Behaviour of Listeria monocytogenes in the presence of Listeria innocua during storage of minced beef under vacuum or in air at 0°C and 10°C

Minced beef was inoculated with low levels (1·2–1·7 log₁₀cfu g⁻¹) of Listeria monocytogenes or Listeria innocua, or a combination of the two strains. Inoculated samples were stored at 0 or 10°C under two packaging atmospheres (aerobic and vacuum) for up to 28 days and surviving organisms recovered on Palcam Agar. The only significant increases in numbers of Listeria spp. occurred in samples held at 10°C under aerobic conditions. In vacuum packs, growth of both strains was inhibited. Under aerobic conditions meat pH increased from an initial value of pH 5·85 to c. 8·85 within 28 days. The pH of vacuum packaged meat declined to c. 4·95 during the same period. These differences in pH may be related to differences in the nature and effects of different background microflora that were observed to develop under each of these packaging conditions.Pseudomonas spp. predominated in aerobically stored beef, whereas in vacuum packed beef lactic acid bacteria predominated. No significant differences were observed between the growth rates of Listeria spp. inoculated into beef mince in pure and mixed culture. This suggests that the more frequent prevalence of Listeria innocua than Listeria monocytogenes in meat and meat products is not due to overgrowth or inhibition of the pathogen (Listeria monocytogenes) by the non-pathogen(Listeria innocua) during low-temperature storage.     

Evaluation of buoyant density centrifugation as a sample preparation method for NASBA-ELGA detection ofCampylobacter jejuni in foods

Buoyant densities of four Campylobacter jejuni strains were in the range of 1·084–1·087 g ml⁻¹. Milk (3% fat) and chicken skin homogenates had buoyant densities beneath 1·033 g ml⁻¹. Discontinuous buoyant density centrifugation (BDC) in 40% Standard Isotonic BactXTractor medium respectively succeeded in recovering C. jejuni (5×10³–5×10⁴cfu ml⁻¹) from spiked milk (3% fat) and chicken skin. NASBA–ELGA detection of C. jejuni (5×10²–5×10³cfu ml⁻¹) in 12 different food samples using four different sample preparation methods was performed: RNA extraction by heating (filter and non-filter stomacher bag), RNA extraction by BDC (non-filter stomacher bag), RNA extraction by GuSCN–Triton-X-100 lysis and silica-purification (non-filter stomacher bag). BDC succeeded in overcoming inhibition of the amplification reaction except for one of the soft cheese samples. It was noticed that for chicken skin, chicken meat, pork, chicken sausage, turkey meat, milk (3% fat) and skimmed milk a simple heat treatment at 96°C without any additional precautions to prevent inhibition accomplished NASBA–ELGA detection of the pathogen. The use of a filter stomacher bag improved the quality of the NASBA–ELGA detection signal for beef but did not prevent inhibition of the amplification reaction in the cases of ground beef, prepared minced meat, soft cheese and hard cheese. The silica purification of RNA (which was used as a control treatment) accomplished NASBA–ELGA detection of C. jejuni for all of the latter food homogenates.     

Microbiological and physicochemical characteristics of Cameros cheese

Eighteen samples of Cameros cheese, a fresh Spanish goat's cheese, were collected from the four different producers of this regional kind of cheese. Physicochemical and microbiological analyses were carried out to evaluate the sanitary quality and the physicochemical characteristics of the product. The influence of the season of the year and the elaboration conditions were also studied. Cameros cheese is a fat cheese (54·2±6·5% of total solids; TS), with a high pH close to the neutrality (6·35±0·14), high moisture (TS value of 42·5±4·7%) and low salt content (0·78±0·30% of TS). Listeria monocytogenes was found in 5·6% of samples. Staphylococcus aureus counts above 2 log cfu g⁻¹were found in 55% of the cheeses studied. Thirty-three percent of the April samples and 67% of the July samples reached microbiological levels above the legally established standards. None of the samples yieldedSalmonella spp. The dairy had a significant effect on salt content (P<0·001) because different salt application methods were applied. Also significant differences (P<0·01) of S. aureus counts were detected among the different dairies. Significant seasonal differences of non-protein nitrogen (P<0·01) were detected. The season significantly affected the counts of Enterobacteriaceae (P<0·01), mesophilic micro-organisms and psycrotrophs (P<0·05) and yeast and mould counts (P<0·05).     

Effect of high pressure combined with mild heat or nisin on inoculated bacteria and mesophiles of goat's milk fresh cheese

The effect of pressures ranging from 400 to 500 MPa combined with mild heat on Staphylococcus carnosus inoculated in fresh cheese and the concurrent use of 500 MPa and nisin to inactivate cheese indigenous populations has been studied. Staphylococcus carnosus counts could not be substantially decreased with treatments at 500 MPa at 10 or 25°C for 30 min, whereas treatments at 50°C for 5 min caused a reduction of 7-log₁₀cfu g⁻¹. Multiple-cycle treatments of 500 MPa and times between 15 and 30 min also improved the inactivation rate. Combination of 500 MPa and nisin was the most effective treatment to inactivate cheese indigenous microbiota. Inactivation of Bacillus subtilis spores inoculated in fresh cheese has also been studied. Germination treatments of 60 MPa at 40°C for 210 min followed by vegetative cells inactivation treatments of 500 MPa at 40°C for 15 min caused a lethality of 4·9-log₁₀ofB. subtilis , whereas the same combination of treatments applied at 25°C only caused a 2·7-log₁₀reduction.     

Inhibition of Clostridium sporogenes growth in mascarpone cheese by co-inoculation with Streptococcus thermophilus under conditions of temperature abuse

The ability of a biological control system to inhibit the outgrowth of Clostridium sporogenes spores during storage of mascarpone cheese under temperature-abuse conditions was investigated. Challenge studies were carried out on mascarpone cheese artificially contaminated with spores of C. sporogenes (10 cfu g⁻¹), and with or without the coinoculum of a Streptococcus thermophilus strain (10⁵cfu g⁻¹). During storage at 4, 12, and 25°C, the outgrowth of clostridia spores, the growth of S. thermophilus, and the pH changes were evaluated at 10, 20, 30, and 40 days. In mascarpone cheese stored at 4° and 12°C, S. thermophilus and C. sporogenes did not show any growth. The initial pH (6·14) of the product also remained unchanged. During storage at 25°C S. thermophilus grew up to about 10⁷cfu g⁻¹after 10 days, resulting in a pH decrease of mascarpone cheese to values close to 4·5. The cell number decreased progressively during storage reaching values near to 10¹cfu g⁻¹after 40 days, whereas product acidity remained constant. C. sporogenes, when inoculated alone, also grew at 25°C. The cell number increased to levels of about 10⁷cfu g⁻¹after 20–40 days of storage according to the different mascarpone cheese lots used. No growth was found when C. sporogenes was co-inoculated in mascarpone cheese with S. thermophilus and stored at 25°C. The study on the behaviour of C. sporogenes, known as a non-toxigenic variant of Clostridium botulinum, allowed us to obtain useful information for setting up an effective biological control system to inhibit growth of the toxigenic species as well. The use of an additional barrier, besides refrigerated storage, may help to maintain the safety of mascarpone cheese in the event it was exposed to elevated temperatures.     

Decontamination of beef carcass tissue with nisin using a pilot scale model carcass washer

Nisin spray treatments were tested for controlling Gram-positive bacteria attached to beef carcass surface tissue using a pilot scale model carcass washer. Section of lean and adipose tissues were inoculated with approximately 4 log₁₀ cfu cm^-2 of Brochothrix thermosphacta, Carnobacterium divergens or Listeria innocua. Following 28°C water or nisin sprays, bacterial populations were enumerated immediately and after incubation for 24 h at 4°C. Spray treatments with water did not significantly alter the bacterial populations at day 0 or 1 (< 1 log₁₀ reduction). However, nisin spray treatments (5000 AU ml^-1) reduced populations by 1·79 to 3·54 log₁₀ cfu cm^-2 at day 0 and by 1·97 to 3·60 log₁₀ cfu cm^-2 at day 1. This study demonstrates that spray washing is an effective means of applying bacteriocins and that these compounds may be useful as sanitizers of red meat carcasses.     

The potential application of plant essential oils as natural food preservatives in soft cheese

Investigations were carried out to assess the efficiency of four plant essential oils; bay, clove, cinnamon and thyme as natural food preservatives. The effect of the plant essential oils at concentrations of 0·1, 0·5 and 1% was studied in low-fat and full-fat soft cheese against Listeria monocytogenes and Salmonella enteritidis at 4° and 10°C respectively, over a 14-day period. The composition of the cheese was shown to be an important factor in determining the effectiveness of the plant essential oils. In the low-fat cheese, all four oils at 1% reduced L. monocytogenes to ≤1·0 log₁₀cfu ml⁻¹. In contrast, in the full-fat cheese, oil of clove was the only oil to achieve this reduction. Oil of thyme proved ineffective against S. enteritidis in the full-fat cheese, yet was equally as effective as the other three oils in the low-fat cheese, reducing S. enteritidis to ≤1·0 log₁₀cfu ml⁻¹from day 4 onwards. It is concluded that selected plant essential oils can act as potent inhibitors of L. monocytogenes and S. enteritidis in a food product.     

Development of a membrane filtration method for enumeration of Listeria monocytogenes from soft cheese

A new and simple analytical method has been developed to quantify low levels (≤50 cfu g⁻¹) of Listeria monocytogenes in soft cheese. The technique allows the analysis of 1 g of cheese instead of 0·1 g or 0·01 g using the ISO 11290-2 standard method. The analysis protocol combines filtration of the decimally diluted cheese suspension through a 0·45-μm pore size cellulose ester membrane, and a culture of the filter on a Palcam agar. A tween 80–trypsin treatment used to increase filterability of cheese did not reduce L. monocytogenes counts. The tested method provided more precise results (nearer to the true value) compared to the ISO 11290-2 standard method in the enumeration of L. monocytogenes from artificially contaminated cheese. However, it improves neither repeatability nor reproducibility since the selected medium, Palcam, does not allow distinction between the different Listeria species.     

Effect of modified atmosphere and temperature abuse on the growth from spores and cereulide production of Bacillus weihenstephanensis in a cooked chilled meat sausage

The effect of modified atmosphere packaging (MAP) on the germination and growth of toxin producing psychrotolerant Bacillus spp is not well described. A model agar system mimicking a cooked meat product was used in initial experiments. Incubation at refrigeration temperature of 8 °C for 5 weeks of 26 Bacillus weihenstephanensis including two emetic toxin (cereulide) producing strains showed that B. weihenstephanensis is sensitive to MAP containing CO₂. The sensitivity to 20% CO₂ was dependent on strain and oxygen level, being increased when oxygen was excluded from the MAP. Growth from spores was observed at the earliest within 2 weeks when 20% CO₂ was combined with 2% O₂ and in 3 weeks when combined with “0”% O₂ (the remaining atmosphere was made up from N₂). Results were validated in a cooked meat sausage model for two non-emetic and one emetic B. weihenstephanensis strain. The packaging film oxygen transfer rates (OTR) were 1.3 and 40 ml/m²/24 h and the atmospheres were 2% O₂/20% CO₂ and “0”% O₂/20% CO₂. Oxygen availability had a large impact on the growth from spores in the MAP meat sausage, only the most oxygen restricted condition (OTR of 1.3 ml/m²/24 h and “0”% O₂/20 % CO₂) inhibited growth of the three strains during 4 weeks storage at 8 °C. Cereulide production was undetectable during storage at 8 °C irrespective of choice of the MAP (quantified by liquid chromatography mass spectrometry/mass spectrometry). MAP storage at 8 °C for 1 and 3 weeks followed by opening of packages and temperature abuse for 1.5 h daily at 20 °C during 1 week resulted in increased cell counts and variable cereulide production in the meat sausage. A pre-history at 8 °C for 1 week in MAP with OTR of 1.3 or 40 ml/m²/24 h and 2% O₂ resulted in cereulide concentrations of 0.816 – 1.353 µg/g meat sausage, while a pre-history under the most oxygen restricted condition (OTR of 1.3 ml/m²/24 h, “0”% O₂/20 % CO₂) resulted in minimal cereulide production (0.004 µg/g meat sausage) at abuse condition. Extension of MAP storage at 8 °C for 3 weeks followed by abuse resulted in a substantially reduced cereulide production.Data demonstrates that MAP can be used to inhibit growth of a psychrotolerant toxin producing Bacillus spp. during chill storage at 8 °C, and substantially reduce the risk of emetic food poisoning at abuse condition. Results are of relevance for improving safety of ready to eat processed chilled foods of extended durability.     

The antimicrobial effects of long-wave ultra-violet light and furocoumarins on some micro-organisms that occur in cheese brines

Listeria monocytogenes and Escherichia coli O157:H7 have been reported to survive in the brines used to store cheeses like Feta or Teleme, but such brines cannot be heat sterilized as the yeasts and lactic acid bacteria essential for normal cheese maturation are present as well. Long-wave UV light (UVA365 nm), acting in conjunction with photosensitizing compounds (e.g. furocoumarins like psoralen) might have more limited microbiocidal properties, so that, perhaps, pathogens could be eliminated from the cheese brine but not the desired yeasts and lactic acid bateria. In laboratory trials, UVA (intensity 45 W m⁻², exposure time 60 s) with psoralen (5 mg l⁻¹) was active againstListeria innocua —chosen to mimic the behaviour of L. monocytogenes—Escherichia coli O157:H7 andStaphylococcus aureus in a physiologically neutral solution, but E. coli O157:H7 (99% reduction in viable cell count) and L. innocua (99·8% reduction) were slightly less sensitive than S. aureus (99·99% reduction). Yeasts from Feta cheese brines were less affected by the same UVA/furocoumarin system—Debaryomyces hansenii (97·5% reduction) and Yarrowia lipolytica (82·7% reduction), as were typical lactic acid bacteria, namely Lactobacillus paracasei subsp. paracasei (97·8% reduction) and Lactobacillus plantarum (91·9% reduction). A UVA exposure time of 100 s with psoralen (5 mg l⁻¹) was lethal to the ‘pathogens’ but, against the desirable species, onlyYarrowia lipolytica (97·4% reduction) readily survived the same treatment. It was concluded that the UVA/furocoumarin system was microbicidal but not, at least in the form under test, sufficiently selective in its action for use with cheese brines where certain of the microfloras need to be retained.     

Nisin treatments to control Listeria monocytogenes post-processing contamination on Anthotyros, a traditional Greek whey cheese, stored at 4°C in vacuum packages

Post-processing contamination and growth of Listeria monocytogenes in whey cheeses stored under refrigeration is an important safety concern. This study evaluated commercially available nisin (Nisaplin^®) as a biopreservative to control L. monocytogenes introduced post-processing on Anthotyros, a traditional Greek whey cheese, stored at 4°C in vacuum packages for up to 45 days. The whey used (pH 6.5–6.7) was from Feta cheese manufacture, and it was subjected either to natural acidification (pH 5.3, readjusted to 6.2 with 10% NaOH) prior to heating, or to direct acidification (pH 6.0–6.2) at 80°C with 10% citric acid. Nisin was added either to the whey (100 or 500 IU g⁻¹) prior to heating, or to the cheese (500 IU g⁻¹) prior to packaging, also inoculated with ca. 10⁴ cfu g⁻¹ of L. monocytogenes strain Scott A. In cheese samples without nisin, L. monocytogenes (PALCAM agar) exceeded 7 log cfu g⁻¹ after the first 10 days of storage, irrespective of the whey acidification method. All nisin treatments had an immediate lethal effect (0.7–2.2 log reduction) on L. monocytogenes populations at inoculation (day 0), which was more pronounced with 500 IU g⁻¹ added to the whey. This treatment also suppressed L. monocytogenes growth below the inoculation level for 30 and 45 days in naturally and directly acidified samples, respectively. All other treatments had weak antilisterial effects. Nisin reversed the natural spoilage flora of Anthotyros cheese from Gram-positive to Gram-negative, and this ecological alteration was far more pronounced in the most effective antilisterial treatments.     

Destruction of acid- and non-adapted Listeria monocytogenes during drying and storage of beef jerky

Presence of Listeria monocytogenes in ready-to-eat meat products is not desired and strictly regulated in the US. Inactivation of acid- and non-adapted L. monocytogenes inoculated on beef slices was studied during drying and storage of jerky formulated with modified marinades. The inoculated (five-strain composite, c. 6·2 log cfu cm⁻²) slices were subjected to marinades (4°C, 24 h) prior to drying (60°C for 10 h) and aerobic storage (25°C for 60 days). The predrying marinade treatments tested were, first, no treatment, control (C); second, traditional marinade (TM); third, double amount of TM modified with 1·2% sodium lactate, 9% acetic acid, and 68% soy sauce containing 5% ethanol (MM); fourth, dipping into 5% acetic acid for 10 min and then applying the TM (AATM); and fifth dipping into 1% Tween 20 for 15 min and then into 5% acetic acid for 10 min followed by the TM (TWTM). Bacterial survivors on beef slices were determined during drying and storage using tryptic soy agar with 0·1% pyruvate (TSAP), and PALCAM agar. Results indicated that drying reduced bacterial populations in the order of pre-drying treatments TWTM (5·9–6·3 log cfucm⁻² in 10 h)≥AATM≥MM>TM≥C (3·8−4·6 log cfucm⁻² in 10 h). No significant (P≥0·05) difference was found in inactivation of acid-adapted and non-adapted inocula within individual treatments. Bacterial populations dropped below the detection limit (−0·4 log cfucm⁻²) as early as 4 h during drying or remained detectable even after 60 days of storage depending on acid-adaptation, predrying treatment, and agar media. These results indicated that acid-adaptation may not increase resistance to microbial hurdles involved in jerky processing and that use of modified marinades may improve the effectiveness of drying in inactivating L. monocytogenes.     

The occurrence of Listeria monocytogenes in cheese from a manufacturer associated with a case of listeriosis

A case of listeriosis was associated with the consumption of a soft cheese produced in England. Goats cheese and other products from the same food manufacturer were examined for the presence of Listeria over the following 11 months. Listeria monocytogenes was isolated from 16 of 25 cheese samples on retail sale, 12 of 24 cheese samples obtained directly from the factory, and from shelving within the plant. Phage-typing of 68 isolates of L. monocytogenes from cheese samples and the factory showed that 66 (97%) were indistinguishable from the strain isolated from the patient's cerebrospinal fluid and stool. L. monocytogenes was not isolated from seven goats milk or two yoghurt samples. Listeria innocua was isolated from 10 cheese samples, two of which contained no other species of Listeria. Levels of L. monocytogenes shortly after production were low (<10/g), but were higher (10⁵–10⁷ cfu/g) in six of the 16 cheese samples obtained from retail outlets. Multiplication of L. monocytogenes was demonstrated in cheeses contaminated at the factory and held at 4°C in the laboratory.     

Effect of lactic acid on Listeria monocytogenes and Edwardsiella tarda attached to catfish skin

This study evaluated the use of lactic acid to decontaminate Listeria monocytogenes andEdwardsiella tarda attached to catfish skin with or without mucus. At the highest inoculum levels (10⁴–10⁵cfu skin⁻¹), lactic acid (0·5–2·0%) exposure for 10 min reduced counts of L. monocytogenes firmly attached to catfish skin by 0·9–>1·9 log₁₀cfu skin⁻¹and cells loosely attached by 2·7–>3·7 logs. Counts of E. tarda firmly attached to catfish skin were reduced by 0·9–>3·0 logs and cells loosely attached by 1·5–>3·5 logs. Overall bacterial numbers of lactic acid-treated cells that were firmly attached to skin with mucus were higher than on skin without mucus. Firmly attached L. monocytogenes was more resistant to lactic acid than was firmly attached E. tarda. Catfish skin mucus decreased the antimicrobial effect of lactic acid against attached L. monocytogenes and E. tarda.     

The antimicrobial effect of thyme essential oil, nisin and their combination against Escherichia coli O157:H7 in minced beef during refrigerated storage

The antimicrobial effect of thyme essential oil (EO) at supplementation levels of 0.3%, 0.6% or 0.9%, nisin at 500 or 1000 IU/g, and their combination, on Escherichia coli O157:H7 was examined in both tryptic soy broth (TSB) and minced beef meat. 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 pathogens in minced meat. Treatment of minced beef meat with EO at 0.6% showed an inhibitory activity against E. coli O157:H7 during storage at 10 °C, but not at 4 °C. Treatment of minced beef meat or TSB with nisin at 500 or 1000 IU/g did not show any antibacterial activity against E. coli O157:H7. The combination of EO at 0.6% and nisin at 500 or 1000 IU/g showed an additive effect against the pathogen, which was higher during storage at 10 °C than at 4 °C.     

Release and identification of angiotensin-converting enzyme-inhibitory peptides as influenced by ripening temperatures and probiotic adjuncts in Cheddar cheeses

The aim of the study was to examine the release of angiotensin-converting enzyme (ACE)-inhibitory peptides in Cheddar cheeses made with starter lactococci and Bifidobacterium longum 1941, B. animalis subsp. lactis LAFTI^® B94, Lactobacillus casei 279, Lb. casei LAFTI^® L26, Lb. acidophilus 4962 or Lb. acidophilusLAFTI^® L10 during ripening at 4 and 8 °C for 24 weeks. ACE-inhibitory activity of the cheeses was maximum at 24 weeks. Cheeses made with the addition of Lb. casei 279, Lb. casei LAFTI^® L26 or Lb. acidophilus LAFTI^® L10 had significantly higher (P < 0.05) ACE-inhibitory activity than those without any probiotic adjunct after 24 weeks at 4 and 8 °C. The IC₅₀ of cheeses ripened at 4 °C was not significantly different (P > 0.05) to that ripened at 8 °C. The lowest value of the IC₅₀ (0.13 mg mL⁻¹) and therefore the highest ACE-inhibitory activity corresponded to the cheese with the addition of Lb. acidophilus LAFTI^® L10. Several ACE-inhibitory peptides were identified as κ-CN (f 96–102), α_s1-CN (f 1–9), α_s1-CN (f 1–7), α_s1-CN (f 1–6), α_s1-CN (f 24–32) and β-CN (f 193–209). Most of the ACE-inhibitory peptides accumulated at the early stage of ripening, and as proteolysis proceeded, some of the peptides were hydrolyzed into smaller peptides.     

Environmental factors affect efficacy of some essential oils and resveratrol to control growth and ochratoxin A production by Penicillium verrucosum and Aspergillus westerdijkiae on wheat grain

This study determined the efficacy of three essential oils (bay, clove and cinnamon oil) and the antioxidant resveratrol (0–500 μg g⁻¹) on the control of growth and ochratoxin A (OTA) production by Penicillium verrucosum and Aspergillus westerdijkiae (=A. ochraceus) under different water activity (a_w, 0.90, 0.95, 0.995), and temperature (15, 25 °C) conditions on irradiated wheat grain. The most effective treatment (resveratrol) was then tested on natural grain. The ED₅₀ values for growth inhibition by the oils were 200–300 μg g⁻¹ at the a_w and the temperatures tested. For resveratrol, this varied from <50 μg g⁻¹ at 0.90–0.95 a_w to >350 at 0.995a_w at both temperatures. The ED₅₀ values for the control of OTA were slightly lower than for control of growth, with approx. 200 μg g⁻¹ required for the oils and 50–100 μg g⁻¹ of the antioxidant, at 0.90/0.95a_w and both temperatures. In wet grain (0.995a_w), higher concentrations were required. For growth there were statistically significant effects of single-, two- and three-way interactions between treatments except for concentration×temperature and concentration×temperature×essential oil/antioxidant treatment. For OTA control, statistically significant treatments were a_w, temperature×a_w, concentration×temperature, treatment×concentration, and three-way interaction of concentration×a_w×treatment for P. verrucosum and A. westerdijkiae. Subsequent studies were done with the best treatment (resveratrol, 200 μg g⁻¹) on natural wheat grain with either P. verrucosum or A. westerdijkiae at 0.85–0.995a_w and 15/25 °C over 28 days storage. This showed that the populations of the mycotoxigenic species and OTA contamination could be reduced by >60% by this treatment at the end of the storage period.     

Enumeration, isolation and characterization of Bacillus cereus strains from Spanish raw rice

Bacillus cereus was present in 61 samples of raw rice analysed representing unhusked, husked and commercial origins. B. cereus in husked and white rice samples did not reach 10² cfu g⁻¹, while in the unhusked rice B. cereus densities exceeded 10³ cfu g⁻¹. Processing steps such as drying, husking and polishing reduced the number of B. cereus in the final product. Eight strains with typical morphology of B. cereus on Polymyxin–Mannitol–Egg Yolk–Phenol Red Agar (PMYPA) were isolated. According to ISO confirmatory tests, the API System tests and supplementary tests of motility, oxidase activity and enterotoxin production, these isolates were characterized and identified as B. cereus. All strains were motile, oxidase-negative and produced diarrheal enterotoxin in TSB. D and z -values were used to characterize heat resistance of spores obtained from the eight strains ofB. cereus characterized. A large diversity in heat resistance was observed among the isolates. At 90°C, D -values ranged from 2·23 to 23·26 min, with five groups of D -value means significantly different at the 95% confidence level. D₉₅- and D₁₀₀ values calculated for the eight strains ranged from 0·69 to 5·17 min and from 0·43 to 1·09 min, respectively. Statistical analysis revealed that there was significant difference between the D -value means obtained for the strains at each temperature. The z -values for the eight strains of B. cereus tested in this study ranged from 7·42°C to 8·20°C with an average of 7·7°C.     

Preliminary screening of Bifidobacteria spp. and Pediococcus acidilactici in a Swiss cheese curd slurry model system: Impact on microbial viability and flavor characteristics

A Swiss cheese curd slurry model system was used as a preliminary screening method to determine the feasibility of the incorporation of probiotic bacteria (Bifidobacterium breve R0070, Bifidobacterium infantis R0033, Bifidobacterium longum R0175, and Pediococcus acidilactici R1001) into Swiss cheese. The cheese curd was inoculated with probiotic bacteria (8.0 log₁₀ cfu g⁻¹) and ripened anaerobically for 0, 7, and 10 days at 37 °C. Following ripening, counts of the probiotic bacteria increased to 9–10 log₁₀ cfu g⁻¹, with no significant difference in the viability of the four probiotic bacteria. Viable populations of Swiss cheese background microflora in the presence of each probiotic culture were comparable with the control. Ripening time, and to a lesser extent probiotic treatment had a significant effect on the content of several volatile flavor compounds. Similarly, ripening time contributed to a significant increase in the content of a majority of the free amino acids. The study demonstrated the feasibility of the incorporation of probiotic bacteria into Swiss cheese to produce a functional food.     

Growth/survival of natural flora and Aeromonas hydrophila on refrigerated uncooked pork and turkey packaged in modified atmospheres

To evaluate the growth/survival of natural flora and Aeromonas hydrophila on refrigerated normal low (pork) and high (turkey) pH meats packaged in modified atmospheres, A. hydrophila was inoculated onto fresh pork and turkey meat slices. Inoculated and control samples were packaged in modified atmospheres (100% N₂, 20/80 and 40/60 CO₂/O₂) or in air in plastic bags and kept at 1 and 7°C. Samples packaged in air showed a similar microbiological pattern to that usually observed in fresh meat stored aerobically. Packaging in modified atmosphere produced a strong inhibition of bacterial growth at 1°C, particularly in samples stored in CO₂/O₂enriched atmospheres. Aeromonas hydrophila grew on turkey and pork meat stored in 100% N₂at 1 and 7°C. Likewise, growth of this bacterium was detected on turkey stored in 20/80 CO₂/O₂at 7°C. No growth was observed in 40/60 CO₂/O₂in any meat at both temperatures assayed.