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PubMed2004

Specific polyclonal-based immunoassays for sulfathiazole.

A highly sensitive and specific enzyme-linked immunosorbent assay has been developed for detection of sulfathiazole (STZ, 4-amino- N-thiazol-2-yl-benzenesulfonamide). A set of haptens was synthesized in order to produce polyclonal antibodies against sulfonamides. Two ELISA formats (antibody-coated and conjugate-coated) were also investigated using all the serum/coating conjugate combinations that showed specific recognition. The developed ELISA succeeded in detection of STZ at concentrations as low as 0.03 ng mL(-1) over a measurable range of 0.12-6.71 ng mL(-1). Selectivity studies have demonstrated that other sulfonamides do not interfere significantly (<10%) with analysis of STZ by this immunochemical technique. Analysis of spiked bee honey samples by the developed ELISA method showed recoveries were good. The selectivity and sensitivity (IC(50)=1.6 ng mL(-1)) make it a suitable screening method for determination of low levels of STZ in food samples.

Antibodies↗

PubMed1989

ELISA with enzyme amplification for sensitive detection of staphylococcal enterotoxins in food.

A highly sensitive amplified ELISA in microtitre plates for the detection of staphylococcal enterotoxins compared favourably with a commercially available kit. The amplified ELISA demonstrated a reduced effect of the food sample matrix and an ease of handling. Using the amplified ELISA, the influence of different parameters (pH, time, temperature, re-extraction, concentration procedure) on the recovery efficiency and assay reliability was tested on foods implicated in food poisoning outbreaks as well as on spiked foods containing low levels of enterotoxins.

Enterotoxins↗

PubMed1992

Principles involved in the detection and enumeration of clostridia in foods.

The clostridia are a group of anaerobic bacteria that vary considerably in their biochemical and physiological properties. Not surprisingly, attempts to develop a single isolation medium for all species that occur in foods have not been entirely successful, and the problem is compounded by the need to recover both vegetative cells and spores, some of the latter being unable to germinate without heat activation. Most available isolation media, except some of those used in the dairy industry, include sulphite and an appropriate iron salt, so that blackening due to sulphite reduction can serve as a differential test for clostridia. The limitations of this test in solid agar media are discussed and some advantages described in relation to its use in liquid media for Most Probable Number determinations. A medium favoured for the purpose is the Differential Reinforced Clostridial Medium of Gibbs and Freame (1965). An unresolved issue is whether or not special precautions are needed to exclude oxygen during food sample preparation and dilution, preparation of media, and in conditions used for anaerobic incubation. Although such stringency may be required for maximum recovery of sub-lethally damaged cells or spores, practical constraints in food control laboratories necessitate use of relatively simple procedures for detecting clostridia routinely.

Anaerobiosis↗

PubMed2002

Even small amounts of gluten cause relapse in children with celiac disease.

BACKGROUND: Previously, a gluten challenge was customary to establish the diagnosis of celiac disease in children. There are no clear recommendations on how to perform this challenge or what markers to rely on for timing the biopsy after the challenge. The authors' aim was to monitor gluten intake, clinical symptoms, and antibody kinetics to evaluate the influence of gluten exposure during the challenge. METHODS: Twenty-five children under investigation for suspected celiac disease were challenged. One child was excluded because blood samples, food records, or biopsy was lacking. Median age at the postchallenge biopsy was 3.8 (2.7-8.8) years. The families kept daily records of the children's gluten intake and of symptoms that occurred. Blood samples were taken monthly for analysis of antigliadin and endomysium antibodies and total immunoglobulin A (IgA). A third biopsy was performed when clinical symptoms suggested a relapse. RESULTS: All 24 children showed deterioration of the mucosa or elevated antibodies during gluten challenge. Median duration of the challenge was 13 (5-51) weeks, and mean gluten intake was 1.7 (0.2-4.3) g/d and 0.1 (0.02-0.26) g/kg daily. CONCLUSIONS: Gluten intake during the challenge varied widely, and the parents were unable to give their children the recommended amount. Despite the small amounts given, all children showed signs of relapse at a clinical, laboratory, or histologic level. Much smaller amounts of gluten than previously suggested seem sufficient to cause relapse during gluten challenge in children.

Biopsy↗

PubMed1991

Immunochemical assessment of mycotoxins in 1989 grain foods: evidence for deoxynivalenol (vomitoxin) contamination.

To assess the potential for mycotoxin contamination of the human food supply following the 1988 U.S. drought, 92 grain food samples were purchased from retail outlets in the summer of 1989 and surveyed for aflatoxin B1, zearalenone, and deoxynivalenol (DON [vomitoxin]) by monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Only one sample (buckwheat flour) was found to contain aflatoxin B1 (12 ng/g), whereas zearalenone was found in 26% of the samples at a mean concentration of 19 ng/g. In contrast, the DON ELISA was positive in 50% of the samples at a detection level of 1.0 micrograms/g. Between 63 and 88% of corn cereals, wheat flour/muffin mixes, rice cereals, and corn meal/muffin mixes yielded positive results for DON, whereas 25 to 50% of oat cereals, wheat- and oat-based cookies/crackers, corn chips, popcorn, and mixed-grain cereals were positive for DON. The mean DON content of the positive samples was 4.0 micrograms/g, and the minimum and maximum levels were 1.2 and 19 micrograms/g, respectively. When positive ELISA samples were also analyzed by high-performance liquid chromatography, a strong correlation between the two methods was found. The presence of DON in the two highest samples, corn meal and mixed-grain cereal, which contained 19 and 16 micrograms/g, respectively, was quantitatively confirmed by gas chromatography-mass spectrometry. The results indicated that DON was present in 1989 retail food products at concentrations that exceeded those found in previous market surveys and that have been experimentally associated with impaired animal health.

Aflatoxin B1↗

PubMed2003

Genetic diversity of Campylobacter jejuni isolates from farm animals and the farm environment.

The genetic diversity of Campylobacter jejuni isolates from farm animals and their environment was investigated by multilocus sequence typing (MLST). A total of 30 genotypes, defined by allelic profiles (assigned to sequence types [STs]), were found in 112 C. jejuni isolates originating in poultry, cattle, sheep, starlings, and slurry. All but two of these genotypes belonged to one of nine C. jejuni clonal complexes previously identified in isolates from human disease and retail food samples and one clonal complex previously associated with an environmental source. There was some evidence for the association of certain clonal complexes with particular farm animals: isolates belonging to the ST-45 complex predominated among poultry isolates but were absent among sheep isolates, while isolates belonging to the ST-61 and ST-42 complexes were predominant among sheep isolates but were absent from the poultry isolates. In contrast, ST-21 complex isolates were distributed among the different isolation sources. Comparison with MLST data from 91 human disease isolates showed small but significant genetic differentiation between the farm and human isolates; however, representatives of six clonal complexes were found in both samples. These data demonstrate that MLST and the clonal complex model can be used to identify and compare the genotypes of C. jejuni isolates from farm animals and the environment with those from retail food and human disease.

Alleles↗

PubMed2006

An overview on GSF activities at the Semipalatinsk Test Site, Kazakhstan.

The Semipalatinsk Test Site (STS) in Kazakhstan was one of the major sites used by the former USSR for testing nuclear weapons for more than 40 years. Since the early 1990s, agricultural activities have been re-established there by neighbouring collective and private farms. Therefore, it has become important to evaluate the radiological situation and the current and future risk to people living on and using the contaminated area. During the last eight years, GSF has participated in many international projects performed on the STS to evaluate the radiological situation. A large number of soil, vegetation and food samples has been collected and analysed. Internal dose is one of the main components of the total dose when deriving risk factors for a population living within the test site. Internal doses, based on food monitoring and whole body measurements, were calculated for adults and were in the range of 13-500 microSv/y due to radiocaesium and radiostrontium.

Animals↗

PubMed2000

Prevalence of enterotoxigenic motile aeromonads in children, fish, milk and ice-cream and their public health significance.

Motile aeromonads, which have been implicated as causative agents of human gastroenteritis, meningitis. endocarditis and osteomyelitis, were isolated from fish, pasteurized and flavored milk and ice creams. Forty-five isolates of Aeromonas from diarrheic children (100) and 285 food samples comprising of fish (100), milk (85) and ice creams (100) were identified as A. hydrophila (21), A. sobria (16) and A. caviae (8). Five of these 45 isolates of Aeromonoas (3 A. sobria, 2 A. hydrophila) were from fecal samples of diarrheic children under five years of age. Twenty-six of these 45 isolates were found to be enterotoxin producing by ligated rabbit ileum loop technique. Isolation of enterotoxigenic motile aeromonads from food as well as diarrheic children have great public health significance which will be further discussed.

Aeromonas hydrophila↗

PubMed1998

Detection of hemolysin variants of Shiga toxin-producing Escherichia coli by PCR and culture on vancomycin-cefixime-cefsulodin blood agar.

The presence of a hemolysin-encoding gene, elyA or hlyA, from Shiga toxin-producing Escherichia coli (STEC) was detected by PCR in each of 95 strains tested. PCR products of elyA from human STEC isolates of serovars frequently detected in Germany, such as O157:H-, O103:H2, O103:H-, O26:H11, and O26:H-, showed nucleotide sequences identical to previously reported ones for O157:H7 and O111:H- strains. Compared to them, four elyA amplicons derived from human isolates of rare STEC serovars showed identity of about 98% but lacked an AluI restriction site. However, the nucleotide sequence of an amplicon derived from a porcine O138:K81:H- STEC strain was identical to the corresponding region of hlyA, encoding alpha-hemolysin, from E. coli. This hlyA amplicon showed 68% identity with the nucleotide sequence of the corresponding elyA fragment. It differed from the elyA PCR product in restriction fragments generated by AluI, EcoRI, and MluI. Of the 95 representative STEC strains, 88 produced hemolysin on blood agar supplemented with vancomycin (30 mg/liter), cefixime (20 micrograms/liter), and cefsulodin (3 mg/liter) (BVCC). The lowest added numbers of two to six STEC CFU per g of stool or per ml of raw milk were detectable on BVCC plates after seeding of the preenrichment broth, modified tryptic soy broth (mTSB) supplemented with novobiocin (10 mg/liter), with 16 STEC strains. These strains represented the seven prevailing serovars diagnosed from German patients. However, with ground-beef samples, PCR was essential to identify the lowest added numbers of two to six STEC CFU among colonies of hemolyzing Enterobacteriaceae, such as Serratia spp. and alpha-hemolysin-producing E. coli. We conclude that preenrichment of stool and food samples in mTSB for 6 h followed by overnight culturing on BVCC is a simple method for the isolation and presumptive identification of STEC.

Bacterial Toxins↗

PubMed1993

Import control of irradiated foods by the thermoluminescence method.

A thermoluminescence (TL) method was applied for the import control of irradiated foods. The method is based on the determination of the TL of mineral contaminants in foods. Detection of irradiation was incorporated in official Finnish control procedures in spring 1990. For foodstuffs with a reduced microbe content and in which no fumigant residues are found, possible irradiation is investigated by the TL method. The minerals are separated from the foods in different ways: picking is used for spices; water rinsing for herbs, spices, berries and mushrooms; high-density liquid to separate the organic material from the mineral fraction in seafood; and carbon tetrachloride for foods forming gels with water. To date about 140 food samples have been analysed for control purposes: 50 samples of herbs and spices, 25 samples of berries and mushrooms and 65 samples of seafood. Of these, 14 samples of herbs and spices and 5 samples of seafood were shown to have been irradiated. Differences in TL intensity between irradiated and unirradiated samples were at least 1 and usually 3-4 orders of magnitude.

Animals↗

PubMed1986

Natural radiation environment of the Nasik and Neyveli thermal power stations, India.

The trace quantities of naturally occurring radionuclides present in the coal fuel of thermal power stations (TPS) result in redistribution of these radionuclides in the vicinity of the power stations through the deposition and disposal of ash. Two large size TPSs have been studied for environmental radioactivity and radiation doses in the vicinity of the stations. The stations were selected on the basis of the levels of primordial radionuclides in the fuel used. A minor increase was observed in the radiation levels in the ash-pond areas of the TPSs. A few food samples grown in TPS surroundings showed marginal increases in their natural radioactivity content.

Coal↗

PubMed2000

Gas chromatographic quantification of amino acid enantiomers in food matrices by their N(O,S)-ethoxycarbonyl heptafluorobutyl ester derivatives.

Several amino acid enantiomer derivatives were prepared with different chloroformates and analysed by gas chromatography (GC) on a Chirasil-L-Val GC column, at a temperature below 200 degrees C. Among them the N(O,S)-ethoxycarbonyl heptafluorobutyl esters presented the best compromise between short retention times, high yield responses and good resolution for almost all the tested amino acids. These derivatives proved to be suited for quantification of amino acids in aqueous media, with L-p-chlorophenylalanine as internal standard. The developed procedure was applied to several food samples for determination of their free amino acid profiles.

Amino Acids↗

PubMed2004

Generic lanthanide fluoroimmunoassay for the simultaneous screening of 18 sulfonamides using an engineered antibody.

Sulfa antibiotics (sulfonamides) are used in veterinary and human medicine for therapeutic and prophylactic purposes. Veterinary use can result in foodstuffs derived from animals being contaminated with residual sulfonamides. Current sulfonamide-screening methods (mainly based on bacterial growth inhibition) are slow and inaccurate, since sensitivities of bacteria to different sulfonamides vary a lot. Therefore, a rapid immunoassay that was able to detect at least 18 different sulfonamides at the MRL level (100 microg/kg) from food samples in a single reaction was developed. The assay was reproducible and adequately accurate for screening purposes. The presence of sulfonamide metabolites did not cause major assay interference. We also demonstrated reliable detection of sulfonamides from a panel of meat, milk, and serum samples with the assay.

Animals↗

PubMed1998

Survey of fungal counts and natural occurrence of aflatoxins in Malaysian starch-based foods.

In a survey of starch-based foods sampled from retail outlets in Malaysia, fungal colonies were mostly detected in wheat flour (100%), followed by rice flour (74%), glutinous rice grains (72%), ordinary rice grains (60%), glutinous rice flour (48%) and corn flour (26%). All positive samples of ordinary rice and glutinous rice grains had total fungal counts below 10(3) cfu/g sample, while among the positive rice flour, glutinous rice flour and corn flour samples, the highest total fungal count was more than 10(3) but less than 10(4) cfu/g sample respectively. However, in wheat flour samples total fungal count ranged from 10(2) cfu/g sample to slightly more than 10(4) cfu/g sample. Aflatoxigenic colonies were mostly detected in wheat flour (20%), followed by ordinary rice grains (4%), glutinous rice grains (4%) and glutinous rice flour (2%). No aflatoxigenic colonies were isolated from rice flour and corn flour samples. Screening of aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2 using reversed-phase HPLC were carried out on 84 samples of ordinary rice grains and 83 samples of wheat flour. Two point four percent (2.4%) of ordinary rice grains were positive for aflatoxin G1 and 3.6% were positive for aflatoxin G2. All the positive samples were collected from private homes at concentrations ranging from 3.69-77.50 micrograms/kg. One point two percent (1.2%) of wheat flour samples were positive for aflatoxin B1 at a concentration of 25.62 micrograms/kg, 4.8% were positive for aflatoxin B2 at concentrations ranging from 11.25-252.50 micrograms/kg, 3.6% were positive for aflatoxin G1 at concentrations ranging from 25.00-289.38 micrograms/kg and 13.25% were positive for aflatoxin G2 at concentrations ranging from 16.25-436.25 micrograms/kg. Similarly, positive wheat flour samples were mostly collected from private homes.

Aflatoxins↗

PubMed1978

New medium for rapid screening and enumeration of Clostridium perfringens in foods.

A rapid and sensitive procedure for estimating low numbers of Clostridium perfringens has been investigated and compared to methods used currently in the food industry. The new liquid medium, RPM (rapid perfringens medium), was compared with sulfite-polymyxin-sulfadiazine agar and tryptose-sulfite-cycloserine agar in recovery studies with naturally contaminated and with inoculated foods. The medium consists of a mixture of litmus milk and fluid thioglycolate medium fortified with glucose, peptone, gelatin, yeast extract, sodium chloride, and ferrous sulfate. Selectivity is based on an antibiotic system (polymyxin B sulfate and neomycin sulfate) incorporated into the medium, coupled with an incubation temprature of 46 to 48 degrees C for 24 h. Tubes were scored as positive if a stormy fermentation was observed. All tubes demonstrating stormy fermentation were confirmed as containing C. perfringens. Of a total of 774 naturally contaminated food samples, 546 samples (71%) were found to contain C. perfringens with RPM, whereas only 168 (22%) of the samples were positive using sulfite-polymyxin-sulfadiazine agar. C. perfringens was isolated from 71% of 85 other samples using RPM as compared to 14% with tryptose-sulfite-cycloserine agar. Enumeration studies on 14 individual samples using the most probable number technique also demonstrated greater sensitivity with RPM.

Clostridium perfringens↗

PubMed1997

Construction of specific DNA probe for the detection of Salmonella in food.

The Salmonella specific DNA fragment from genomic DNA of S. typhimurium ATCC 23566 was cloned in E. coli and successfully used as a digoxigenin labeled probe for detecting the presence of Salmonella serotypes in both artificially contaminated food and natural contaminated food samples.

Cloning, Molecular↗

PubMed1983

Selenium status of a rural (predominantly Amish) community living in a low-selenium area.

Selenium intake and blood selenium and selenium-dependent glutathione peroxidase concentrations were assessed in a rural, 71% Amish, sample of residents in a region with low availability of soil selenium and in urban (Columbus, OH) residents. Subjects were interviewed (24-h dietary recalls) on three separate occasions over an 18-month period, and blood and food samples were taken for analysis (maximum subject observations = 452). Mean selenium intake of the entire population was 82.8 +/- 4.4 micrograms/day. Rural and urban residents at 82.0 +/- 4.9 and 83.6 +/- 4.4 micrograms/day, respectively, did not differ significantly in intake. A breakdown by gender within location demonstrated that rural males consumed the most selenium (104.7 +/- 6.1 micrograms/day) and rural females the least (59.3 +/- 6.3 micrograms/day). Rural subject groups had significantly lower mean whole blood, plasma, and erythrocyte selenium levels (12.5 to 14.5%) as well as blood selenium-dependent glutathione peroxidase concentrations (5.6 to 10.8%) than urban groups. Subjects in both cohorts were in adequate selenium status as judged by blood parameters, even though about one-fourth of the dietary observations were below the safe and adequate range of selenium intake established by the Food and Nutrition Board. The rural diet was 18.3% lower in selenium density than the urban diet. High selenium grain products (greater than 0.2 micrograms/g) were the most important dietary selenium source in both groups, providing 29.3 to 41.6% of total intake.

Adolescent↗

PubMed2005

Campylobacter spp. in New Zealand raw sheep liver and human campylobacteriosis cases.

Sheep liver samples were tested for the presence and numbers of Campylobacter jejuni and C. coli during both spring and autumn. Over the same period, isolates were obtained from human clinical cases from the same geographical area as where the food samples were purchased. A subset of the C. jejuni isolates was typed by both Penner serotyping and pulsed field gel electrophoresis using the restriction enzyme SmaI, to estimate the proportion of liver isolate types that were also isolated from human cases of campylobacteriosis. Of the 272 liver samples tested, 180 (66.2%) contained Campylobacter. Most of the positive samples contained <3 MPN/g of the organism, and only 12 (6.7%) were contaminated at a level exceeding 100 MPN/g. A total of 180 C. jejuni isolates were obtained from sheep liver and another 200 from human faeces. Of these, 212 isolates were randomly selected for typing, half from raw liver and half from human faeces. More than half (61.1%) of the 106 C. jejuni isolates from liver were of subtypes that were also isolated from human cases. While the C. jejuni present in sheep liver were mostly of subtypes also isolated from human cases, the significance of this food as a vehicle of human campylobacteriosis needs to be examined further in respect to other factors such as dose-response information, consumption data, frequency of undercooking and cross contamination.

Animals↗