High mutagenic activity formed in pan-broiled pork

Lean pork was pan-broiled at various temperatures between 100 and 290 degrees C. Cooking was performed in an open frying pan common for domestic use in Sweden. No fat was added. Cooking procedures are clearly defined in order to facilitate inter-laboratory comparisons. The crust was extracted with organic solvents of varying polarity. The mutagenic activity was assayed with Ames' Salmonella mutagenicity test. Large amounts of mutagenic activity were detected in samples pan-broiled at 200-290 degrees C. The mutagenic activity recovered was about 10 times higher than that reported by previous investigators to be found during cooking of meat under similar conditions. This discrepancy could be due to differences in the composition of Swedish pork as compared to the meat samples used by other investigators or to different methodology in cooking and extraction procedures.     

Inhibition of Listeria monocytogenes by Lactobacillus bavaricus MN in beef systems at refrigeration temperatures.

The ability of Lactobacillus bavaricus, a meat isolate, to inhibit the growth of three Listeria monocytogenes strains was examined in three beef systems: beef cubes, beef cubes in gravy, and beef cubes in gravy containing glucose. The beef was minimally heat treated, inoculated with L. bavaricus at 10(5) or 10(3) CFU/g and L. monocytogenes at 10(2) CFU/g, vacuum sealed, and stored at 4 or 10 degrees C. The meat samples were monitored for microbial growth, pH, and bacteriocin production. The pathogen was inhibited by L. bavaricus MN. At 4 degrees C, L. monocytogenes was inhibited or killed depending on the initial inoculum level of L. bavaricus. At 10 degrees C, at least a 10-fold reduction of the pathogen occurred, except in the beef without gravy. This system showed a transient inhibition of the pathogen during the first week of storage followed by growth to control levels by the end of the incubation period. Bacteriocin was detected in the samples, and inhibition could not be attributed to acidification. Low refrigeration temperatures significantly (P < or = 0.05) enhanced L. monocytogenes inhibition. Moreover, the addition of glucose-containing gravy and the higher inoculum level of L. bavaricus were significantly (P < or = 0.05) more effective in reducing L. monocytogenes populations in most of the systems studied.     

Mutagenic activity in smoke formed during broiling of lean pork at 200, 250 and 300 degrees C

Smoke formed during pan-broiling of lean pork was recovered at 3 different pan temperatures: 200, 250 and 300 degrees C, using an efficient device for collection of aerosol and volatiles. The mutagenicity of the smoke was assayed using the Ames' Salmonella test. A strong temperature dependence of the mutagen concentration in smoke as well as in meat crust and pan residues was shown. The contribution of mutagenic activity from the smoke relative to the total mutagenicity was 3.1, 4.2 and 11.1% at 200, 250 and 300 degrees C, respectively.     

Mutagenic activity in smoke formed during broiling of lean pork at 200, 250 and 300°C

Smoke formed during pan-broiling of lean pork was recovered at 3 different pan temperatures: 200, 250 and 300°C, using an efficient device for collection of aerosol and volatiles. The mutagenicity of the smoke was assayed using the Ames' Salmonella test. A strong temperature dependence of the mutagen concentration in smoke as well as in meat crust and pan residues was shown. The contribution of mutagenic activity from the smoke relative to the total mutagenicity was 3.1, 4.2 and 11.1% at 200, 250 and 300°C, respectively.     

Effects of nisin and temperature on survival, growth, and enterotoxin production characteristics of psychrotrophic Bacillus cereus in beef gravy.

The presence of psychrotrophic enterotoxigenic Bacillus cereus in ready-to-serve meats and meat products that have not been subjected to sterilization treatment is a public health concern. A study was undertaken to determine the survival, growth, and diarrheal enterotoxin production characteristics of four strains of psychrotrophic B. cereus in brain heart infusion (BHI) broth and beef gravy as affected by temperature and supplementation with nisin. A portion of unheated vegetative cells from 24-h BHI broth cultures was sensitive to nisin as evidenced by an inability to form colonies on BHI agar containing 10 micrograms of nisin/ml. Heat-stressed cells exhibited increased sensitivity to nisin. At concentrations as low as 1 microgram/ml, nisin was lethal to B. cereus, the effect being more pronounced in BHI broth than in beef gravy. The inhibitory effect of nisin (1 microgram/ml) was greater on vegetative cells than on spores inoculated into beef gravy and was more pronounced at 8 degrees C than at 15 degrees C. Nisin, at a concentration of 5 or 50 micrograms/ml, inhibited growth in gravy inoculated with vegetative cells and stored at 8 or 15 degrees C, respectively, for 14 days. Growth of vegetative cells and spores of B. cereus after an initial period of inhibition is attributed to loss of activity of nisin. One of two test strains produced diarrheal enterotoxin in gravy stored at 8 or 15 degrees C within 9 or 3 days, respectively. Enterotoxin production was inhibited in gravy supplemented with 1 microgram of nisin/ml and stored at 8 degrees C for 14 days; 5 micrograms of nisin/ml was required for inhibition at 15 degrees C. Enterotoxin was not detected in gravy in which less than 5.85 log10 CFU of B. cereus/ml had grown. Results indicate that as little as 1 microgram of nisin/ml may be effective in inhibiting or retarding growth of and diarrheal enterotoxin production by vegetative cells and spores of psychrotrophic B. cereus in beef gravy at 8 degrees C, a temperature exceeding that recommended for storage or for most unpasteurized, ready-to-serve meat products.     

Wheat-yellow pumpkin composite flour: Physico-functional,rheological, antioxidant potential and quality properties of pan and flat bread

The impact of yellow pumpkin powder (YP) substitution (5, 10, and 15%) on wheat flour's physico-functional, pasting, gel texture, and dough rheology was studied. Moreover, the nutritional, organoleptic properties and bioactivity of the composite pan and pita bread were evaluated. An improved water holding capacity was noticed for the blended flour than for control. The pasting parameters were declined significantly (p < 0.05) with increasing YP. Composite flours presented a softer gel texture in the presence of YP. Reduced water absorption, increased dough development time, and lower stability for combined flour dough. The bread with YP depicted increased protein, fat, fiber, and mineral contents, while a reduced volume and specific volume were noticed for pan bread. YP incorporated 5% and did not compromise pan and pita bread's color and overall acceptability. Additionally, composite bread depicted higher total phenolics with enhanced antioxidant activities at the higher substitution of YP.     

Mutagenicity,Creatine and Nutrient Contents of Pan Fried Meat from Various Animal Species

The mutagenic activity in extracts of fried meat from 16 different animal species was studied in Salmonella typhimurium TA98. In each experiment, 1 meat sample together with a standard beef sample was fried, and the mutagenicity was expressed relative to the beef sample. All meat samples showed less mutagenic activity than beef. The contents of creatine, creatinine, water, protein, carbohydrate and fat in the meat samples were analyzed, but mutagenicity was not correlated with the concentration of any of these constituents. Beef meat treated with creatinase to remove creatine produced reduced mutagenic activity. Possibly a threshold concentration of creatine is necessary to give a high mutagenic response.     

Influence of frying fat on mutagenic activity in lean pork meat

Mutagenic activity in lean pork meat fried at two different pan temperatures, 200 degrees C and 250 degrees C, with or without the addition of fat, was measured in Ames' Salmonella test on strain TA98. 9 different fats with varying chemical composition were tested. All fried meat samples were shown to be mutagenic. At the frying temperature of 200 degrees C differences between meat samples fried in different fats or without fat, respectively, were small. All meat samples fried at 250 degrees C were considerably more mutagenic than the samples fried at 200 degrees C. At 250 degrees C, the addition of fat caused a significant rise in mutagenic activity. We believe this is mainly an effect of more efficient heat transfer from the bottom of the frying-pan to the meat samples, although other factors may also contribute.     

Mutagenic properties of 2-amino-N6-hydroxyadenine in Salmonella and in Chinese hamster lung cells in culture

The mutagenicity of the base analogue, 2-amino-N6-hydroxyadenine (AHA), was tested in Salmonella typhimurium TA100 and TA98 and in Chinese hamster lung (CHL) cells. AHA showed very potent mutagenicity in TA100 without S9 mix, inducing 25,000 revertants/micrograms. The mutagenicity increased about 2-fold upon addition of S9 mix containing 10 microliters S9. AHA was found to be one of the strongest mutagens for TA100. Addition of S9 mix containing 100 microliters S9 induced no significant increase of revertants with AHA at amounts up to 50 ng per plate. AHA was also mutagenic for the frameshift mutant, TA98, without S9 mix, the mutagenicity for TA98 being about 1/1000 of that for TA100. When the mutagenicity of AHA was tested in CHL cells, with diphtheria toxin resistance (DTr) as a selective marker in the absence of S9 mix with a 3-h treatment of cells, DTr mutants increased dose-dependently at concentrations of 2.5-15 micrograms/ml. When cells were incubated with AHA for 24 h, a 200-fold increase in the number of DTr mutants was observed; the mutagenicity was 500-fold higher than that of ethyl methanesulfonate. This marked increase of mutagenicity by prolonged incubation may indicate that AHA induces mutations mainly after incorporation into DNA. The addition of a small amount of S9 increased the mutagenicity obtained with a 3-h treatment 2-fold, but a larger amount of S9 decreased the mutagenicity as was found with S. typhimurium TA100.     

Fecal mutagenicity arising from ingestion of fried ground beef in the human

Fried ground beef has been shown to contain mutagens, and the major mutagenic component has been identified as 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx). Mutagens in feces of 3 adult volunteers were fractionated by treatment of the feces with blue cotton followed by chromatography on a carboxymethyl cellulose column. The chromatographic fraction, corresponding to MeIQx in terms of the position of elution, was examined for mutagenicity in S. typhimurium TA98 with metabolic activation. When meals containing no heated meat were eaten, this fraction of feces showed little or no mutagenicity. On eating fried ground beef, the feces excreted in the next two days showed greatly increased mutagenicity in this fraction. By eating no-meat meal subsequent to the meat meal, the mutagenicity resumed the original low level on the fourth day after the meat meal. The components in the mutagenic fraction were probably metabolites of the mutagens present in cooked meat, since analysis by high pressure liquid chromatography of the mutagenic fraction showed that the active components in the feces were different from the mutagens in cooked meat.     

Influence of fried meat and fiber on cytochrome P-450 mediated activity and excretion of mutagens in rats

Fried meat was included in the diet of Sprague-Dawley rats during one week. Ingested and excreted amounts of mutagenic activity were determined daily by the use of the Ames' Salmonella/mammalian microsome test on extracts of the diet, urine and feces. In addition, the effect of the fried meat on ethoxyresorufin-O-deethylase activity in the small intestine and in the liver was measured. The results were compared to those from a control group of rats receiving boiled instead of fried meat as their source of protein. The diet containing boiled meat was not mutagenic and none of the samples from the control group, neither urine nor feces, contained any mutagenicity. The activity of ethoxyresorufin O-deethylase in the small intestine was increased by fried meat whereas this enzyme activity in the liver was unaffected. O-deethylation of ethoxyresorufin is catalyzed by cytochrome P-450 dependent enzyme(s) and induction of the enzyme activity might indicate an increased metabolic activation of premutagens and precarcinogens in the intestine. It is not yet known whether the mutagens excreted by the animals receiving the fried meat diet represent unmetabolized dietary mutagens or are formed as a result of metabolic activation of compounds in fried food. Dietary fibers i.e. wheat bran, pectin, cellulose or ViSiblin were included in the above-described diets and were shown to lower the mutagenic activity in urine and feces of the rats. Pectin significantly increased the hepatic ethoxyresorufin-O-deethylase and decreased the 16 alpha-hydroxylation of 4-androstene-3,17-dione.     

Mutagen formation in fried meat emulsion containing various amounts of creatine

The formation of mutagens in fried, minced meat emulsion was evaluated by the Ames Salmonella test system. Exogenous addition of creatine to the emulsion prior to frying greatly enhanced the mutagenicity of the emulsion. Addition of 5% creatine resulted in a 40-fold increase in the mutagenicity of the fried meat emulsion in the frameshift test strain, TA98, and in a 8-fold increase in the base substitution test strain, TA100. The present results suggest that creatine is an important factor in the mutagen formation in fried meat products.     

Mutagenic and carcinogenic heterocyclic amines in Chinese cooked foods

Samples of 7 foods commonly eaten in the Northeast of China (i.e. fried and broiled fishes and broiled meat) were tested for mutagenicity on Salmonella typhimurium TA98 with S9 mix. The basic fractions of the samples were mutagenic, inducing 33-2930 revertants/g of cooked food. Fried walleye pollack (a kind of cod fish heated on a stainless steel pan) showed the highest mutagenicity, so attempts were made to isolate mutagens from the basic fraction of this food. The mutagens were purified by treatment with blue cotton and HPLC on a semi-preparative ODS column and analytical cation exchange and ODS columns. 5 mutagens were isolated and identified as 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). 1 g of fried fish was estimated to contain 0.16 ng of IQ, 0.03 ng of MeIQ, 6.44 ng of MeIQx, 0.10 ng of 4,8-DiMeIQx and 69.2 ng of PhIP. MeIQx and PhIP accounted for 24% and 4.7%, respectively, of the total mutagenicity. The other 3 heterocyclic amines were each responsible for only 0.3-1.2% of the total mutagenicity.     

Meat-derived carcinogens,genetic susceptibility and colorectal adenoma risk

Exposure to heterocyclic aromatic amines (HAAs), carcinogens produced when meat is cooked at high temperatures, is an emerging risk factor for colorectal cancer (CRC). In a cross-sectional study of 342 patients undergoing a screening colonoscopy, the role of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (DiMeIQx), the three most abundant HAAs found in cooked meats, and total mutagenic activity in cooked meats were examined in relation to colorectal adenoma risk. Given that genetic differences in the ability to biotransform HAAs and repair DNA are postulated to modify the HAA–CRC relationship, gene–diet interactions were also examined. Among the total study population, no relationships were observed between dietary HAAs or meat mutagenicity, and colorectal adenoma risk; however, in males, positive associations between dietary HAAs/meat mutagenicity exposures and adenoma risk were suggestive of a relationship. In a separate analysis, polymorphisms in CYP1B1 were found to be associated with colorectal adenoma risk. Additionally, gene–diet interactions were observed for dietary PhIP and polymorphisms in CYP1B1 and XPD, dietary DiMeIQx and XPD polymorphisms, and meat mutagenicity exposure and CYP1B1 polymorphisms. Overall, increased colorectal adenoma risk was observed with higher HAA/meat mutagenicity exposures among those with polymorphisms which confer greater activity to biotransform HAAs and/or lower ability to repair DNA. This research supports the link between dietary HAAs and genetic susceptibility in colorectal adenoma etiology. The vast majority of CRCs arise from colorectal adenomas; thus, the results of this study suggest that changes in meat preparation practices limiting the production of HAAs may be beneficial for CRC prevention.     

Presence of nitrosable mutagen precursors in cooked meat and fish

Broiled chicken, pork, mutton, beef and sun-dried sardine were found to yield direct-acting mutagenicity after nitrite treatment. When 50% methanol extracts of cooked foods were treated with 50 mM nitrite at pH 3 for 1 h at 37 degrees C, they induced 3800-17,900 revertants of Salmonella typhimurium TA100 and 15,000-43,600 revertants of TA98 per g. In contrast, raw meat and uncooked sun-dried sardine showed little or no mutagenicity after nitrite treatment. Treatment of broiled chicken with 0.5-3 mM nitrite, which is a physiologically feasible concentration in the human stomach under some conditions, induced direct-acting mutagenicity. When broiled chicken was treated with 1 mM nitrite at pH 3 for 1 h at 37 degrees C, its mutagenicities on TA100 and TA98 without S9 mix were 7100 and 5400 revertants/g, respectively.     

Bacteriological Survey of Frozen Meat and Gravy Produced at Establishments Under Federal Inspection

During visits to 34 federally inspected establishments producing frozen meat and gravy, 541 production line samples and 535 finished product units were collected for bacteriological analyses. It was found that more than 70% of the sets of finished product (10 units/set) produced under good manufacturing practices had: (i) four or fewer coliform-positive units, (ii) two or fewer Escherichia coli-positive units, (iii) three or fewer Staphylococcus aureus-positive units, and (iv) an aerobic plate count of fewer than 50,000/g (geometric mean of 10 units). All finished product units were negative for salmonellae.     

Bacteriology of Dehydrated Space Foods

The initial bacteriological requirement established in 1964 for space foods by the U.S. Army Natick Laboratories are: a total aerobic plate count ( 300,000), chocolate ice cream cubes (20,000), and each of four samples of chocolate candy (12,000 to 61,000); (ii) coliforms: two out of three vanilla milk drinks (16 and 127) and one beef hash bar (14); (iii) fecal coliforms: one sample of chicken soup and gravy base positive; (iv) fecal streptococci: two samples of peanut cubes (40 and 108), coconut cubes (75), chicken soup and gravy base (2,650), beef soup and gravy base (33), and five out of six flavored milk drinks (23 to 300); (v) salmonellae: one each of chicken and beef soup and gravy base were positive.     

Growth and bone mineralisation as affected by dietary calcium, phytic acid and vitamin D

1. Rats were fed various diets ranging from the normal chow, pure flour containing large amounts of phytic acid, Ca-enriched flour and mixtures of flour and normal food with various levels of calcium. 2. It was found that the animals eating the pure flour grew less and were smaller. 3. They suffered from hypocalcemia and had low plasma alkaline phosphatase and 25-HCC-vitamin D3 levels. 4. These animals had rib-cage deformities. 5. Additional calcium in the flour improved the animals' growth and calcification. 6. The mixed food did not greatly affect the animals and additional calcium did not improve growth or bone mineralisation. 7. The Bedouin eat large amounts of unleavened bread containing large amounts of phytates. 8. It is concluded that uptake of large amounts of phytates by the Bedouin eating unleavened bread is due to the flour and that the clinical manifestations are a direct result of the flour and not the lack of vitamin D due to covering the skin from sunlight.     

Hydrophobicity of stored (15, 35 °C), or dry-heated (120 °C) rice flour and deteriorated breadmaking properties baked with these treated rice flour/fresh gluten flour

Rice flour was stored at 15 °C/9 months, at 35 °C/14 days, or dry-heated at 120 °C/20 min. The breadmaking properties baked with this rice flour/fresh gluten flour deteriorated. In addition, the rice flour was mixed with oil in water vigorously, and oil-binding ability was measured. Every rice flour subjected to storage or dry-heated at 120 °C showed higher hydrophobicity, owing to changes in proteins. Then, proteins in the stored rice flour were excluded with NaOH solution, and bread baked with the deproteinized rice flour showed the same breadmaking properties as unstored rice flour/fresh gluten flour. The viscoelasticity of wheat glutenin fraction decreased after the addition of dry-heated rice flour in a mixograph profile. DDD staining increased Lab in color meter, which suggested an increase in SH groups in rice protein. The increase in SH groups caused a reduction in wheat gluten protein resulting in a deterioration of rice bread quality.