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Ionizing radiation sensitivity of Listeria monocytogenes ATCC 49594 and Listeria innocua ATCC 51742 inoculated on endive (Cichorium endiva).

Ionizing radiation inactivates the pathogenic bacteria that can contaminate leafy green vegetables. Leaf pieces and leaf homogenate of endive (Cichorium endiva) were inoculated with the pathogen Listeria monocytogenes (ATCC 49594) or Listeria innocua (ATCC 51742), a nonpathogenic surrogate bacterium. The radiation sensitivity of the two strains was similar, although L. innocua was more sensitive to the type of suspending leaf preparation. During refrigerated storage after irradiation, the population of L. monocytogenes on inoculated endive was briefly suppressed by 0.42 kilogray (kGy), a dose calibrated to achieve a 99% reduction. However, the pathogen regrew after 5 days until it exceeded the bacterial levels on the control after 19 days in storage. Treatment with 0.84 kGy, equivalent to a 99.99% reduction, suppressed L. monocytogenes throughout refrigerated storage. Doses up to 1.0 kGy had no significant effect on the color of endive leaf material, regardless of whether taken from the leaf edge or the leaf midrib. The texture of leaf edge material was unaffected by doses up to 1.0 kGy, whereas the maximum dose tolerated by leaf midrib material was 0.8 kGy. These results show that endive leaves may be treated with doses sufficient to achieve at least a 99.99% reduction of L. monocytogenes with little or no impact on the product's texture or color.

Asteraceae↗

Combined effects of ionizing-irradiation and different environments on Clostridium botulinum type E spores.

We examined the combined effects of gamma-radiation (24 degrees C) on spores of Clostridium botulinum-type Eklund strain suspended in different gas-saturated Na-phosphate buffer in absence or presence of protectors or sensitizers. Response surface methodology (RSM) was also used to ascertain the effects of radiation on the recovery of spores using a medium containing various levels of NaCl or Na-thioglycollate. The former (< 0.5%) decreased viable spore counts, but the latter (0.15%) did not. Irradiation inactivation of Eklund spores was most effective in air-saturated buffers compared to N2O and N2 gas. The Na2-EDTA (0.01 M) was the most efficient radioprotector of spores due to its reactivity toward hydroxy radicals, followed by t-butanol (0.1 M) in NO2 or N(2)-saturated buffers, respectively. Catalase (10.0 mg ml(-1)) and DL-cysteine (0.1 mM) sensitized the spores during irradiated N2O or N(2)-saturated buffers, and NaCl (0.01 M) only sensitized spores in N2 environment. Spores frozen at -75 degrees C for 30 days and thawed prior to use were more sensitive to radiation damage compared to freshly prepared spores. Glycerol (15%), in Na-phosphate buffer (pH 7.0, 0.06 M), protected Eklund spores and increased the number of spores from 10(6) to 10(11) colony forming unit (CFU) ml(-1), and enhanced their radiosensitivities. Seven strains of C. botulinum type E were screened for plasmids and strain BL764 had two plasmids (15.8 and 46.8 mDa), BL4028 also had two (4.4 and 13.2 mDa), BL4850 contained only one (4.9 mDa), whereas EQA, BL211, Eklund, and Beluga had none. Gamma-Radiation (10 kGy, absorbed dose) cured the 15.8-mDa plasmid in strain BL764, but its absence yielded no changes in toxigenicity.

Clostridium botulinum↗

Consumer acceptance of irradiated poultry.

A simulated supermarket setting (SSS) test was conducted to determine whether consumers (n = 126) would purchase irradiated poultry products, and the effects of marketing strategies on consumer purchase of irradiated poultry products. Consumer preference for irradiated poultry was likewise determined using a home-use test. A slide program was the most effective educational strategy in changing consumers' purchase behavior. The number of participants who purchased irradiated boneless, skinless breasts and irradiated thighs after the educational program increased significantly from 59.5 and 61.9% to 83.3 and 85.7% for the breasts and thighs, respectively. Using a label or poster did not increase the number of participants who bought irradiated poultry products. About 84% of the participants consider it either "somewhat necessary" or "very necessary" to irradiate raw chicken and would like all chicken that was served in restaurants or fast food places to be irradiated. Fifty-eight percent of the participants would always buy irradiated chicken if available, and an additional 27% would buy it sometimes. About 44% of the participants were willing to pay the same price for irradiated chicken as for nonirradiated. About 42% of participants were willing to pay 5% or more than what they were currently paying for nonirradiated chicken. Seventy-three percent or more of consumers who participated in the home-use test (n = 74) gave the color, appearance, and aroma of the raw poultry products a minimum rating of 7 (= like moderately). After consumers participated in a home-use test, 84 and 88% selected irradiated thighs and breasts, respectively, over nonirradiated in a second SSS test.

Animals↗

Juice irradiation with Taylor-Couette flow: UV inactivation of Escherichia coli.

A novel reactor is described with flow characteristics that approach that of ideal plug flow but with a residence time that is uncoupled from the hydrodynamics or boundary layer characteristics. The design described consists of an inner cylinder that rotates within a stationary but larger outer cylinder. At low rotation rates, a laminar, hydrodynamic configuration called Taylor-Couette flow is established, which consists of a system of circumferential vortices within the annular fluid gap. The latter constitutes a spatially periodic flow that is the hydrodynamic equivalent to cross flow over a tube bank or lamp array. These vortices provide radial mixing, reduce the boundary layer thickness, and are independent of the axial flow rate and thus the fluid residence time. An additional feature of the rotating design is the repetitive exposure of the fluid parcels to a minimum number of lamps, which substantially reduces the maintenance requirements. Inactivation data for Escherichia coli (ATCC 15597) were recorded in commercial apple and grape juice that are relatively opaque to UV radiation. With initial E. coli concentrations of approximately 10(6) CFU/ml, Taylor-Couette flow was found to provide a 3- to 5-log improvement in the inactivation efficiency compared with simple channel flow between concentric cylinders.

Beverages↗

Irradiation of chewable tobacco mixes for improvement in microbiological quality.

Microbiological quality of chewable tobacco mixes traditionally known as "Gutkha" was studied. The microbiological analysis of 15 samples analyzed revealed high bacterial and fungal counts. The total viable counts were in the range of 1.8 x 10(4) to 7.2 x 10(4) CFU g(-1) and the yeast and mold count from 3.6 x 10(3) to 7.1 x 10(4) CFU g(-1). The proteolytic and lipolytic counts were 9 x 10(2) to 2.6 x 10(3) CFU g(-1) and 2.6 x 10(3) CFU g(-1), on an average, respectively. Lecithinase-positive Staphylococcus aureus was found in 2 of the 15 samples analyzed; the counts were up to 3.4 x 10(3) CFU g(-1). Coliform and Salmonella spp. were found to be absent. Aflatoxins B , B2, and G2 were found to be present in all the samples. These samples were exposed to gamma radiation (60Co) at 1-, 2-, 3-, 5-, 10-, and 25-kGy doses. The decrease in total viable count and fungal count was noticed with increase of radiation dose. The 3-kGy dose was observed to be the sterilization dose for Gutkha. At this dose no survival of organisms was noticed and no revival was observed during postirradiation storage at room temperature for 6 months.

Aflatoxins↗

Photostimulated luminescence detection of irradiated herbs, spices, and seasonings: international interlaboratory trial.

An interlaboratory trial was conducted to validate photostimulated luminescence (PSL) methods for herbs, spices, and seasonings. Forty products (11 herbs, 17 spices, and 12 seasonings) were purchased from a local commercial source, and randomly selected samples were irradiated with 10 kGy. Four blended products were prepared at Scottish Universities Research and Reactor Centre, mixing varying proportions of irradiated material with the untreated product. Precharacterization against a predefined threshold identified low sensitivity products (black and white peppers) and products with high natural signals (thyme, sage, parsley, and mixed herbs), both of which might be susceptible to misclassification. Precharacterization also revealed whether calibration was likely to resolve overlap between classification categories. Eight sets of screening data and 5 sets of calibrated data were returned by participants. Of the 840 samples sent, 1593 screening measurements and 788 calibrated measurements were received from 662 samples. In screening mode, participants reached definitive conclusions in 87% of cases, 99% of which were correct. Of the remaining 13%, calibration to identify low-sensitivity resolved 60% of cases. Overall, 94% of samples were correctly identified by either screening alone, or screening plus calibration; 6% remained unclassified and therefore required further investigation by thermoluminescence. The results confirm the validity of the PSL method for herbs, spices, seasonings, and blends, and emphasize the need for calibration to identify low-sensitivity samples. This method has now been adopted by the European Committee for Standardization (CEN) and the Codex Alimentarius Commission.

Calibration↗

The growth of Salmonella typhimurium on irradiated, raw, skinless chicken breast.

Salmonella typhimurium was inoculated onto 1 g cubed samples of irradiated, raw, skinless chicken breast, which were then incubated at 30 degrees C under humid conditions. Kinetic growth data was obtained by means of viable counts performed on triplicate samples over a 24 h period. In addition, the spatial arrangement of cells on samples taken 6, 12 and 24 h after inoculation was observed by scanning electron microscopy. The population entered exponential growth approximately 3 h after inoculation, and maintained a constant rate of growth for approximately 13 h before entering a stationary phase. A generation time of 0.74 h was recorded. Scanning electron microscopy observations revealed colonial development from loose clusters of cells at 6 h to discrete, compact microcolonies (approximately 40 microns in diameter) by 12 h. By 24 h colonies were well-developed (approximately 600-700 microns in diameter) with a well-defined colony periphery. The results of this study give insight into the growth and development of bacteria on meat tissue, and serve to highlight that the nature of such growth is quite different from that in dispersed liquid culture systems, i.e. those traditionally used to model such growth.

Animals↗

Productive performance of broiler chicks fed diets containing irradiated meat-bone meal.

Experiments were carried out to study the changes in the values of feed efficiency (FE), total protein efficiency (TPE) and metabolizable energy efficiency (MEE) of broiler chicks fed during four age periods (14-21, 21-28, 28-35 and 35-42 days). Five experimental diets were similar in composition but different in the irradiation dose (0, 5, 10, 25 or 50 kGy) applied to the meat-bone meal content of each diet. The results indicated that feeding of broiler chicks on irradiated meat-bone meal (5-50 kGy) had no significant (P>0.05) effects on FE, TPE, MEE and body weight gain during the four experimental age periods. The average values of the FE (feed consumption/weight gain), TPE (weight gain/total protein consumption) and MEE (metabolizable energy consumption in MJ/kg gain) were 1.96, 2.59 and 23.13, respectively. The values of FE and MEE increased while TPE decreased significantly (P<0.05) with age. The pooled FE value was 1.60 during the 14-21 days of age and increased to 2.66 from 35-42 days of age. However, the TPE value was 3.01 during the 14-21-day age period and decreased to 1.82 over the 35-42-day age period. The energy consumption value to produce one kg of body weight increased during the 35-42-day period by 7.71-12.72 MJ/kg gain in comparison with that from the other experimental age periods. Weight gain rate decreased significantly (P<0.05) during the 35-42-day period (66 g/week/bird) when compared with the rate observed during the 14 through 35-day age period.

Age Factors↗

Absorption of methionine and 2-hydroxy-4-methylthiobutoanic acid in conventional and germ-free chickens.

The apparent absorption of 3H-labeled L-Met and L-2-hydroxy-4-methylthiobutoanic acid (MHA-FA) was compared in germ-free and conventional broiler chickens to determine the effect of intestinal bacteria on the absorption of Met and MHA-FA. The two diets contained 0.236% of added Met or MHA-FA. Nineteen germ-free birds were maintained in two isolators and fed diets that had been sterilized by gamma irradiation (50 kilogreys). Nineteen conventional birds were reared in batteries and received nonirradiated feed. Diets were fed ad libitum for 3 wk. On d 21 of the experiment, the birds fasted overnight and were refed the experimental diets to which 1.11 x 10(7) Bq of 1-[methyl3H]MHA-FA or 1-[methyl3H]Met/kg of feed had been added. 51CrCl3 (1.11 x 10(7) Bq/kg of feed) was added as an indigestible marker. After 3 h the birds were euthanized, and their intestinal tracts were removed and partitioned into six sections. Residual Met and MHA-FA in digesta were calculated as the ratio of 3H:51Cr in each sample divided by the ratio of 3H:51Cr in the feed. The residual MHA-FA in the distal ileum of germ-free broilers was lower than in conventional birds (4.7 and 10.2% respectively; P < 0.05). In contrast the residual Met in the distal ileum of germ-free broilers was not different than in conventional birds (3.0 and 3.7% respectively; P > 0.05). This study demonstrates that intestinal bacteria significantly reduce the apparent absorption of MHA-FA from the intestinal tract of broiler chickens.

Animal Feed↗

Comparison of three methods of DNA extraction from cold-smoked salmon and impact of physical treatments.

AIMS: To compare three bacterial DNA extraction procedures on cold-smoked salmon (CSS) and assess the impact on their efficiency of two physical treatments of the food matrix, ionizing irradiation and freezing. METHODS AND RESULTS: As molecular methods for bacterial detection have become an important analytical tool, we compared bacterial DNA extraction procedures on CSS. Working with frozen and irradiated CSS, we obtained negative responses from samples known to be highly contaminated. Thus, we decided to study the impact of these two physical treatments on bacterial DNA extraction procedures. The efficiency of bacterial DNA extraction directly from the fish matrix suspension was measured by an rpoB PCR-based reaction. The results demonstrated that the DNeasy tissue extraction kit (Qiagen, Courtaboeuf, France) was the most efficient and reproducible method. We also showed that freezing and ionizing irradiation have a negative impact on DNA extraction. This was found probably not to be due to inhibition as the PCR reaction remained negative after adding BSA to the PCR mix reaction. CONCLUSIONS: The extraction kit was the most efficient method. Physical treatments were shown to hamper bacterial DNA extraction. SIGNIFICANCE AND IMPACT OF THE STUDY: Attention must be paid to molecular bacterial detection on food products subject to freezing or to ionizing irradiation.

Animals↗

Dietary conjugated linoleic acid with fish oil alters yolk n-3 and trans fatty acid content and volatile compounds in raw, cooked, and irradiated eggs.

We investigated the effect of dietary conjugated linoleic acid (CLA) along with n-3 polyunsaturated fatty acid (n-3 PUFA) on yolk fatty acid composition and volatile compounds in eggs that were raw (RA), hard-boiled (HB), or hard-boiled, irradiated (HBI, 2.5 kGy). Single Comb White Leghorn laying hens (n = 40) were randomly assigned to one of the four experimental diets containing 0, 0.5, 1.0, or 2.0% CLA. Menhaden oil was used as the source of n-3 PUFA. Eggs collected after 6 wk of feeding were analyzed for fatty acids and volatile compounds. The content of docosahexaenoic acid (C22:6 n-3) was reduced (P < 0.05) in eggs from hens fed the 2.0% CLA diet. Eggs from hens fed 0.5% CLA incorporated the highest concentration of docosahexaenoic acid (P < 0.05) with a concomitant reduction in arachidonic acid (P < 0.05). The yolk contents of cis-9 trans-11 CLA and trans-10 cis-12 CLA increased linearly (P < 0.05) as the dietary CLA supply increased. Total monounsaturates were reduced (P < 0.05) with an increase in saturates in yolk. No difference was observed in the total PUFA content of eggs. Total volatiles were reduced in RA eggs from 1.0 and 2.0% CLA diets. 2-Propanone, hexane, and methyl cyclopentane were the major volatiles in RA eggs and were reduced by dietary CLA at 1.0 and 2.0%. Acetaldehyde, pentane, propanol, acetic acid methyl ester, acetic acid ethyl ester, propionic acid methyl ester, 2-methylmethyl propionic acid, 2-propanone, and octane were the major volatiles in HB eggs and were reduced by 2.0% CLA (P < 0.05). No difference was observed in the acetaldehyde, pentane, propanol, acetic acid ethyl ester, octane, or total volatile content of HBI eggs.

Animals↗

The effects of milling and processing on wheat contaminated with ochratoxin A.

Samples of sound home-grown wheat (one hard and one soft milling) were obtained, cleaned, and gamma-irradiation used to reduce numbers of viable naturally-occurring fungi. Each sample was inoculated with a toxigenic strain of Penicillium verrucosum and monitored for ochratoxin A formation. When ochratoxin A had reached a level of 60 micrograms/kg, the samples were milled into ten fractions which were analysed for ochratoxin A by an HPLC method with immunoaffinity column clean-up. Each straight-run white flour was baked into bread which was analysed in the same way. Relationships between ochratoxin A levels in naturally-contaminated wheat and the products of milling and baking were established. The recovery of ochratoxin A in wholemeal compared with the cleaned wheat was essentially complete and no significant loss occurred on baking white or wholemeal flour into bread. Recoveries in the straight-run white flours, however, were only approximately one-third for the hard wheat and two-thirds for the soft wheat of the ochratoxin A in the uncleaned wheat. The reason for this was that a much higher proportion of the ochratoxin A was found in the bran and offal fractions from hard wheat than from soft. Conversely, a much higher proportion of the ochratoxin A was found in the reduction flour from soft wheat than from hard. Scouring was examined as a possible method of decontamination of wheat prior to milling. This process removes a proportion of the pericarp (bran coat) prior to milling. The results of the study confirmed that scouring reduced the ochratoxin A level in white and wholemeal flour three-fold for both the hard and soft wheat.

Bread↗

Effect of gamma-radiation and microwave heating on endosperm microstructure in relation to some technological properties of wheat grain.

Grains of the Polish winter wheat variety Begra were subject to gamma-radiation (60Co) within the dose range of 0.05-10 kGy and microwave treatment from 15 to 180 s, the latter resulting in a bulk grain temperature ranging from 28 to 98 degrees C. Both processes were responsible for marked structural changes of wheat kernel endosperm and were pronounced with increasing dose of ionizing radiation or with prolonged time of microwave treatment. Endosperm microstructures of wheat kernels treated with gamma-radiation ranging from 0.05 to 0.5 kGy did not differ from that of untreated ones. Only some changes in the structure of starch granules and proteins were observed at doses of 1, 5 and 10 kGy. Simultaneously at the doses of 5 and 10 kGy a statistically significant decrease of falling number, sodium dodecyl sulfate (SDS) sedimentation values, dough stability and energy were observed, while dough weakening progressed. Microwave treatment longer than 90 s caused marked changes in kernel endosperm structure. Some changes of proteins caused by denaturation created visible fibrils as well as high swelling and deformation of starch granules. This was followed by an increase in the falling number value, decrease in the estimated results of SDS sedimentation test, lowering of the percent of gluten washed out and of the dough energy, respectively. Also a significant decrease in bread quality expressed by volume and score was observed with increasing time of exposure to microwave.

Bread↗