RAT FEEDING STUDIES ON WHEAT TREATED WITH GAMMA-RADIATION. I. REPRODUCTION.
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The effect of irradiation (2 kGy) on growth of and toxin production by Staphylococcus aureus and Bacillus cereus in roast beef and gravy during storage at abuse temperatures (15 and 22 degrees C) was assessed by inoculation studies. Irradiation resulted in a 3-4 log10 reduction in numbers of both pathogens. Whenever B. cereus and S. aureus numbers reached 10(6) and 10(7) cfu/g, respectively, during storage their toxins were detectable. As the time taken to attain these levels was longer in irradiated than in unirradiated samples, toxin production by both pathogens was delayed by irradiation. When samples initially containing low levels (10(2)/g) of S. aureus were irradiated no toxin was produced during subsequent storage at 15 or 22 degrees C. Diarrhoeal toxin produced by B. cereus was detected after 2 days at 22 degrees C, but not at 15 degrees C, in samples containing 10(2) cells/g prior to irradiation. When higher numbers (10(6)/g) of either pathogen were present prior to irradiation, toxins were produced by both pathogens at 22 degrees C, but not at 15 degrees C. Microbial competition had an effect on the growth of B. cereus and S. aureus after irradiation when a low initial inoculum was applied. However, when a higher inoculum was used the pathogens outnumbered their competitors and competition effects were less important. It was concluded that low-dose irradiation would improve the microbiological safety of roast beef and gravy.
The effect of high-energy electron beam irradiation on the survival of Salmonella enterica serovar Typhimurium and psychrotrophic bacteria on commercial chicken breast meat was evaluated. Fresh chicken breast meat was purchased from a local poultry processor, inoculated with 8 log10 cfu/mL Salmonella, packaged in Styrofoam trays and over wrapped with a polyvinyl chloride film, and subjected to 0, 1, 2, or 3 kGy of irradiation. The packaged samples were stored at 4 degrees C and analyzed for Salmonella Typhimurium and psychrotrophic organisms at 0, 2, 4, 6, 8, 10, 12, and 14 d of storage. Direct plating and enrichment methods were used for S. Typhimurium analyses. The direct plating method revealed a 4 log reduction in Salmonella for chicken breasts inoculated and treated with 1, 2, or 3 kGy of irradiation. Psychrotrophic counts were conducted at 7 degrees C for 10 d and 25 degrees C for 5 d to determine the effect of incubation methods on the recovery of psychrotrophic organisms. The enrichment method resulted in the repair of injured Salmonella cells and an elevated Salmonella Typhimurium count for all irradiation dosages when compared with data reported for the direct plating method. In general, psychrotrophic counts increased as storage time increased. However, psychrotrophic counts decreased (P < 0.05) as the irradiation dosage increased.
Listeria monocytogenes did not multiply faster during storage at 7 degrees C on irradiated than on nonirradiated raw ground turkey, and there was a concentration-dependent inhibition of its multiplication by CO2. Ground turkey was gamma irradiated at 5 degrees C to 0, 1.5, and 2.5 kGy and inoculated (approximately 100 CFU/g) after irradiation with a cocktail of L. monocytogenes ATCC 7644, 15313, 49594, and 43256. The meat was then packaged in air-permeable pouches or under atmospheres containing 30 or 53% CO2, 19% O2, and 51 or 24% N2 and stored at 7 degrees C for up to 28 days. A dose of 2.5 kGy extended the time for the total plate count (TPC) to reach 10(7) CFU/g from 4 to 19 days compared to that for nonirradiated turkey in air-permeable pouches. Following a dose of 2.5 kGy at the end of the 28-day study, the TPCs were 10(6.42) and 10(4.98) under 25% and 50% CO2 atmospheres, respectively. Under air, 30% CO2, and 53% CO2 atmospheres, the populations of L. monocytogenes after 19 days incubation were 10(4.89), 10(3.60), and 10(2.67) CFU/g. The populations of lactic acid bacteria and anaerobic or facultative bacteria were also reduced by irradiation. Irradiating ground turkey did not decrease its safety when it was contaminated following processing with L. monocytogenes.
The paper handles the problem of the inactivation of the toxinogenic strain Aspergillus flavus following the application of gamma radiation to wheat. The amount of the applied dose and of the absorbed dose of ionizing radiation upon the inhibition of mycelium growth and toxin production were defined. The aflatoxin B1 was determined by extracting in chloroform and developed on Silufol R within the choroform; aceton system. The applied doses of gamma radiation (3-30 kGy) have show that the absorbed dose does not inhibit aflatoxin production. By combining the action of gamma radiation with humidity of the wheat (humidity 13-15%; 25% irradiation 6 kGy) an inactivation was reached. With the help of toxicologico-genetical tests (the Dominant Lethal Mutations Test, the Three Generations Test) the influence was traced of contaminated, irradiated substrates upon the health of experimental animals. It follows from the results obtained that in long-term feeding with contaminated wheat irradiated by gamma rays no positive mutagenic activity has been recorded. It allows to presume that wheat of humidity of 25% contaminated by a weakly toxigenic strain Aspergillus flavus irradiated by a dose of 6 kGy, and wheat of a humidity of 13-15%, contaminated by a strongly toxinogenic strain of Aspergillus flavus, irradiated by a dose of 6 kGy, are no genetic risk for white rats.
The effect of gamma-irradiation on aflatoxin B1 production by Aspergillus flavus, and the chemical composition of some different crop seeds were investigated. A. flavus infected seeds behaved differently according to their principal constituents. A. flavus caused an increase in protein and decrease in lipids and carbohydrate contents of wheat, soyabean and fababean seeds. Growth of A. flavus and production of aflatoxin B1 was inhibited at a dose level of 5 kGy. A. flavus utilizes carbohydrates of seeds for its growth and aflatoxin production. Crops were arranged, in descending order, according to aflatoxin produced in seeds as wheat > soyabean > fababean. There were no changes in chemical constituents of irradiated seeds, such as protein, lipids, and carbohydrates.
The microfloral changes on irradiated petrale sole fillets during aerobic (packaged with oxygen-permeable film), refrigerated storage were determined by the identification of bacterial and yeast isolates to the generic level. The samples were irradiated at 0.0, 0.1, 0.15, 0.2, 0.3, and 0.4 Mrad by use of a cobalt-60 gamma source, were stored at 0.5 C, and were examined periodically for spoilage, total microbial population, and composition. The preirradiation flora of the fresh fillets consisted of coryneforms, Achromobacter, Micrococcus, Flavobacterium, Pseudomonas, and Lactobacillus. Immediately after irradiation, Micrococcus, Achromobacter, coryneforms, and Bacillus were predominant. The flora of the nonirradiated fillets at the time of spoilage consisted of Pseudomonas and Achromobacter. The flora of the irradiated fillets at the time of spoilage consisted of Achromobacter and Trichosporon.
Irradiation experiments were carried out with pre-packaged sliced cooked meat products with different initial counts of Enterobacteriaceae and mesophilic aerobic bacteria. In low-sodium meat products Enterobacteriaceae could effectively be inactivated in refrigerated or frozen products by irradiation with a dose of 1 or 2 kGy respectively, provided the number of these bacteria was below 10(3) to 10(4) per g. The low-sodium meat products involved endured above treatment without being seriously affected as to sensory qualities. Shelf-life of salted cooked meat products could be prolonged by irradiating the frozen product. Irradiation with a dose of 2 kGy enabled uncooled storage of the product for a restricted period (5-7 days). Best results were obtained for meat products with a salt content in the brine phase of greater than 4.0% (w/w) and having a good hygienic quality, i.e. an initial bacterial count below 10(4)/g. Packaging in a gas atmosphere (CO2) slightly enhanced protection as compared to vacuum packaging. Off-flavours as a result of irradiation of salted cooked meat products varied from slight to strong and depended on the type of product.
Mechanically deboned chicken meat was irradiated at 0, 1.25 and 2.50 kGy (Cesium 137) and inoculated with Salmonella dublin ATCC 15480, Salmonella enteritidis ATCC 9186 or Salmonella typhimurium ATCC 14028. Samples were then stored at 5 degrees C and 10 degrees C and were subjected to microbiological analysis directly after irradiation and inoculation (time 0), and after 24, 72, 120, 168 and 216 h of storage. Samples stored at 20 degrees C were examined at time 0 and after 6, 12 and 24 h of storage. Irradiation at 1.25 and 2.50 kGy caused an average reduction in bacterial levels of 2.23 and 3.44 logs, respectively. S. dublin, S. enteritidis and S. typhimurium showed very small, insignificant changes in numbers, during storage of meat for 9 days at 5 degrees C. The final populations of S. dublin and S. typhimurium in samples irradiated before inoculation and stored at 10 degrees C or 20 degrees C were greater than the equivalent populations in samples which had not been irradiated before inoculation. Reduction of indigenous microflora in mechanically deboned chicken meat by irradiation may create better conditions for the growth of salmonellae and may thus increase the risk of salmonellosis when accidental contamination and temperature abuse occur after a radiation treatment. Therefore, irradiated mechanically deboned chicken meat should be properly refrigerated and protected against contamination.
Microbiological effects of gamma irradiation dose of 2 kGy, with and without reduction of pH to 5.3-5.2, have been investigated with a vacuum-packaged, minced meat product prepared from pork and beef with spices and cereal fillings. Either glucono-delta-lactone or ascorbic acid were used as acidulants. Experimental batches were stored at 0-2 degrees C for 4 weeks. Effect of temperature abuse condition was also studied by transferring packages for one week to 10 degrees C after 2-week holding at 0-2 degrees C. The irradiation caused two decimal reduction of the aerobic viable cell count determined after incubation at room temperature and four decimal reduction in the Enterobacteriaceae count. Lactic acid bacteria appeared to be more radiation resistant and became the dominant component of the microflora during storage. Combination of pH-reduction and irradiation prevented growth of Enterobacteriaceae even at 10 degrees C incubation.
Earlier studies conducted in our laboratory showed that heat-shocked Yersinia enterocolitica (45 degrees C for 60 min) are more resistant to a subsequent beat treatment of 55 or 60 degrees C in ground pork than cells not previously heat-shocked. The increased thermotolerance was partly attributed to the production of stress proteins. The present study was performed to determine if the stress proteins produced by heating could also afford protection to the cells to irradiation. As part of the study, the effect of air versus vacuum packaging on survival of Yersinia to irradiation was also examined. Irradiating the inoculated pork at 1.0 kGy was sufficient to completely eliminate this pathogen. The irradiation D value for both heat-shocked and non-heat-shocked cells was statistically the same (0.15 kGy). Neither heat-shocking Yersinia nor packaging under vacuum resulted in increased resistance of this organism to irradiation. In addition, no effect was seen in virulence of the cells after these treatments, when compared with controls.
Changes in aroma and coloring properties of saffron (Crocus sativus) after gamma-irradiation at doses of 2.5 and 5 kGy (necessary for microbial decontamination) were investigated. The volatile essential oil constituents responsible for aroma of the spice were isolated by steam distillation and then subsequently analyzed by gas chromatography/mass spectrometry (GC/MS). No significant qualitative changes were observed in these constituents upon irradiation, although a trained sensory panel could detect slight quality deterioration at a dose of 5 kGy. Carotene glucosides that impart color to the spice were isolated by solvent extraction and then subjected to thin-layer chromatography and high-performance liquid chromatography (HPLC). Fractionation of the above pigments into aglycon and glucosides was achieved by using ethyl acetate and n-butanol, respectively. Analysis of these fractions by HPLC revealed a decrease in glucosides and an increase in aglycon content in irradiated samples. The possibility of degradation of pigments during gamma irradiation is discussed.
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Comparisons of different models in inactivation kinetics were conducted on data obtained from high-pressure and gamma-irradiation processing. Vibrio vulnificus (MO-624) and Vibrio parahaemolyticus (O3:K6 TX-2103) suspended in phosphate-buffered saline (pH 7.4, 10(7) CFU/ml) were exposed to pressures from 207 to 379 MPa for 1 to 20 min. Inoculated whole oysters (106 CFU/g) were exposed to pressure from 276 to 379 MPa for 1 to 15 min. Pure cultures and inoculated oysters (10(6) CFU/g) also were irradiated (gamma irradiation) at doses of less than 3 kGy. Four mathematical models, the Bigelow model, Arrhenius equation, Fermi equation, and Weibull frequency distributions, were applied to microbial survival data, and performances of the different kinetic models were compared. Weibull frequency distributions can predict the high-pressure inactivation of Vibrio spp. with more accuracy in both pure cultures and inoculated oyster samples. The Fermi model provided a better description of gamma-irradiation inactivation kinetics compared with the traditional Bigelow model.
Gamma-irradiation, water activity (aw) and incubation temperature were found to affect the production of tenuazonic acid (TZA) by Alternaria alternata in tomato paste and juice. By increasing the irradiation doses, the dry weight as well as TZA decreased greatly until complete inhibition at 4 kGy. Greatest production of TZA occurred at 0.98 aw (57.5 micrograms/g and 26.3 micrograms/g) for both tomato paste and juice, respectively, at 25 degrees C. Changing temperature and aw altered the relative amounts of TZA produced in tomato paste und juice by unirradiated and irradiated conidia of A. alternata. Only trace amount of TZA was detected at 0.98 aw (1.50 micrograms/g) by 3 kGy-irradiated conidia in tomato paste, while it was inhibited completely in juice. Increasing gamma-irradiation doses and decreasing water activities decreased greatly or inhibited TZA production in both tomato paste and juice.