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Production of Bacillus cereus emetic toxin (cereulide) in various foods.

To determine the role of Bacillus cereus as a potential pathogen in food poisoning, the production of an emetic toxin (cereulide) by B. cereus was quantified in various food sources. The amount of emetic toxin in 13 of 14 food samples implicated in vomiting-type food poisoning cases ranged from 0.01 to 1.28 microg/g. A vomiting-type strain, B. cereus NC7401, was inoculated into various foods and incubated for 24 h at 20, 30, and 35 degrees C. In boiled rice, B. cereus rapidly increased to 10(7)-10(8) cfu/g and produced emetic toxin at both 30 and 35 degrees C. In farinaceous foods, the production of emetic toxin was as high as that in the food samples implicated in food poisoning. Low levels of emetic toxin were detectable in egg and meat and their products and a small quantity of toxin was detectable in liquid foods such as milk and soymilk when not aerated. Bacterial growth and toxin production was inhibited in foods cooked with vinegar, mayonnaise, and catsup, supposedly by the decreased pH of acetic acid. This is the first report that has quantified emetic toxin of B. cereus in various foods.

Bacillus cereus↗

Estimates of mineral intakes using food composition tables vs measures by inductively-coupled plasma mass spectrometry: Part 1. calcium, phosphorus and iron.

OBJECTIVES: To examine the accuracy of food composition table (FCT)-based estimation of dietary nutrient element intake in reference to the instrumental measurement by inductively-coupled plasma mass spectrometry (ICP-MS). DESIGN: Cross-sectional study. SETTING: Communities. SUBJECTS: 884 adult non-smoking women in 23 study sites in 6 areas in Asia (i.e. China (Mainland), China (Taiwan), Korea, Japan, Malaysia and the Philippines. METHODS: 24-hour food duplicate samples foods offered by the women were subjected (1) to the estimation of daily dietary intake of calcium (Ca), phosphorus (P) and iron (Fe), using 6 types of national FCTs one for each area, and (2) to ICP-MS analysis for the measurement of the same elements. RESULTS: Comparison of the estimated values (E) with corresponding measured values (M) in terms of the mean (E)/(M) ratio for each area showed that the ratio was various among the 6 areas studied. Ca showed smallest deviation (69 to 165%) from the agreement (100%), which was followed by P (113 to 306%), whereas Fe showed the largest deviation (124 to 368%). CONCLUSIONS: Caution should be exercised in using FCTs for estimation of dietary Fe intake. The wide variation was not due to the use of the FCT established for each area, because the inter-study site variation within one area was as large as the inter-area variation.

Adult↗

DNA-DNA hybridization assay for detection of Salmonella spp. in foods.

We have developed a DNA-DNA hybridization test for the presence of Salmonella spp. in foods. This test requires an initial pre-enrichment of food samples in nutrient broth but does not require selective enrichment. Samples of food cultures are collected on membrane filters and assayed by molecular hybridization to labeled probes. The probes consist of DNA sequences which are unique to the genus Salmonella and are widely distributed in the genus. A diverse panel of foods was assayed successfully by this methodology.

Bacteriological Techniques↗

Detection of Staphylococcal enterotoxin B via biomolecular interaction analysis mass spectrometry.

Detection of Staphylococcus enterotoxin B (SEB) by biomolecular interaction analysis mass spectrometry (BIA/MS) is presented in this work. The BIA/MS experiments were based on a surface plasmon resonance (SPR) MS immunoassay that detects affinity-captured SEB both via SPR and by means of exact and direct mass measurement by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Experiments were performed with standard samples and food samples to assess the BIA/MS limit of detection for SEB and to set the experimental parameters for proper quantitation. Single and double SPR referencing was performed to accurately estimate the amount of the bound toxin. Reproducible detection of 1 ng of SEB per ml, corresponding to affinity capture and MS analysis of approximately 500 amol of SEB, was readily achieved from both the standard and mushroom samples. A certain amount of SEB degradation was indicated by the signals in the mass spectra. The combination of MS with SPR-based methods of detection creates a unique approach capable of quantifying and qualitatively analyzing protein toxins from pathogenic organisms.

Antigens, Bacterial↗

Methods for detecting food-borne enteroviruses.

A method previously reported for detecting virus in a model system composed of cottage cheese contaminated with coxsackievirus type A9 has been adapted to detecting selected strains of enteroviruses in a variety of foods. Bentonite is omitted and serum is added for extracting virus from low-protein foods. Samples of foods, usually 25 g, must contain at least 3 to 4 plaque-forming units for a 50% probability of detecting virus. Sensitivity in detecting echovirus type 6 was lower than that for the other viruses used. After extraction from potato salad, poliovirus type 2 was completely reactivated if it had been neutralized with coproantibody, but it was only partially reactivated if neutralized with hyperimmune rabbit serum.

Aluminum Silicates↗

Does further clean-up reduce the matrix enhancement effect in gas chromatographic analysis of pesticide residues in food?

Sample extracts of apples, peas, green beans, oranges, raspberries, clementines, carrots, and wheat obtained using the Food and Drug Administration (acetone extraction) and Canadian Pest Management Regulatory Agency (acetonitrile extraction) multiresidue methods for pesticides were subjected to clean-up using different solid-phase extraction (SPE) cartridges in an attempt to reduce or eliminate the matrix enhancement effect. The matrix enhancement effect is related to the blocking of active sites on the injector liner by matrix components, thereby increasing signal in the presence of matrix versus standards in solvent in which the pesticides themselves interact with the active sites. Graphitized carbon black (GCB) was often used in combination with various anion-exchange SPE cartridges. The extracts were then spiked with organophosphorus insecticides. These process standards were then compared to standards in acetone of the same concentration using gas chromatography with flame photometric detection or ion trap mass spectrometric detection. Sample matrix enhancement varied from little to no effect for some pesticides (e.g. chlorpyrifos, malathion) to >200% in the case of certain susceptible pesticides. The GCB removed color components but showed little effect in reducing matrix enhancement by itself. The anion-exchange cartridges in combination with GCB or not, substantially reduced the matrix enhancement effect but did not eliminate it.

Acetone↗

Plasticizers in total diet samples, baby food and infant formulae.

The plasticizers di-n-butylphthalate (DBP), butylbenzylphthalate (BBP), di-2-(ethylhexyl)phthalate (DEHP) and di-2-(ethylhexyl)adipate (DEHA) were analysed in 29 total diet samples, in 11 samples of baby food and in 11 samples of infant formulae. In all of the total diet samples the presence of one or more of the plasticizers was demonstrated. Maximum and minimum mean concentrations in the total diet samples were: 0.09-0.19 mg DBP/kg, 0.017-0.019 mg BBP/kg, 0.11-0.18 mg DEHP/kg and 0.13-0.14 mg DEHA/kg. One or more of the phthalates was also found in about 50% of the samples of baby food as well as in infant formulae. The calculated mean maximum intakes of the individual compounds from the total diet samples were below 10% of the restrictions proposed by the EU Scientific Committee for Food (SCF), and the spread in individual intakes was considerable. DEHP was the plasticizer determined most frequently and contributed the highest fraction of its tolerable daily intake (TDI). Hence, the maximum calculated intake of DEHP from single samples of the foodstuffs analysed could be up to one-third of the TDI. The calculated mean intake of DEHA was about 1% of the TDI with a maximum value of 13% of the TDI. Violations of the restrictions proposed by the EU Scientific Committee for Food (SCF) in the form of TDI values or specific migration limits were not found in this investigation.

Adipates↗

Salmonella-TEK, a rapid screening method for Salmonella species in food.

A micro-enzyme-linked immunosorbent assay (micro-ELISA) using the Salmonella-TEK screen kit was tested for the detection of Salmonella spp. in pure cultures as well as in 30 artificially contaminated food samples and in 45 naturally contaminated food samples. Different raw, fleshy foods and processed foods were used as test products. The artificially contaminated minced meat samples were preenriched in buffered peptone water, and after incubation, different selective enrichment broths were tested. The micro-ELISA optical density values after enrichment and isolation of the different broths were very analogous. The quickest method to detect Salmonella spp. in different foods is to enrich them with Salmosyst broth, which reduces the total analysis time to 31 h. The Salmonella-TEK kit for Salmonella spp. provides a promising test for the detection of Salmonella antigens in food even when they are present at a low concentration (1 to 5 CFU/25 g). The cross-reaction of the anti-Salmonella antibodies, especially to other gram-negative bacteria, is nil.

Enzyme-Linked Immunosorbent Assay↗

Monoclonal antibody-based dot-blot ELISA for the detection of Salmonella in foods.

Monoclonal antibody (MAb) produced to polysaccharides in the LPS molecule of salmonellae was used in a dot-blot ELISA for detecting Salmonella in 873 food samples, ie 100 fresh chicken, 261 frozen chicken, 78 pork, 84 beef, 100 hen eggs, 100 duck eggs, 50 sea-mussels, 50 shrimps and 50 squids in comparison with the conventional culture method. Salmonella culture from foods involved the following steps: pre-enrichment, enrichment in selective medium, isolation on selective and indicator media, followed by biochemical and serological identification of appropriate colonies, respectively. The whole culture procedure took 5 days. Food samples from the selective enrichment medium were also subjected to the MAb-based dot-blot ELISA. The whole procedure of dot-blot ELISA took less than 2 hours. Among 873 food samples, salmonellae could be recovered from 7.4% of the samples by the bacterial isolation method (16% of fresh chicken, 8.8% of frozen chicken, 24.4% of pork, 3.6% of beef and 2% each of hen eggs and duck eggs, respectively). Salmonella derby were predominant among pork samples while S.paratyphi B biovar java predominated in chicken. The MAb-based dot-blot ELISA were positive in 19.5% of the food samples, i.e. 30% of fresh chicken, 27.6% of frozen chicken, 34.6% of pork, 21.4% of beef, 20% of shrimp, 16% of sea-mussels, 2% of hen eggs and 4% of duck eggs. The sensitivity and specificity of the MAb-based dot-blot ELISA compared to the bacterial culture method were 81.5% and 85%, respectively. The discrepancy of the data between the culture method and the dot-blot ELISA might be due to the fact that the culture method could detect only living cells at numbers that gave at least one isolated colony on the selective/differential plate while the dot-blot ELISA detects any form of Salmonella antigen. The monoclonal antibody-based dot-blot ELISA offers several advantages over the conventional bacterial culture method when it is used for screening of Salmonella contamination in foods, especially export foods. These include rapidity, cost-effectiveness and simplicity (the dot-blot ELISA does not need highly trained personnel or equipment, in contrast to the culture method). The test can be performed in field conditions and the result can be read visually. It also offers multisample analysis at one time which renders more samples of food for screening possible, thus false negative results are fewer which, in turn, assures the quality of the export food in a cost-saving, short time frame.

Antibodies, Monoclonal↗

Validated PCR assay for the routine detection of Salmonella in food.

We developed a rapid and reliable PCR assay with genus-specific primers for the detection of Salmonella in food samples. With these primers, no primer-specific amplicons were detected when challenged with cultures of microorganisms other than salmonellae, and positive results, i.e., Salmonella-specific bands, were obtained with pure cultures of all 125 Salmonella isolates tested, which represented 100 serovars. The PCR assay was optimized using both pure cultures and artificially inoculated food samples. The assay results were compared with those of the Australian standard culture methods, using more than 500 "naturally" contaminated food samples, over a period of 9 years. Food samples were subjected to nonselective preenrichment in buffered peptone water followed by selective enrichment in Rappaport Vassiliadis (RV) broth and mannitol selenite cystine (MSC) broth. A simple sample preparation method was developed based on concentrating bacterial cells from 1 ml of RV or MSC broths. The PCR results were in perfect agreement with the results of the standard culture methods; no false-positive or false-negative results were obtained. However, the PCR assay was extremely rapid, and results could be obtained within 4 h of testing of enrichment broths.

Australia↗

Effects of wheat maturation stage and cooking method on dietary fiber and phytic acid contents of firik, a wheat-based local food.

Samples of two durum wheat cultivars (cvs. Duraking and Ege 88) at different maturation stages (13, 16, 19, 22, 25 days post anthesis) were processed into firik (a wheat-based specialty food) using two different cooking methods: roasting (scorching) on flames and boiling at atmospheric pressure. Both the acid detergent and neutral detergent fiber contents of the firiks produced from two durum wheat samples decreased significantly (p < 0.01) with maturation. Total P contents of the firiks of both cultivars produced by both methods showed a significant downward trend within the period of maturation while their phytic acid contents showed a significant upward trend (p < 0.01). It was possible to obtain a reduced phytic acid, high fiber product from the wheats harvested at early stages of maturation (13 and 16 days after anthesis).

Cooking↗

Use of polymerase chain reaction for detection of Listeria monocytogenes in food.

A previously described polymerase chain reaction (PCR) assay (B. Furrer, U. Candrian, C. Höfelein, and J. Lüthy, J. Appl. Bacteriol. 70:372-379, 1991) was used to analyze food for the presence of Listeria monocytogenes. Food samples were artificially contaminated to develop two procedures to detect the organism following enrichment steps. Procedure A was based on dilution of the enrichment broth followed by lysis of the bacteria and direct analysis of the lysate with PCR. With procedure A and artificially contaminated food samples, it was possible to detect fewer than 10 bacteria per 10 g of food. In procedure B, centrifugation was used to concentrate bacteria before lysis and PCR. With procedure A, 330 naturally contaminated food samples of several types were analyzed. Twenty samples were found to be positive for L. monocytogenes, which was in agreement with the classical culture technique. By using procedure B on a subset of 100 food samples, 14 were found to be positive by PCR whereas the classical culture method detected only 13. Analysis times, including enrichment steps, were 56 and 32 h with procedures A and B, respectively.

Animals↗

Sources of mutagenicity in cooked Finnish foods.

The mutagenic activities of the alkaline fractions derived from various heat-processed, ready-made meat, fish and poultry foods were studied using the Salmonella mutagenicity assay with strain TA98 and S-9 mix to provide information about the mutagenicity of everyday Finnish foods. The majority of the food samples tested were mutagenic. The mutagenic activities of various commercial ready-made products. Mutagenicity varied remarkably between different samples. The cooking temperature clearly affected the mutagenicities of fried or reheated food samples, mutagenicity increasing with increasing temperature. The results indicate that the main sources of cooked-food mutagens in everyday Finnish foods are grilled and broiled products and foods fried at restaurants and at home. By comparison, commercial ready-made fried foods are only a minor source of mutagenicity. Variations between equivalent food samples indicated that heat processing has a marked effect on the mutagenic activity of the product, which might therefore be reduced by modifying the cooking process.

Animals↗

Polybrominated diphenyl ethers contamination of United States food.

Elevated levels of polybrominated diphenyl ethers (PBDEs), a type of brominated flame retardant, were recently detected in U.S. nursing mothers' milk. These halogenated compounds chemically and toxicologically resemble others such as polychlorinated biphenyls (PCBs), whose route of intake is almost exclusively through food of animal origin. This study is the first to report the levels of PBDEs in U.S. foods in a market basket survey of 30 food types (total of 32 food samples) from three major supermarket chains in Dallas, TX. Food samples were almost exclusively foods of animal origin: meat, fish, and dairy products. Thirteen PBDE congeners were measured for each sample. Levels were then compared to existing PBDE food studies from other countries where available. In this study, levels of PBDEs are highest in fish, then meat, and lowest in dairy products; median levels were 1725 (range 8.5-3078), 283 (range 0.9-679), and 31.5 (0.2-1373), parts per trillion (ppt), or pg/g, wet weight, respectively. Nonfat milk did not have any detectable PBDE levels. In fish, PBDE congener 47 (2,2',4,4'-tetraBDE) contributes up to 70% of the total PBDEs, followed by congeners 100 (2,2',4,4',6) and 99 (2,2',4,4',5). In meat congener 99 predominates, followed by 47. In dairy, BDE 47 predominates followed by 99. U.S. food PBDE levels measured in this study are higher than reported in two other published market based studies from Spain and Japan. Although these findings are preliminary and will be updated with analyses of new samples, they suggest that food is a major route of intake for PBDEs.

Dairy Products↗

Enzyme-linked immunosorbent assay for rapid detection of toxins from Clostridium perfringens.

An enzyme-linked immunosorbent assay (ELISA) with antibodies specific to beta, epsilon and iota ib toxins of Clostridium perfringens was developed to detect beta, epsilon and iota ib toxins, respectively. The ELISA was sensitive enough to detect as little as 1.0 ng/ml of purified beta and iota ib toxins and 0.1 ng/ml of purified epsilon toxin. By means of the ELISA method, 192 isolates of C. perfringens from food samples in Japan and Thailand, and 58 isolates from patients suffering from gas gangrene or gastroenteritis were examined. One isolate from food samples in Japan, three from food samples in Thailand and five from stools of patients with gastroenteritis were C. perfringens type D. One type B and one type C were detected from the stools of patients with gastroenteritis.

ADP Ribose Transferases↗

Comparison of the mouse bioassay and enzyme-linked immunosorbent assay procedures for the detection of type A botulinal toxin in food.

Samples of chili linked to a foodborne illness outbreak of type A botulism were examined for preformed type A botulinal toxin using two enzyme-linked immunosorbent assay (ELISA) procedures and the mouse bioassay. One of the samples was positive for type A botulinal toxin and three of the samples were negative for type A, B, E, and F botulinal toxins using the three methods. The mouse bioassay indicated that type A toxin was present at the 10,000 minimal lethal dose per gram (MLD per g) of product. The ELISA tests indicated a toxicity of 7,650 MLD per g with one method and 8,350 MLD per g with the other method. The sample toxicity determined by the ELISA was estimated by comparing samples to a standard curve generated with standard type A neurotoxin in casein buffer. The ELISA methods are more rapid than the mouse bioassay, since the toxin type can be determined in 1 day. The mouse bioassay is more sensitive than the ELISA but usually requires multiple assays to obtain the toxin type and toxicity. Type A culture isolates from the sample were also verified using one ELISA method.

Animals↗

Screening of heat-processed Finnish foods for the mutagens 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine.

The concentrations of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and DiMeIQx (i.e. the sum of 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline and 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline) were determined as their 3,5-bis(trifluoromethyl)benzyl bromide derivatives in commercial heat-processed daily Finnish foods using the gas chromatography-mass spectrometry technique with negative chemical ionization. With this technique it was possible to detect picogram quantities of halogenated aminoimidazoazaarene (AIA) derivatives. The mutagenic activities of the food samples were tested using the Ames Salmonella assay. The mutagenicities and amounts of the AIA compounds analysed varied considerably in the food samples. All the flame-broiled fish samples and the majority of the grilled pork and chicken samples were positive in the test; of all the food samples tested fish samples had the highest mutagenic activity. Industrially produced meat patties were non-mutagenic. The amounts of MeIQx, DiMeIQx and PhIP in grilled products that exhibited mutagenic activity varied in the range 0.04 to 0.4, 0.03 to 0.2 and 0.5 to 3.8 ng per g cooked food, respectively. Some of the highly mutagenic flame-broiled fish samples contained only PhIP, the amount of which varied from 0.5 to 5.5 ng per g fish. One of the samples also contained a small amount of MeIQx. None of the non-mutagenic meat patties contained detectable amounts of MeIQx, DiMeIQx or PhIP. In this screening study evaluation of the occurrence of AIA compounds was carried out in a relatively wide range of commercial heat-processed Finnish foods. Considerable differences between equivalent products from different manufacturers were found in many cases. This variation indicates that industrial processing of food has a marked effect on the mutagenic activity of the product.

Animals↗

Determination of calcium intake of help of atomic absorption spectrophotometry.

In a dietary survey, large numbers of a wide variety of food samples have to be analyzed. The atomic absorption spectroscopy (AAS), with its simplicity, accuracy and rapidity, suggests itself for determination of many elements in such a case. In this investigation, calcium determination by AAS is reviewed. Analytical parameters, interferences and their control are studied. Food samples are dry ashed and treated so as to eliminate silicates. Most serious interference comes from phosphates and is successfully controlled by addition of lanthanum. Calcium is determined in the range 2-10 ppm with a sensitivity of 0.1 ppm. Single food samples and samples representing food mixtures are analyzed, and calcium intake of different population groups is computed. In most cases, results compare favourably well with those obtained from food composition tables. Calcium intake in the Netherlands both of the average per caput and of chosen population groups is found to cover the recommendations except for infants. Differences do exist between intakes of different groups and in different seasons.

Age Factors↗