Growth and processing conditions affecting acid tolerance inEscherichia coli O157:H7

The impact of growth conditions (anaerobiosis, growth phase, NaCl, pH, and temperature) on the development of acid tolerance in Escherichia coli O157:H7 was investigated directly (D_pH1.15) and indirectly by monitoring the specific activity of acid phosphatase. Anaerobic growth of O157:H7 strain 43895 in synthetic rumen fluid resulted in earlier development of acid tolerance than aerobic growth. However, stationary-phase cells of both aerobic and anaerobic cultures had an equivalent degree of acid tolerance that was greater than that achieved in log-phase cultures grown anaerobically. These results are consistent with the growth-phase regulation of acid tolerance by the stationary-phase sigma factors³⁸. The addition of NaCl (1%) also enhanced acid tolerance of log-phase but not stationary-phase cells of strain 43895. Growth temperature influenced the acid tolerance with progressively greater D_pH1.15values obtained at 15, 25, and 37°1C, in both log and stationary phase. Therefore, the influence of temperature on the subsequent survival and acid tolerance of E. coli O157:H7 strain 43895 in ground beef was evaluated. Numbers of strain 43895 decreased c. 1.14 log₁₀cfu g^-1in inoculated ground beef stored at 4°1C, whereas numbers remained essentially unchanged during storage at -20°C. While pre-incubation at 15°1C for 4 h prior to storage at 4 or -20°C did not influence survival, the acid tolerance of E. coli O157:H7 survivors was significantly decreased (P<0.10001). These results indicate that the processing temperature can influence acid tolerance in E. coli O157:H7.     

Bacillus cereus emetic toxin production in cooked rice

Three mesophilic strains of Bacillus cereus known to produce emetic toxin were used to model germination, growth and emetic toxin production in boiled rice cultures at incubation temperatures ranging from 8°C to 30°C. Minimum temperatures for germination and growth in boiled rice were found to be 15°C for all strains. Toxin production at 15°C was found to be significantly greater (P<0·01; reciprocal toxin titre of 373±124) than at 20°C and 30°C (reciprocal toxin titres 112±37 and 123±41, respectively). Toxin production became detectable after 48 h incubation at 15°C, with a maximum titre reached by 96 h. At 20°C and 30°C, toxin production was detected at 24 h incubation, with a maximum titre reached by 72 h. Toxin production at 15°C was detectable at lower bacterial counts (6·2 log₁₀ cfu g⁻¹), than with incubation at 20°C and 30°C (>7·0 log₁₀ cfu g⁻¹). In this study, the lower temperature limit for germination and growth on solid laboratory medium was found to be 12°C for all strains, i.e. 3°C lower than that observed in boiled rice.     

Changes in appearance and natural microflora on iceberg lettuce treated in warm,chlorinated water and then stored at refrigeration temperature

The objective of this study was to determine the effect of warm, chlorinated water on the survival and subsequent growth of naturally occurring microorganisms and visual quality of fresh-cut iceberg lettuce. After dipping cut lettuce leaves in water containing 20 mg l⁻¹free chlorine for 90 s at 50°C, samples were stored at 5 or 15°C for up to 18 or 7 days, respectively. Populations of aerobic mesophiles, psychrotrophs, Enterobacteriaceae, lactic acid bacteria, and yeasts and molds were determined. The visual appearance and development of brown discoloration were monitored. Treatment of lettuce in warm (50°C) chlorinated water delayed browning of lettuce. Shelf life of lettuce stored at 5°C, as determined by subjective evaluation of color and general appearance, was about 5 days longer than that of lettuce stored at 15°C. Treatment in warm (50°) water, with or without 20 mg l⁻¹chlorine, and in chlorinated water at 20°C significantly (α= 0·05) reduced the initial population of mesophilic aerobic microflora by 1·73–1·96 log₁₀cfu g⁻¹. Populations increased, regardless of treatment, as storage time at 5°C and 15°C increased. The same trends were observed in populations of psychrotrophs and Enterobacteriaceae. Yeast populations increased slightly in lettuce stored at 5°C but were consistently about 3 logs lower than mesophilic aerobes. Populations of molds and lactic acid bacteria were less than 2 log₁₀cfu g⁻¹throughout storage at 5 or 15°C. Results suggest that heat (50°C) treatment may have delayed browning and reduced initial populations of some groups of micro-organisms naturally occurring on iceberg lettuce, but enhanced microbial growth during subsequent storage.     

Control of the bacteriological condition of calf brain. I. Impact of improving hygiene

Sixty calves of the Dutch Friesian (FH) breed were stunned mechanically. Without previously having been stunned, another 30 calves were stuck according to the Jewish rite. Upon opening of the skulls (1–2 h post mortem) brains of mechanically stunned calves were collected either conventionally (n = 30) or ‘hygienically’ (n = 30), i.e. using a fresh pair of surgical gloves during each removal to avoid cross contamination. For ritually slaughtered animals only the hygienic procedure was followed. Samples of 10 g were excised from undamaged hemispheres and in the mechanically stunned treatment group also from the site of impact of the captive bolt. After storage in polystyrene trays at 3 ± 1°C for 7 days sampling was repeated. Bacteriological examination included the assessment of aerobic colony counts at 30°C for 3 days (ACC-30) and 4°C for 14 days (ACC-4) and Enterobacteriaceae colony counts at 37°C for 20 h (ECC). In conventionally collected samples the ACC-30 and ACC-4 were 3.8 and 3.0 log₁₀ cfu g^?1 at day 1 and 6.2 and 6.4 log₁₀ cfu g^?1 at day 8. With hygienic collection these counts were reduced by approximately 1 log unit. Whilst by conventional practice the ECCs, at day 1 and 8 were 2.6 and 4.8 log₁₀ cfu g^?1 these counts were 1.8 and 2.6 log₁₀ cfu g^?1 for hygienic practice. In samples excised from the site of impact of the captive bolt the hygienic procedure had similar, though less marked effects. On day 1 brains from ritually slaughtered animals had a bacteriological contamination similar to that found in the hemispheres of mechanically stunned calves. However, whilst at day 8 their mean ECCs were 3.4 and 3.5 log₁₀ cfu g^?1 the percentages of plates ‘positive’ for Enterobacteriaceae were only 10% in the ritually vs. 53% in the mechanically stunned group. The Enterobacteriaceae in this case were composed of psychrotrophic non-pathogenic genera of environmental origin. Salmonella was not isolated from any sample.     

Shelf-life of Algin/Calcium Restructured Turkey Products Held under Aerobic and Anaerobic Conditions

The shelf-life of restructured products made with ground turkey or turkey breast pieces and formulated with combinations of 0.5–1.0% sodium alginate, 0.1–0.2% calcium carbonate and 0.15–0.30% lactate (ACL) was compared to product containing a combination of 1.4% NaCl and 0.32% sodium tripolyphosphate and to no-additive controls stored at 4°C under aerobic and anaerobic conditions. No significant (p > 0.05) differences were observed between ACL and salt/phosphate restructured turkey products in the rate and extent of growth of psychrotrophs or lactic acid bacteria. The salt/phosphate combination, however, repressed (p<0.05) the growth of pseudomonads. Overall, inclusion of ACL did not influence spoilage of restructured turkey meat products held under aerobic or anaerobic conditions.     

Growth of heat-treated enterotoxin-positive Clostridium perfringens and the implications for safe cooling rates

Clostridium perfringens 790-94 and 44071.C05 carrying a chromosomal and a plasmid cpe gene, respectively, were used to determine differences in heat resistance and growth characteristics between the genotypes. Heat inactivation experiments were conducted using an immersed coil apparatus. Spore germination, outgrowth, and lag phase, together named GOL time, as well as generation times were determined during constant temperatures in fluid thioglycollate (FTG) medium as well as in vacuum-packed, heat-treated minced turkey. GOL time and growth were also monitored during cooling scenarios from 65 to 10 degrees C for 3, 4, 5, 6, and 7 h in vacuum-packed, heat-treated minced turkey. Spores of strain 790-94 were approximately 10-fold more heat resistant at 85 degrees C than those of strain 44071.C05, and strain 790-94 also had a higher temperature growth range in FTG. The higher growth range for a chromosomal enterotoxin-producing CPE+ strain was confirmed using two other strains carrying a chromosomal (NCTC8239) and plasmid (945P) cpe gene. Moreover, strain 790-94 had shorter GOL times at 50 degrees C in turkey and approximately half the generation time compared with strain 44071.C05 at temperatures > or = 45 degrees C in both FTG and turkey. Strain 790-94 increased with 0.3, 1.0, 1.7, and 2.0 logs, respectively, during cooling from 65 to 10 degrees C in 4, 5, 6, and 7 h, which was significantly higher than for strain 44071.C05. A maximum acceptable cooling time of 5 h between 65 and 10 degrees C is suggested.     

Growth or Survival of Listeria monocytogenes in Ready-to-Eat Meat Products and Combination Deli Salads During Refrigerated Storage

Growth or survival of Listeria monocytogenes in cold‐smoked salmon; sliced, cooked ham; sliced, roasted turkey; shrimp salad; and coleslaw obtained at retail supermarkets stored at 5 °C, 7 °C, or 10 °C (41 °F, 45 °F, or 50 °F, respectively) for up to 14 d was evaluated. Cold‐smoked salmon, ham, and turkey were obtained in case‐ready, vacuum packages. All food products were stored aerobically to reflect additional handling within the retail supermarket. Cold‐smoked salmon, ham, and turkey supported the growth of L. monocytogenes at all 3 storage temperatures. Fitted growth curves of initial populations (about 3 log₁₀ colony‐forming units [CFU]/g) in cold‐smoked salmon, ham, and turkey stored at 5 °C achieved maximal growth rates of 0.29, 0.45, and 0.42 log₁₀ CFU/g growth per day, respectively. Storage at 10 °C increased the estimated maximal growth rate of the pathogen by 0.56 to 1.08 log₁₀ CFU/ g growth per day compared with storage at 5 °C. A decline in populations of L. monocytogenes was observed in shrimp salad and coleslaw, and the rate of decline was influenced by storage temperature. Retention of viability was higher in shrimp salad than in coleslaw, where populations fell 1.2, 1.8, and 2.5 log₁₀ CFU/g at 5 °C, 7 °C, and 10 °C, respectively, after 14 d of storage. Inability of shrimp salad and coleslaw to support the growth of L. monocytogenes may be attributed to the acidic pH (4.8 and 4.5, respectively) of the formulations used in this study. Results show that the behavior of L. monocytogenes in potentially hazardous ready‐to‐eat foods is dependent upon the composition of individual food products as well as storage temperature.     

Stability of Vacuum Cook-in-Bag Turkey Breast Rolls during Refrigerated Storage

Vacuum packaged, uncured turkey breast rolls prepared commercially in two plants by the cook-in-bag process were evaluated for physicochemical and microbiological stability during 87 days storage at 4°C. Total product weight, drained weight, pH, nonprotein nitrogen and tensile strength of turkey were not significantly affected by storage. Shear strength decreased while thiobarbituric acid numbers increased slightly in the turkey rolls. Hunter a_L color values of turkey slices varied inconsistently during storage. No psychrotrophic aerobic colony forming units were detected. Mesophilic anaerobe counts were higher in the meat core than on the surface, thus precautions should be taken to insure vacuum cook-in-bag poultry products are not temperature abused.     

Influence of four retail food service cooling methods on the behavior of Clostridium perfringens ATCC 10388 in turkey roasts following heating to an internal temperature of 74 degrees C

The influence of four food service cooling methods (CM) on growth of Clostridium perfringens ATCC 10388 in cooked turkey roasts was evaluated. Raw whole turkey roasts were inoculated with C. perfringens spores (approximately 4.23 log CFU per roast), vacuum packaged, and heated to an internal temperature of 74 degrees C. The cooked roasts were cooled as follows: whole roast cut into four quarters and held at 4 degrees C (CM1); whole roast held in a blast chiller (CM2); whole roast loosely wrapped and held at 4 degrees C (CM3); and whole roasts (three per bag) held at 4 degrees C (CM4). The roasts were analyzed for C. perfringens using Shahidi-Ferguson perfringens agar and anaerobic incubation (37 degrees C, 24 h). None of the cooling methods met the amended 2001 U.S. Food and Drug Administration Food Code guidelines for safe cooling of potentially hazardous foods. Times taken for roasts to cool from 57 to 21 degrees C using CM1, CM2, CM3, and CM4 were 2.27, 3.11, 6.22, and 8.71 h, respectively. Times taken for roasts (21 degrees C) to reach 5 degrees C ranged from 6.33 (CM1) to 19.45 h (CM4). Based on initial numbers of C. perfringens, no growth occurred in roasts cooled by CM1 or CM2, whereas numbers increased by 1.5 and 4.0 log in whole roasts cooled via CM3 and CM4, respectively. These findings indicate that certain food service cooling methods for whole cooked turkey roasts may result in proliferation of C. perfringens and increase the risk of foodborne illness by this pathogen.     

Carvacrol, cinnamaldehyde, oregano oil, and thymol inhibit Clostridium perfringens spore germination and outgrowth in ground turkey during chilling

Inhibition of Clostridium perfringens by plant-derived carvacrol, cinnamaldehyde, thymol, and oregano oil was evaluated during abusive chilling of cooked ground turkey. Test substances were mixed into thawed turkey product at concentrations of 0.1, 0.5, 1.0, or 2.0% (wt/wt) along with a heat-activated three-strain C. perfringens spore cocktail to obtain final spore concentrations of ca. 2.2 to 2.8 log CFU spores per g of turkey meat. Aliquots (5 g) of the ground turkey mixtures were vacuum packaged and then cooked in a water bath, where the temperature was raised to 60 degrees C in I h. The products were cooled from 54.4 to 7.2 degrees C in 12, 15, 18, or 21 h, resulting in 2.9-, 5.5-, 4.9-, and 4.2-log CFU/g increases, respectively, in C. perfringens populations in samples without antimicrobials. Incorporation of test compounds (0.1 to 0.5%) into the turkey completely inhibited C. perfringens spore germination and outgrowth (P < or = 0.05) during exponential cooling in 12 h. Longer chilling times (15, 18, and 21 h) required greater concentrations (0.5 to 2.0%) to inhibit spore germination and outgrowth. Cinnamaldehyde was significantly (P < 0.05) more effective (<1.0-log CFU/g growth) than the other compounds at a lower concentration (0.5%) at the most abusive chilling rate evaluated (21 h). These findings establish the value of the plant-derived antimicrobials for inhibiting C. perfringens in commercial ground turkey products.     

Behavior of Shiga-toxin-producing Escherichia coli in ewe milk stored at different temperatures and during the manufacture and ripening of a raw milk sheep cheese (Zamorano style)

This study was conducted to assess the survival of 2 wild Shiga toxin-producing Escherichia coli strains (one serotype O157:H7 and one non-O157:H7) in ewe milk stored at different conditions and to examine the fate of the O157 strain during the manufacture and ripening of a Spanish sheep hard variety of raw milk cheese (Zamorano). The strains were selected among a population of 50 isolates, which we obtained from ewe milk, because of their high resistance to 0.3% lactic acid. Both strains were inoculated (approximately 2 log₁₀ cfu/mL) in raw and heat-treated (low-temperature holding, LTH; 63°C/30 min) ewe milk and stored for 5 d at 6, 8, and 10°C and also according to a simulation approach for assessing the effects of failures in the cold chain. The minimum growth temperature for the O157:H7 strain in LTH and raw ewe milk was 8°C. For the non-O157:H7 strain, the lowest temperature showing bacterial growth in LTH ewe milk was 6°C, but it did not grow at any of the tested conditions in raw milk. It appears that the O157 strain was more susceptible to cold stress but was likely a better competitor than the non-O157 strain against the milk autochthonous microbiota. For manufacture of Zamorano cheese, raw milk was inoculated with approximately 3 log₁₀ cfu/mL, and after 2 mo of ripening at 10 to 12°C, the cheeses showed the expected general characteristics for this variety. The O157:H7 strain increased 0.9 log₁₀ cfu/g after whey drainage and during ripening and storage decreased by 2.9 log₁₀ cfu/g. Nevertheless, its detectable level (estimated at 6.2 cfu/g) after 2 mo of ripening suggests that Zamorano cheese manufactured from raw ewe milk contaminated with E. coli O157:H7 could represent a public health concern.     

Survival of Campylobacter coli in porcine liver

Enteropathogenic Campylobacter jejuni, Campylobacter coli and Campylobacter lari are currently the most common causes of acute infectious diarrhoeal illness in the UK and in most developed countries. Many domestic animals, including pigs, act as natural reservoirs of these organisms and infection may occur through the ingestion of contaminated foodstuffs. Therefore, the safety of porcine liver produced in Northern Ireland was assessed in relation toCampylobacter spp. Storage trials showed that Campylobacter spp. were not able to proliferate in liver at 37°C, but could persist at 4°C and 15°C. Survival was better, however, during storage at 4°C than at 15°C.Campylobacter were rapidly killed in raw liver homogenates and distilled water at 37°C, but not at 4°C. An initial inoculum of 8 log₁₀cfu g⁻¹C. coli was undetectable in liver homogenates after 24h storage at 37°C. Campylobacter coli were sensitive to freezing on liver slices at −18°C and were reduced by 5 log₁₀cycles after 7 days storage. Cells survived better on chilled liver slices and in autoclaved liver homogenates than in raw liver homogenates at all temperatures, which indicates the presence of a heat-labile antagonistic agent in raw liver homogenates. Growth and survival of C. coli was not affected by Lactobacillus plantarum, as C. coli was able to reach 8·5 log₁₀cfu ml⁻¹in 7 days and maintain its viability in the presence of 8·0 log₁₀cfu ml⁻¹L. plantarum. Thus, storage of C. coli on porcine liver at 4°C selected for the survival of this pathogen compared to similar storage at 37°C.Such information may be useful in identifying conditions and treatments that could be integrated in HACCP strategies, or be used to design processes that prevent proliferation and/or destroy Campylobacter spp. that may be present in liver.     

Development of a spray-dried conjugated whey protein hydrolysate powder with entrapped probiotics

Bifidobacterium animalis ssp. lactis ATCC27536 and Lactobacillus acidophilus ATCC4356 were encapsulated in a conjugated whey protein hydrolysate (WPH10) through spray drying. Probiotic cultures were added at the ratio of 1:1 into the conjugated WPH10 solution at a spiking level of about 10 log₁₀ cfu/mL. The mixture was spray dried in a Niro drier with inlet and outlet temperatures of 200°C and 90°C, respectively. The final dried product was determined for cell viability and further stored for 16 wk at 25°, 4°, and ?18°C to monitor viability and functionality. Micro images showed the presence of link bridges in non-conjugated WPH10, whereas, in the case of conjugated WPH10, round particles with pores were observed. The mean probiotic counts before and after spray drying were 10.59 log₁₀ cfu/mL and 8.98 log₁₀ cfu/g, respectively, indicating good retention of viability after spray drying. The solubility and wetting time of the WPH10-maltodextrin (MD) encapsulated probiotic powder were 91.03% and 47 min, whereas for WPH10, the solubility and wetting time were 82.03% and 53 min, respectively. At the end of storage period, the counts were 7.18 log₁₀ cfu/g at 4°C and 7.87 log₁₀ cfu/g at ?18°C, whereas at 25°C the counts were significantly reduced, to 3.97 log₁₀ cfu/g. The solubility of WPH-MD powder was 82.36%, 83.1%, and 81.19% at ?18°C, 4°C, and 25°C, respectively, and wetting times were 61 min, 60 min, and 63 min at ?18°C, 4°C, and 25°C, respectively. By contrast, for WPH10 powder, the solubility significantly reduced to 69.41%, 69.97%, and 68.99% at ?18°C, 4°C, and 25°C, and wetting times increased to 71 min, 70 min, and 72 min at ?18°C, 4°C, and 25°C, respectively. The conjugated WPH10 is thus demonstrated as a promising carrier for probiotics and can be further used as an ingredient for developing functional foods, to harness their enhanced functionality and health benefits derived from both WPH and probiotics.     

INFLUENCE OF MODIFIED ATMOSPHERE PACKAGING ON GROWTH OF CLOSTRIDIUM PERFRINGENS IN COOKED TURKEY

Clostridium perfringens containing samples of sterile ground turkey were studied to assess growth under modified atmosphere conditions. Samples were packaged under various atmospheres (CO₂/O₂/N₂: 75/5/20, 75/10/15, 75/20/5, 25/20/55, 50/20/30), stored at 4, 15 and 28C, and sampled periodically for growth. Diluted samples were plated on Shahidi Ferguson perfringens agar (Difco Laboratories, Detroit, MI) to determine vegetative cell counts. Temperature abuse (cyclic and static) of the turkey product was also investigated. The results showed that the growth of C. perfringens was slowest under 25–50% CO₂/20% O₂/balance N₂ at 15 and 28C. There was no growth at 4C for up to 28 days. Temperature abuse (28C storage) of refrigerated products for 8 h did not permit C. perfringens growth. Use of 25–50% CO₂/20% O₂/balance N₂ may extend the shelf-life of turkey, but in the absence of proper refrigeration, it cannot be relied upon to eliminate the risk of C. perfringens food poisoning .     

Quality of minimally processed carrots as affected by warm water washing and chlorination

Different applications of cold and warm tap water (4 °C and 50 °C) with and without chlorination, respectively, in the washing of uncut peeled carrots (Daucus carota L.) were conducted, and their effects on sensory and microbiological properties during storage for 9 days at 4 °C were assessed. To minimise cross-contamination of almost sterile inner root parts with the highly contaminated outer cortex during processing the peeled carrots were washed prior to cutting. The washing treatments were carried out using a commercial processing line, thus facilitating the scale-up to industrial production. Populations of aerobic mesophilic bacteria, lactic acid bacteria and enterobacteria on these minimally processed carrots were determined, and the sensorial quality of shredded carrots was evaluated by a sensory panel throughout storage. Additionally, colour, texture, sugars, and trichloromethanes were analysed. Washing uncut carrots with cold chlorinated water (200 mg/l, 4 °C) and warm tap water (50 °C) ensured sugar retention and reduced aerobic mesophilic bacteria by 1.7 and 2.0 log₁₀ colony forming units per gram (cfu/g), respectively, while washing with warm chlorinated water (200 mg/l) resulted in a 2.3 log₁₀ cfu/g reduction. By-product formation due to chlorination was negligible. Sensorial properties of the latter samples were slightly affected. It was shown that both washing uncut knife-peeled carrots with cold chlorinated water (4 °C) and warm tap water (50 °C), respectively, provided good microbiological safety paired with improved sensorial properties. Moreover, fresh-like character of the products was retained, as indicated by the persisting respiration of the living tissues.Industrial relevanceMinimally processed vegetables are an increasing market. Shelf-life extension and consumer safety are of immense relevance for the fresh-cut industry; therefore, the application of antimicrobial agents such as chlorine is widespread. However, various consumer groups object to the use of chlorinated water. The present study aimed at comparing the efficacy of cold and warm tap water with and without chlorination, respectively, in washing uncut carrots during the production of shredded, packaged carrots while operating on pilot-plant scale under conditions of industrial practice. In view of microbial reduction and maintenance of sensory properties, the use of cold chlorinated water and warm tap water, respectively, proved to be effective for washing peeled carrots. By-product formation due to chlorination was negligible.     

Delayed Clostridium perfringens growth from a spore inocula by sodium lactate in sous-vide chicken products

Clostridium perfringens growth from a spore inoculum was investigated in vacuum-packaged, cook-in-bag marinated chicken breast that included 0%, 1.5%, 3%, or 4.8% sodium lactate (NaL; w/w). The packages were processed to an internal temperature of 71.1 degrees C, ice chilled and stored at 4, 19, and 25 degrees C. The total C. perfringens population was determined by plating diluted samples on Tryptose-sulfite-cycloserine agar followed by anaerobic incubation for 48 h at 37 degrees C. At 25 degrees C, addition of 1.5% NaL was effective in delaying growth for 29 h. Increasing the NaL level to 4.8%, C. perfringens growth from a spore inoculum during storage at 25 degrees C for 480 h was not observed. At 19 degrees C, the growth was > 6 log 10 cfu/g by 288 h in control samples. In samples with 3.0% or 4.8% NaL, the growth of C. perfringens from spores was dramatically restricted with little or no growth in 648 h at 19 degrees C. C. perfringens growth was not observed at 4 degrees C regardless of NaL concentration. The D-values at 55 degrees C ranged from 47.40 (no NaL) to 57.58 min (1.5% NaL). Cyclic and static temperature abuse of refrigerated products for 20 h did not permit C. perfringens growth. However, temperature abuse of products for periods 24 h or longer in the absence of NaL led to growth of C. perfringens from a spore inoculum. An extra degree of safety may be assured in such products by supplementation with NaL at 1.5-4.8% NaL level.     

Anaerobic Microbiology of Fresh Beef Packaged Under Modified Atmosphere or Vacuum

Beef was packaged under CO₂ containing < 500 ppm O₂ (MAP), vacuum, or air and stored at 0, 2, or 4°C. Samples were analyzed weekly for bacterial numbers using anaerobic plate count (pre-reduced medium and anaerobic chamber), -aerobic plate count, and anaerobic jar plate count (MAP samples only) methods. For both MAP and vacuum packaged samples, the anaerobic plate count was consistently greater than the aerobic plate count and for MAP samples the anaerobic plate count was consistently higher than the anaerobic jar plate count. Differences between plating methods were most frequent during the latter third of 0 and 2°C storage. Anaerobic isolates from MAP samples were most often lactic acid cocci and staphylococci. No clostridia were isolated from any of the treatments.     

Spore-forming bacteria in commercial cooked,pasteurised and chilled vegetable purées

In commercial purées of broccoli, carrot, courgette, leek, potato and split pea, pasteurized in their final packaging and analysed at two periods, Bacillus spp. were the dominant aerobic mesophilic bacteria (AMB). Initial numbers were generally lower than 2 log cfu g⁻¹. They increased up to 6–8 log cfu g⁻¹after about 20 days of storage at 10°C. At 4°C, numbers of AMB after 20 days were lower than 3 log cfu g⁻¹in potato purée, lower than 4 log cfu g⁻¹in leek purée, and between 3 and 6 log cfu g⁻¹in other products. Strict anaerobes were in markedly lower numbers than AMB. At all storage temperatures tested courgette purée usually showed the most rapid bacterial growth and spoilage. On this product, an increase in storage temperature from 4°C to 10°C resulted in a threefold reduction in time to 5 log cfu g⁻¹, and time to spoilage. Growth kinetics of AMB in courgette purée at 20°C, 15°C, 10°C, 6·5°C and 4°C were determined using a mathematical model. Three hundred and forty eight isolates were identified using the API system. Bacillus circulans, B. macerans and B. polymyxa were among the main species isolated from products stored at 4°C and 10°C, while B. subtilis and B. licheniformis were the dominant species in product stored at abuse temperature. Bacillus cereus was isolated from all storage conditions, but mostly from products stored at abuse temperature.     

Increase in aflatoxins due to Aspergillus section Flavi multiplication during the aerobic deterioration of corn silage treated with different bacteria inocula

The growth of Aspergillus flavus and the production of aflatoxins (AF) during the aerobic deterioration of corn silage represent a problem for animal and human health. This experiment was conducted to evaluate whether the presence of A. flavus and AF production originate from the field or additional AF are produced during the fermentation phase or during aerobic deterioration of corn silage. The trial was carried out in northern Italy on corn at a dry matter (DM) level of 34%. The fresh herbage was either not treated (C) or treated with a Lactobacillus buchneri (LB) NCIMB 40788 [(at 3 × 10⁵ cfu/g of fresh matter (FM)], Lactobacillus hilgardii (LH) CNCM I-4785 (at 3 × 10⁵ cfu/g of FM), or their combination (LB+LH; at 1.5 × 10⁵ cfu/g of FM of each strain) ensiled in 20-L silos and opened after 250 d of ensiling. After silo opening, the aerobic stability was evaluated and samples were taken after 7 and 14 d of air exposure. The pre-ensiled material, the silages at silo opening, and the aerobically exposed silages were analyzed for DM content, fermentative profiles, microbial count, nutritive characteristics, DM losses, and AFB₁, AFB₂, AFG₁, and AFG₂ contents. Furthermore, a subsample of colonies with macromorphological features of Aspergillus section Flavi was selected for AF gene pattern characterization and in vitro AF production. The presence of A. flavus was below the detection limit (<1.00 log₁₀ cfu/g) in the fresh forage before ensiling, whereas it was found in 1 out of 16 silage samples at silo opening at a level of 1.24 log₁₀ cfu/g. The AF were found in both the fresh forage and at opening in all the samples, with a predominance of AFB₂ (mean value of 1.71 μg/kg of DM). The inoculation of lactic acid bacteria determined a reduction in the lactic-to-acetic ratio compared with the control. A larger amount of acetic acid resulted in a lower yeast count and higher aerobic stability in the treated silages than in the control ones. At the beginning of aerobic deterioration, the yeasts increased to over 5 log₁₀ cfu/g, whereas the molds were close to the value observed at silo opening. When the inhibiting conditions were depleted (pH and temperature higher than 5 and 35°C, respectively), both the total molds and A. flavus reached higher values than 8.00 and 4.00 log₁₀ cfu/g, respectively, thus determining the ex novo production of AFB₁ during aerobic deterioration, regardless of treatments. The analysis of gene pattern showed that 64% of the selected colonies of A. flavus showed the presence of all 4 AF gene patterns, and 43% of the selected colonies were able to produce AF in vitro. During air exposure, after 1,000°C·h have been cumulated, starch content decreased (below 10% DM) and concentration of neutral detergent fiber, acid detergent fiber, hemicelluloses, crude protein, and ash increased. The inoculation with LB and LB+LH increased the aerobic stability of the silages and delayed the onset of aerobic microbial degradation, which in turn indirectly reduced the risk of A. flavus outgrowth and AFB₁ production after silage opening.     

Listeria monocytogenes Inactivation in Turkey Rolls and Battered Chicken Nuggets Subjected to Simulated Commercial Cooking

Cured and uncured turkey rolls inoculted with 10⁷Listeria monocytogenes CFU/g were vacuum packaged and cooked to internal temperatures of 68°C and 74°C, respectively, in a steam-injected chamber. Samples were stored up to 15 wk at 4°C. Battered chicken nuggets were also inoculated internally with about 10⁷L. monocytogenes CFU/ g. Nuggets enclosed in bags were cooked under moist heating conditions in a convection oven to an internal temperature of 71°C. Nuggets were flushed with 30% CO₂, 70% N₂ atmosphere and sealed. Chicken nuggets were stored at 4°C up to 30 days. No Listeria monocytogenes were recovered from the cooked products suggesting that similar commercial processes are adequate to reduce populations of L. monocytogenes below detection limits.