Subacute feeding toxicity of low-sodium sausages manufactured with sodium substitutes and biopolymer-encapsulated saltwort (Salicornia herbacea) in a mouse model.

BACKGROUND: Low-sodium sausages were manufactured using sodium substitution and biopolymer encapsulation. A diet comprising 10% treatment sausages (six treatment groups: C (100% NaCl), T1 (55% sodium substitute + 45% saltwort salt), T2 (55% sodium substitute + 45% saltwort salt with chitosan), T3 (55% sodium substitute + 45% saltwort salt with cellulose), T4 (55% sodium substitute + 45% saltwort salt with dextrin), and T5 (55% sodium substitute + 45% saltwort salt with pectin)) was added to a 90% commercial mouse diet for 4 weeks. RESULTS: Subacute toxicity, hematology, liver function, and organ weight tests in low-sodium sausage groups showed results similar to those of the control group, and all toxicity test levels were within normal ranges. CONCLUSIONS: All low-sodium sausage types tested are suggested to be safe in terms of subacute toxicity. Moreover, low-sodium sausages can be manufactured by biopolymer encapsulation of saltwort using pectin, chitosan, cellulose, and dextrin without toxicity. © 2019 Society of Chemical Industry.

Induction of colonic aberrant crypts in mice by feeding apparent N-nitroso compounds derived from hot dogs.

Nitrite-preserved meats (e.g., hot dogs) may help cause colon cancer because they contain N-nitroso compounds. We tested whether purified hot-dog-derived total apparent N-nitroso compounds (ANC) could induce colonic aberrant crypts, which are putative precursors of colon cancer. We purified ANC precursors in hot dogs and nitrosated them to produce ANC. In preliminary tests, CF1 mice received 1 or 3 i.p. injections of 5 mg azoxymethane (AOM)/kg. In Experiments 1 and 2, female A/J mice received ANC in diet. In Experiment 1, ANC dose initially dropped sharply because the ANC precursors had mostly decomposed but, later in Experiment 1 and throughout Experiment 2, ANC remained at 85 nmol/g diet. Mice were killed after 8 (AOM tests) or 17-34 (ANC tests) wk. Median numbers of aberrant crypts in the distal 2 cm of the colon for 1 and 3 AOM injections, CF1 controls, ANC (Experiment 1), ANC (Experiment 2),and untreated A/J mice were 31, 74, 12, 20, 12, and 5-6, with P < 0.01 for both ANC tests. Experiment 2 showed somewhat increased numbers of colonic mucin-depleted foci in the ANC-treated group. We conclude that hot-dog-derived ANC induced significant numbers of aberrant crypts in the mouse colon.

Consumers' understanding and concerns about bovine spongiform encephalopathy (BSE): comparison among Canadian, American, and Japanese consumers.

In spite of much analysis of the impact of bovine spongiform encephalopathy (BSE) on consumer perceptions and meat purchases, there has been little explicit analysis of the level of BSE knowledge. In this study the role of knowledge about BSE was examined in Canada, the United States, and Japan. In addition, the level of knowledge was linked to human health concerns regarding BSE and whether there is agreement with paying a premium for beef with BSE animal tests. From a public policy perspective, understanding whether higher or lower knowledge is linked to public concern and desire for market intervention might help in the design of risk communication in any future animal disease outbreak. Should lack of knowledge about the disease be related to a public desire for market intervention (animal testing, for example), then an increase in detailed information about how humans might contract the disease might change public pressure for intervention. As compared to U.S. and Canadian respondents, Japanese respondents are more knowledgeable regarding the ways in which humans might be exposed to the human variant of BSE (variant Creutzfeldt-Jakob disease, vCJD) and are more concerned about the disease. However, U.S. respondents are more willing to pay a premium for beef tested to ensure that it will not result in vCJD. Japanese respondents who are more knowledgeable about BSE are more concerned about the risk of BSE to human health. In Canada, subjects who are more knowledgeable about the ways in which humans attain vCJD are less concerned about the risk of BSE to human health. Knowledge of the ways in which humans develop vCJD does not significantly influence concerns about the risk of BSE to human health in the United States or willingness to pay for BSE-tested beef in any of the three countries. The links between knowledge and concerns about BSE and between knowledge and agreement with paying premiums for BSE-tested beef were estimated for each country using ordered probit regressions.

Public risk perception of relaxation of transmissible spongiform encephalopathies (TSE) measures in Europe.

The so-called "TSE roadmap" was published by the European Commission on July 15, 2005. The transmissible spongiform encephalopathy (TSE) roadmap suggests relaxation of bovine spongiform encephalopathy (BSE) in cattle and other animal transmissible spongiform encephalopathies measures in the short, medium, and long term. According to the TSE roadmap, "Any relaxation of BSE measures following the scientific assessment should be initiated by an open discussion with all stakeholders and supported by a strong communication strategy" ( European Commission 2005 , 5). Bearing this in mind, a social scientific project was designed to (1) involve different stakeholder groups, governmental risk managers, and their scientific advisors and (2) obtain their perception of the TSE roadmap and of its implications for precautionary consumer protection in five European Union (EU) Member States. This study describes the risk perception and risk management of TSE in Europe as exemplified by the TSE roadmap. The following query guided the international comparative study: How is TSE risk perceived by four interviewed stakeholder groups in five studied countries? The risk perceptions of TSE of risk managers from the ministries in charge in Belgium, France, Germany, Italy, and the United Kingdom, as well as their scientific advisors and stakeholder groups, were determined. The stakeholder groups were from three different areas involved with TSE, including farmers, consumers, and the meat/food industry. The issue to be addressed is roadmapping an adequate instrument for stakeholder involvement and for risk decision making.

Enhanced fat consumption potentiates acrylamide-induced oxidative stress in epididymis and epididymal sperm and effect spermatogenesis in mice.

Acrylamide (ACR) and high contents of fat could be found co-existent in many foods processed by high temperature, such as deep-frying and roasting. This study investigated the effect of enhanced fat consumption on deficits of spermatogenesis induced by ACR, and explored potential mechanisms of oxidative damage involved in this pathology in mice. Results show that enhanced feeding of corn oil and pork fat on mice potentiated the decreases of spermatogonia along with mature sperms after treatment of ACR, and that spermatozoa quality is significantly reduced as a result of enhanced feeding of corn oil and pork fat on mice treated with ACR. Moreover, enhanced consumption of corn oil and pork fat potentiated the up-regulation of malondialdehyde (MDA) level in epididymal sperm and cauda epididymides, also up-regulated level of Protein carbonyls (PCOs) in cauda epididymides, of mice after treatment of ACR. Last, enhanced consumption of corn oil and pork fat potentiated the reduced activity of superoxide dismutases (SOD) in epididymal sperm, corpus, and cauda epididymides, also reduced activity of glutathione peroxidase (GPx) in cauda epididymides, of mice treated with ACR. These data suggest that enhanced feeding of corn oil and pork fat on mice potentiates ACR-induced oxidative stress in the epididymis and epididymal sperm and a subsequent effect on spermatogenesis.

Processed meat intake and chronic disease morbidity and mortality: An overview of systematic reviews and meta-analyses.

Despite the nutritional value of meat, a large volume of reviews and meta-analyses suggests that processed meat intake is associated with an increased risk of chronic diseases. However, assessments of the quality of these published reviews internal validity are generally lacking. We systematically reviewed and assessed the quality alongside summarizing the results of previously published systematic reviews and meta-analyses that examined the association between processed meat intake and cancers, type II diabetes (T2D), and cardiovascular diseases (CVD). Reviews and meta-analyses published until May 2018 were identified through a systematic literature search in the databases MEDLINE and EMBASE, and reference lists of included reviews. The quality of the systematic reviews and meta-analyses was assessed using A Measurement Tool to Assess Systematic Reviews (AMSTAR). All eligible reviews had to comply with two quality requirements: providing sufficient information on quality assessment of the primary studies and a comprehensive search. The results were summarized for T2D, CVD, and each of the different cancer types. The certainty in the estimates of the individual outcomes was rated using the Grading of Recommendations, Assessment, Development and Evaluations (GRADE) method. In total, 22 systematic reviews were eligible and thus included in this review. More than 100 reviews were excluded because quality assessment of the primary studies had not been performed. The AMSTAR score of the included reviews ranged from 5 to 8 indicating moderate quality. Overall, the quality assessments of primary studies of the reviews are generally lacking; the scientific quality of the systematic reviews reporting positive associations between processed meat intake and risk of various cancers, T2D and CVD is moderate, and the results from case-control studies suggest more often a positive association than the results from cohort studies. The overall certainty in the evidence was very low across all individual outcomes, due to serious risk of bias and imprecision.

Beneficial effect of an omega-6 PUFA-rich diet in non-steroidal anti-inflammatory drug-induced mucosal damage in the murine small intestine.

AIM: To investigate the effect of a fat rich diet on non-steroidal anti-inflammatory drug (NSAID)-induced mucosal damage in the murine small intestine. METHODS: C57BL6 mice were fed 4 types of diets with or without indomethacin. One group was fed standard laboratory chow. The other groups were fed a fat diet consisting of 8% w/w fat, beef tallow (rich in SFA), fish oil, (rich in omega-3 PUFA), or safflower oil (rich in omega-6 PUFA). Indomethacin (3 mg/kg) was injected intraperitoneally from day 8 to day 10. On day 11, intestines and adhesions to submucosal microvessels were examined. RESULTS: In the indomethacin-treated groups, mucosal damage was exacerbated by diets containing beef tallow and fish oil, and was accompanied by leukocyte infiltration (P < 0.05). The mucosal damage induced by indomethacin was significantly lower in mice fed the safflower oil diet than in mice fed the beef tallow or fish oil diet (P < 0.05). Indomethacin increased monocyte and platelet migration to the intestinal mucosa, whereas safflower oil significantly decreased monocyte and platelet recruitment (P < 0.05). CONCLUSION: A diet rich in SFA and omega-3 PUFA exacerbated NSAID-induced small intestinal damage via increased leukocyte infiltration. Importantly, a diet rich in omega-6-PUFA did not aggravate inflammation as monocyte migration was blocked.

Comparison of the mutagenic activity of various brands of food grade beef extracts.

The mutagenic activity of 9 brands of commercial meat extracts were compared using Ames tester strain TA 98 with hepatic S-9 activation prior to concentration by solvent extraction. In a preincubation test system where 20 mg of intact meat extract preparations were tested per plate, about half of the brands were non-mutagenic whereas the other half increased the spontaneous revertant number by a factor of 2.5-3 (MF). The mutagenic principals could be extracted into methylene chloride under alkaline conditions and the mutagenic activity of these basic organic extracts ranged in potency from a mutation factor of 7-40 (280-1600 revertants) per 2 g equivalents of intact meat extracts in the standard Ames test. A quantitative correlation was obtained between the two trials (intact and extracted samples). The commercial meat extract with the greatest activity contained approximately 70 ng/g of total amino-imidazoquinoline and amino-imidazoquinoxaline compounds while the weakest brand of meat extract contained less than 10 ng/g of these heterocyclic aromatic amines. The human intake of these heterocyclic amines from meat extract in bouillon soups was estimated at maximally 100 ng/day and is approximately 10(3) times less than the estimated total daily intake of all heterocyclic amines which are known to be formed in protein-rich heated foods.

Urinary mutagens in humans after fried pork and bacon meals.

Urinary mutagenic activity on Salmonella typhimurium strain TA1538 with S9 was determined after morning meals of fried pork and bacon. Both in fasting and non-fasting subjects a very marginal elevation of urinary mutagenic activity was observed, accounting for a small fraction only of the total amount of mutagens ingested.

Mutagenic activity in the stomach and small intestine of rats after oral administration of nitrosated beef extract.

The products formed by the reaction of beef extract with nitrite were assayed in the Salmonella/microsome mutagenicity test. In strain TA1538, TA98 and TA100 a direct-acting mutagenic response was observed. The presence of liver-microsome preparation resulted in decreased mutagenicity. To study the absorption, distribution and excretion of mutagenic substances in nitrosated beef extract, the test material was given perorally to rats. Investigations of the stomach, bile fluid, urine, small intestine and blood samples were carried out, and mutagenicity was found in the contents of stomach and small intestine. It is supposed that unlike beef extract itself, its nitroso product is not excreted in the bile but passes directly from the stomach and small intestine.

Inhibition by quercetin and luteolin of chromosomal alterations induced by salted, deep-fried fish and mutton in rats.

Previous studies from our laboratory have shown the clastogenic effects of long-term feeding on deep-fried fish and mutton in rat bone marrow cells. We report the chemopreventive action of two flavanoids, quercetin (Qn) and luteolin (Ln) against the induced mutagenicity by fish and mutton extracts. Groups of rats were treated with flavanoids through pre-, simultaneous- and post-treatment regimens and killed at the end of treatment. The bone marrow was removed and analysed for the presence of micronuclei (MN) and chromosome aberrations (CA). Pre-treatment showed most effectively a good inhibition of mutagenicity at every dose tested. Luteolin was a better protective agent than quercetin. It protected the cells against genetic damage to 93% in the micronucleus assay and to 95% in the chromosome aberrations induced by fish extract (p < 0.001 in both the groups). Mutton extract-induced micronuclei and chromosome aberrations were protected 85% and 90%, respectively, by luteolin and 79% and 76%, respectively, by quercetin. Our results tend to suggest that quercetin and luteolin are potential chemopreventive compounds.

Mutagenicity of cooked hamburger is reduced by addition of onion to ground beef.

Addition of onion effectively reduced mutagenicity of cooked hamburger when tested on Salmonella typhimurium TA98 strain with metabolic activation. The components of onion that participated in the reduction of mutagenicity were sugars. Addition of starch or glucose to ground beef the amount equivalent to that in onion reduced the mutagenicity of cooked hamburger. Addition of onion may cause imbalance of the sugar content of ground beef that effectively produces mutagenicity. Mutagenicity of the heated model mixture of glucose/glycine/creatinine in diethyleneglycol-water was reduced by an excessive amount of glucose. Hence, Japanese cooking-style with addition of onion can reduce mutagenicity of hamburger.

Mutagenicity of cooked hamburger is controlled delicately by reducing sugar content in ground beef.

Effect of sugars added to ground beef on the generation of mutagenicity of cooked hamburger was investigated. Mutagenicity of hamburger was assayed by the Ames test using Salmonella typhimurium TA98 strain with metabolic activation after the mutagens were purified by use of blue rayon. Intrinsic reducing sugar content in ground beef was estimated to be 0.07% (w/w). Mutagenicity of hamburger was sharply or delicately controlled by the amount of a reducing sugar added to ground beef. Mutagenicity was increased more than 2-folds by addition of 0.08% (w/w) glucose, fructose or lactose but decreased to about a half by addition of more than 0.67% (w/w) each of the reducing sugars. Mutagenicity of cooked hamburger was not influenced by addition of sucrose at the ranges between 0.08 and 0.67% (w/w). When red wine with 0.10% (w/w) equivalent amount of reducing sugars or white wine with 0.13% (w/w) equivalent amount of reducing sugars were added to the ground beef, mutagenicity of cooked hamburger was similarly increased 1.6-1.8-fold. Controlling the reducing sugar content in ground beef would be a simple way to regulate the mutagenicity of cooked hamburgers.

Nitrite-cured meats as a source of endogenously and exogenously formed N-nitrosoproline in the ferret.

Nitrite-cured meat containing 120 mg Na15NO2/kg was fed to male ferrets (Mustela putorius furo). During consumption of the meat, the animals were dosed orally with 0.87 mmol [2-2H]proline. All urine was collected throughout the study and analysed for total N-nitrosoproline (NPRO) and isotopic enrichment of NPRO by mass spectrometry. The cured-meat diet increased the urinary excretion of NPRO 14-fold. Isotope analyses indicated that approximately 70% of the NPRO came from the cured meat, the majority of which was analytically unavailable or 'bound' NPRO in the meat. A small portion of the excreted NPRO appeared to be formed in the stomach as a result of ingesting the cured meat. A minor amount of the excreted NPRO did not contain any isotopically labelled atoms. The administration of ascorbic acid did not significantly alter NPRO excretion. Animals dosed orally with 11.4 mumol of a peptide in which the N-terminal proline was nitrosated increased their excretion of NPRO by 385 nmol over the following 48 hr. These data indicate that nitrite-cured meat contains bound NPRO which contributes to the total amount of NPRO in the urine.

Effects of creatine and creatinine content on the mutagenic activity of meat extracts, bouillons and gravies from different sources.

Thirteen commercial meat-flavour samples were analysed for creatine and creatinine content and tested for mutagenicity in the Ames Salmonella/microsome test. In most samples, more than 50% of the creatine had been converted to creatinine. Mutagenicity was related to the creatinine content: 150 mumol creatinine/g dry matter (gdm) gave upwards of 2500 revertants/gdm, concentrations of 15-40 mumol/gdm gave about 100 revertants/gdm and concentrations of 1-10 mumol/gdm gave only low or no significant mutagenicity. No relationship was apparent between coloration and mutagenicity. Beef steaks (initial weight c. 500 g) baked at oven temperatures between 115 and 245 degrees C only showed significant mutagenicity--135 revertants/100 gE (initial raw weight)--in the crust when baked at the highest temperature (245 degrees C). The gravies (meat-juice drip) collected during baking showed a linear increase in mutagenicity with baking temperatures up to 180 degrees C (48-828 revertants/100 gE) and a very sharp increase in mutagenicity for the gravy collected from beef steak baked at 245 degrees C (28,300 revertants/gdm or 19,800 revertants/100 gE). At this high temperature, the brown coloration and the proportion of creatinine to total creatine and creatinine were also dramatically increased, because this gravy dried up completely during the baking procedure.

Analysis of commercial bouillons for trace levels of mutagens.

A new method, developed specifically for the extraction of heterocyclic aromatic amine (HAA) type mutagens from different food matrices, was applied to various forms of commercially available bouillons. This procedure is based on liquid-liquid extraction of the sample at different pH values. Recovery and reproducibility of the procedure was determined by processing spiked samples using a mutagenicity bioassay technique as an endpoint. The mutagenicity was tested in the Salmonella/microsome assay using strain TA98 with metabolic activation. 22 bouillon samples in liquid, cube or powder forms from seven manufacturers were extracted and tested for potential mutagenicity. The mutagenic activity of these samples varied and ranged from non-detectable to about 1200 induced revertants per gram of solid material, with a median value of approximately 250 revertants/g. The mutagenic response appeared to be dependent on the source rather than the type or form of the product tested. A negative response was obtained from only one chicken bouillon, and the highest positive response was obtained from a beef bouillon in cube form. It appears that the average beef sample, regardless of form, has a higher mutagenic potency than chicken or chicken and turkey samples. Overall, the intake of mutagens from commercial bouillons (obtained as cubes, concentrates or dry mixes) to prepare one serving (as bouillon, soup, casseroles, etc.) is considerably less than that reported in the literature for one serving of fried beef or pork. The extractability and mutagenic characteristics of these samples indicate the presence of HAA-type mutagens. Work is in progress to identify the mutagenic factors in bouillons.

Formation of direct-acting genotoxic substances in nitrosated smoked fish and meat products: identification of simple phenolic precursors and phenyldiazonium ions as reactive products.

Epidemiological studies have associated the consumption of smoked fish and meat products with an increased risk of stomach cancer. Therefore, the reaction of such smoked foods with nitrite under acidic conditions was investigated and was shown to produce potent direct-acting genotoxic substances as detected by the SOS Chromotest. Similar genotoxic activity was observed in nitrosated samples of wood-smoke condensates. Simple phenolic compounds such as phenol, 3-methoxycatechol, catechol and vanillin were identified as the precursors of the genotoxic substances. These phenolic compounds also exhibited direct-acting genotoxicity after nitrosation. The major genotoxic substances formed after nitrosation of phenol were isolated and identified as 4- and 2-hydroxyphenyldiazonium ions. Nitrosation of various wood-smoke condensates was found to generate the same type of diazonium compounds, which in part account for the genotoxicity of nitrosated smoked foods.

Genotoxicity testing of cooked cured meat pigment (CCMP) and meat emulsion coagulates prepared with CCMP.

The preformed cooked cured meat pigment (CCMP) synthesized directly from bovine red blood cells or through a hemin intermediate was found to be a viable colorant for application to comminuted pork as a nitrite substitute. However the genotoxicity of CCMP and meat emulsion coagulates prepared with CCMP has not been evaluated. Therefore the objectives of this work were to investigate genotoxicity of CCMP and the influence of CCMP addition on genotoxicity and the content of residual nitrite in model meat emulsion coagulates. Meat emulsions were prepared from white (musculus longissimus dorsi) and red (musculus quadriceps femoris) pork muscles with two different amounts of synthesized pigment CCMP. Comparatively, emulsions with fixed addition of nitrite salt and emulsions without any addition for color development were made. Genotoxicity of CCMP and meat emulsion coagulates was tested with the SOS/umu test and the Ames test. Neither CCMP nor meat emulsion coagulates prepared with CCMP or nitrite salt were genotoxic in the SOS/umu test. In the Ames test using Salmonella Typhimurium strains TA98 and TA100 samples of coagulates prepared with CCMP and with nitrite showed weak mutagenic activity in Salmonella Typhimurium strain TA100 but only in the absence of the metabolic activation, while CCMP was not mutagenic. Coagulates prepared with CCMP contained significantly less residual nitrite than coagulates prepared with nitrite salt. These results indicate that from the human health standpoint the substitution of nitrite salt with CCMP would be highly recommendable.

Validation of ELISA test kits for detection of beta-agonist residues in meat.

Three ELISA test kits, the Randox ELISA beta-agonist test kit, Euro-Diagnostica test kit, and Ridascreen beta-agonist test kit, were evaluated for screening of meat and liver for beta-agonist residues in fortified and field-incurred samples. It was found that the Randox beta-agonist test kit was more suitable as a screening tool due to its accuracy, ease of use, and lower cost. The tests were able to detect beta-agonist residues at the minimum level of detection, as claimed by the suppliers. The performance of the method as assessed through recovery rates of beta-agonists in fortified samples was satisfactory with a low coefficient of variation (1-3%). Repeatability, as measured through the coefficient of correlation was also satisfactory. For field-incurred positive samples, the test kit showed a sensitivity of 100% and a low rate of false positives for goat and cow tissues. However, a high rate of apparent false positives was obtained for tissues of swine.

Determination of six illegal antibiotics in chicken jerky dog treats.

In 2007 chicken jerky dog treats were implicated in causing illnesses and death in dogs in several countries. Affected dogs were diagnosed with acquired Fanconi syndrome, which is characterized by kidney malfunction. Known causes of this condition include a chemical assault by various contaminants including certain drugs. For this reason investigations into possible causes of the illnesses included antibiotics that may be used in animal husbandry. Targeted analyte screens of individual imported chicken jerky dog treats using LC-MS/MS detected six illegal antibiotics in imported products of several brands. Trimethoprim, tilmicosin, enrofloxacin, sulfaclozine, and sulfamethoxazole are not allowed in chicken at any level and were found as high as 2800 ng/g (ppb). Sulfaquinoxaline was found in chicken jerky treats as high as 800 ng/g, which is well above the U.S. FDA tolerance of 100 ng/g. Although there is no evidence these contaminants were responsible for the dog illnesses, their misuse could contribute to antibiotic-resistant strains of bacteria.