The third order polynomial equation had bias factors of 1.042 and 1.054 and accuracy factors of 1.128 and 1.380 for F. verticilliodes and F. proliferatum, respectively. The linear Arrhenius-Davey model had bias factors of 0.978 and 1.002 and accuracy factors of 1.098 and 1.122 for F. verticilliodes and F. proliferatum, respectively. The results confirm the general finding that a(w) has a greater influence on fungal growth than temperature. The developed models can be applied for the prevention of Fusarium growth on maize and the development of models that incorporate other factors important to mould growth on maize.

The effect of Reuterin on Listeria monocytogenes and Escherichia coli O157:H7 in milk and cottage cheese.

A broad-spectrum reuterin produced during anaerobic fermentation of glycerol by Lactobacillus reuteri strain 12002 was found to be inhibitory and bactericidal for Listeria monocytogenes and Escherichia coli O157:H7. Lyophilized reuterin was prepared by a two-step fermentation process. A batch fermentation in a 15-liter fermentor was applied to produce a maximum biomass of L. reuteri using a modified MRS broth at pH 4.3. Further, harvested cells were used to ferment glycerol (250 mM) under anaerobic conditions. The sensitivity to reuterin of 10 strains of Listeria spp., including 6 strains of L. monocytogenes, and 6 strains of E. coli, including one enterotoxigenic E. coli strains and two enterohemorrhagic E. coli strains, was estimated. Strains of L. monocytogenes were more resistant to reuterin than E. coli strains. In cottage cheese, pH 5.4, L. monocytogenes increased by 0.4 log while E. coli O157:H7 decreased by 0.5 log in 21 days at 7 degrees C; addition of reuterin (50 to 250 units per g) to the cottage cheese reduced the viability of both organisms. The inactivation rate was more pronounced (P < or = 0.05) with E. coli O157:H7 than L. monocytogenes and it was dependent on reuterin concentration. The rate of E. coli O157:H7 population reduction reached to 2, 3, and 6 log cycles by day 7 for reuterin concentrations of 50, 100, and 150 units per g of creamed cottage cheese, respectively. While, 100, 150, and 250 units of reuterin per g caused reductions in L. monocytogenes counts by 2, 5, and greater than 5 log cycles, respectively. In UHT skim milk with 150 units of reuterin per ml, stored at 7 degrees C, the decline in the numbers of L. monocytogenes cells was higher than that in cottage cheese. Milk fat in the range of 0.5 to 3% did not affect the reuterin activity (P < or = 0.05). Addition of 3% salt enhanced the lethal effect of reuterin and diminished the initial population of L. monocytogenes by 4.5 log cycles in three days at 7 degrees C.

Effects of modified atmosphere packaging and preservatives on the shelf-life of high moisture prunes and raisins.

The growth of Aspergillus niger and Zygosaccharomyces rouxii on high moisture prunes and raisins in the presence of preservatives and packed under modified atmospheres was determined. Prunes and raisins adjusted to an aw of 0.84-0.87 in the presence of carbon dioxide atmospheres (40 and 80% CO2) did not support growth of A. niger. However, Z. rouxii spoiled the fruit samples, both in air and under CO2 conditions. Addition of low levels of K-sorbate (186 ppm in prunes and 153 ppm in raisins) or Na-benzoate (176ppm in prunes and 158ppm in raisins) delayed outgrowth of Z. rouxii. The inhibitory effect of preservatives was higher in raisins than in prunes. Modified atmospheres (40% CO2-60% N2 or 80% CO2-20% N2) combined with the addition of 417 and 343 ppm K-sorbate or 383 and 321 ppm Na-benzoate accomplished complete growth inhibition of Z. rouxii and extended the shelf-life of high moisture prunes and raisins at 30 degrees C for at least 6 months.

Prevalence of Salmonella in poultry carcasses and their products in Belgium.

During four subsequent years (1993 until 1996), a study was conducted to isolate and characterize Salmonella in poultry carcasses and their products sold in Belgium. This was a semiquantitative approach (absence per 100 cm2 or 25 cm2 or 25 g and absence per cm2 or g) to elucidate the degree of Salmonella contamination of the poultry. Serotyping was performed during the last two and a half years. Samples were frozen and kept at -20 degrees C before analysis. This may have influenced the number of Salmonella recovered. No improvement in the rate of contamination was noted during these four years, with rates being 19.4% for 1993, 24.1% for 1994, 21.9% for 1995 and 36.7% for 1996. A 100% increase of Salmonella-positive samples resulted from cutting up the carcasses into individual parts. Chicken parts were more often contaminated with Salmonella than turkey parts. Boiling hen carcasses showed the highest Salmonella contamination. Prepared poultry, chicken parts and boiling hen carcasses are sometimes associated with Salmonella contamination levels of > 1 cfu/cm2 or g. In 1996, respectively 15.1%, 4.2% and 4.2% of highly contaminated samples (> 1 cfu/cm2 or g) were found for these product groups. The predominant three serotypes were S. enteritidis (16.3%), S. hadar (15.5%) and S. virchow (14.1%). S. newport was frequently isolated from turkey products.

Growth and survival kinetics of Yersinia enterocolitica IP 383 0:9 as affected by equimolar concentrations of undissociated short-chain organic acids.

The influence of different organic acids (lactic, acetic, formic and propionic acids) at equimolar concentrations of undissociated acid with pH range of 3.9, 5.8, on the aerobic and anaerobic growth and survival kinetics of the virulent strain of Y. enterocolitica IP 383 0:9, was determined in tryptone soy broth at 4 degrees C. Growth and survival data were analyzed and fitted by a modification of the Whiting and Cygnarowicz-Provost model, using the Minpack software library. Initial generation times, initial specific growth rates, lag time and dead rate were subsequently calculated from the model parameters. The results demonstrate that the inhibitory effects of the acids were divided into two categories dependent upon pH. At high pH (5.8) the order of inhibition was formic acid > acetic acid > propionic acid > lactic acid, whereas at lower pH it became formic acid > lactic acid > acetic acid > propionic acid. The inhibitory effect of lactic acid is enhanced under anaerobic condition. Nevertheless, when the organism was cultured anaerobically, it was shown to be more tolerant to formic and acetic acids. Moreover, these variables (type of organic acid, pH and atmosphere) did not lead to the loss of the virulence plasmid in growing and surviving cells. The mechanism of inhibitory effect for each of the acids are also discussed.

Effect of water activity, modified atmosphere packaging and storage temperature on spore germination of moulds isolated from prunes.

The combined effect of medium water activity (aw) modified atmosphere packaging (MAP) on the conidial germination of Aspergillus niger, Eurotium amstelodami, Penicillium chrysogenum and Fusarium oxysporum was studied by applying response surface methodology (RSM) for 45 days incubation at 20, 30 and 40 degrees C. Oxytetracycline glucose yeast extract (OGYE) agar media were prepared over a wide range of aw (0.80-0.95), controlled by glucose. The headspace atmosphere composition varied from 0-20% for O2 and from 0-80% for CO2 with N2 as balance in the ratio medium/gas mixture of 1/3 (v/v). The results of this experiment were confirmed by repeating some combinations (aw-MAP) in the zone where the inhibition was effective. In contrast to A. niger which germinated and grew better at 30 and 40 degrees C, P. chrysogenum and F. oxysporum were capable of germinating and growing at 20 degrees C but not at 40 degrees C. E. amstelodami was less influenced by the incubation temperature than were the other species. Under anaerobic atmospheres, germination and growth of all tested species were completely inhibited at low aw, while at high aw because of residual O2, conidia germinated but failed to grow. At any aw value between 0.88 and 0.92, conidia failed to germinate when 100% CO2 was applied. Similar results were obtained for E. amstelodami and F. oxysporum in 80% CO2-20% N2 and 60% CO2-40%N2. Under aerobic conditions, at 5% O2 germination and growth occurred only at high aw, while 10 or 20% O2 combined with either 80 or 60% CO2 conidial germination and mould growth were only delayed comparing to the control (air).

Influence of modified atmosphere and preservatives on the growth of Zygosaccharomyces rouxii isolated from dried fruits.

The influence of water activity (alpha w), preservatives, modified atmosphere and their combinations on the growth of Z. rouxii was determined by cultivating two strains isolated from raisins and prunes in culture media under different conditions and by counting the colony forming units. Yeast extract glucose broths or agars were adjusted to the desired alpha w by means of glucose. Preservatives added to the media (0-600 ppm) were either K-sorbate, Na-benzoate or their mixture. Modified atmospheres were carried out by packing culture plates or flasks in plastic bags under different CO2-N2 gas mixture. Response surface design was carried out to optimize the growth inhibition of Z. rouxii by the mentioned factors. Although Z. rouxii is osmotolerant, the strains studied could not grow at alpha w 0.79. They also showed a high tolerance to CO2; even 80% CO2 seems to not inhibit growth. However, CO2 atmosphere at high pHs and low preservative concentrations stimulated yeast growth. At pH 4.0 and under modified atmosphere (80% CO2-20% N2), no growth was observed at any alpha w in the range of 0.80-0.90 when using a preservative concentration of 220 ppm Ksorbate or 280 ppm Na-benzoate.

Kinetics of interactions of lactic acid, pH and atmosphere on the growth and survival of Yersinia enterocolitica IP 383 O:9 at 4 degrees C.

The influence of different lactic acid concentrations (0.1, 0.3, 0.5, 0.7, 0.9, 1.1% v/v), within pH range of 3.9 to 5.8 on the aerobic and anaerobic growth and survival kinetics of the virulent strain of Y. enterocolitica IP 383 O:9, was determined in Tryptone Soy Broth at 4 degrees C. Growth and survival data were analyzed and fitted by a modification of the Whiting and Cygnarowicz-Provost model, using the Minpack software library. Initial generation times, initial specific growth rates, lag times and death rates were subsequently calculated from the model parameters. The stability of the virulence plasmid in growing and surviving cells was examined using crystal violet binding, low-calcium response and congo red uptake. The results demonstrate the dependancy of the growth and survival kinetics on the interaction between the three variables. The effect of lactic acid on Y. enterocolitica is greater under anaerobic than aerobic conditions. Nevertheless, the organism was found to be more tolerant of low pH conditions under anaerobic atmosphere than under an aerobic atmosphere in the absence of lactic acid. The interaction between the variables did not lead to loss of the virulence plasmid in growing or non-growing cells.

Decontamination with lactic acid/sodium lactate buffer in combination with modified atmosphere packaging effects on the shelf life of fresh poultry.

The effect of the treatment with various concentrations (2%, 5%, 7.5% and 10% w/v) of lactic acid/sodium lactate buffer (pH 3.0) combined with modified atmosphere packaging (MAP) (90% CO2/10% O2) on the shelf life and organoleptic quality of fresh chicken legs stored at 6 degrees C was investigated. The CO2 concentration of all samples packed in modified atmosphere (MA) decreased during the first 3 days of storage, followed by a gradual increase after the third day, while O2 showed a corresponding decrease. The buffering capacity of the buffer systems seem to be sufficient to maintain a low pH of the skin during storage. Legs treated with 2, 5, 7.5, and 10% (w/v) lactic acid/sodium lactate buffer (pH 3.0) combined with MAP have a shelf life at 6 degrees C of 14, 15, 16 and 17 days, respectively. The shelf life when the product was not treated with lactic acid was 1, 2.3 and 4 days shorter, respectively.

Inhibition, survival and growth of Listeria monocytogenes on poultry as influenced by buffered lactic acid treatment and modified atmosphere packaging.

The effect of the treatment with various concentrations (2%, 5% and 10% w/v) of lactic acid/sodium lactate buffer (pH 3.0), modified atmosphere (MAP) packaging (90% CO2 and 10% O2) and 10% (w/v) lactic acid/sodium lactate buffer (pH 3.0) combined with MAP on Listeria monocytogenes Z7 serotype 1 and on the shelf life of chicken legs stored at 6 degrees C was investigated. The initial contamination level of L. monocytogenes on the chicken legs surface was 8.3 x 10(2) cfu/cm2 of skin. After 2 days of storage at 6 degrees C the number of L. monocytogenes on legs treated with 2%, 5%, 10% lactic acid/sodium lactate buffer (pH 3.0) and 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP was significantly lower than the initial number of L. monocytogenes. Later, growth of L. monocytogenes was observed. After 13 days of storage at 6 degrees C the number of L. monocytogenes on legs treated with 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP was still similar to the initial number. Legs treated with 2%, 5%, 10% lactic acid/sodium lactate buffer (pH 3.0), MAP and 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP, have a shelf life at 6 degrees C of respectively 8, 9, 10, 13 and 17 days. This means a prolongation of 2, 3, 4, 7 and 11 days, respectively for storage at 6 degrees C. The antimicrobial effect of lactic acid buffer systems (pH 3.0) increased with increasing concentrations of lactic acid in the buffered system. The best results were obtained by the combined use of 10% acid/sodium lactate buffer (pH 3.0) and MAP.

The effect of treatment with buffered lactic acid on microbial decontamination and on shelf life of poultry.

The use of buffered lactic acid systems compared with unbuffered lactic acid solutions enhances the decontaminating effect and increases shelf life of chicken legs. A reduction of about 2 pH units of the chicken skin is obtained by treatment with 10% lactic acid buffer. The buffer keeps the pH of the skin lower than that of untreated legs. Legs treated with 10% lactic acid buffer have a shelf life of 12 days at 6 degrees C, which means an increase of 6 days compared with the shelf life of untreated legs.