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Determination of starane (fluroxypyr) herbicide using flow injection spectrophotometry.

The starane herbicide was spectrophotometrically determined by the diazotization method in a flow injection assembly. Since starane is a substituted pyridyl compound the NH2 group at the p-position was exploited for diazotization. Starane was diazotized with nitrite and the diazotized product is coupled with beta-naphthol. The absorbance of the resulting azo dye was measured at 395 nm with a molar absorptivity of 1.5 x 10(4) L mol(-1) cm(-1). The calibration graph was linear over the range of 0.6 to 10 microg/mL, with a relative standard deviation (RSD) of 1.67% and a sampling through put of 60 samples h(-1). The % recovery for the determination of starane was found to be 96%. The method was successfully applied to the determination of the active ingredient of starane herbicide in its formulation as well as in food samples.

Acetates↗

A proposal for using previous experience in designing microbiological sampling plans based on variables.

A microbiological sampling system on the basis of variables is presented which requires only small numbers of replicates (n greater than or equal to 2). The system uses previous data on standard deviations of numbers of micro-organisms in batches and is particularly useful for in-plant situations. The discriminating power of the system is comparable to that of the current International Committee on Microbiological Specifications for Foods sampling schemes but uses more replicates.

Bacteriological Techniques↗

Determination of plasticisers in food by gas chromatography-mass spectrometry with ion-trap mass detection.

In this paper a method for the determination of plasticisers in food by gas chromatography-mass spectrometry with ion-trap mass detection is described. The plasticisers were quantified by an internal standard addition method using diisobutyl phthalate as the internal standard. Four selected food samples were spiked with nine different plasticisers at about 0.3 microgram/g. The recoveries of the plasticisers were in the range of 90 to 106%. The proposed method shows an improvement in precision and exhibits good linearity over a wide concentration range. The new method was applied to the analysis of real samples and the results were found to agree with those obtained using the well-established isotope dilution technique.

Food Analysis↗

Trace-level determination of pesticides in food using difficult matrix introduction-gas chromatography-time-of-flight mass spectrometry.

A procedure for the fully automated analysis of food samples by means of difficult matrix introduction-gas chromatography-time-of-flight mass spectrometry (DMI-GC-ToF MS) is discussed. After extraction, samples require very little clean-up and are injected in a micro- or micro-vial which is held in a liner. Next, the liner is placed in the injector and the contents of the vial are thermally desorbed and led directly to the capillary GC column. After GC-ToF MS analysis, the data are processed automatically using a peak deconvolution algorithm. The practicability of the procedure was demonstrated by analysing spiked grape and pineapple samples down to the 1-10 ng/g concentration level.

Food Analysis↗

Improved detection of Salmonella spp. in foods by fluorescent in situ hybridization with 23S rRNA probes: a comparison with conventional culture methods.

This report describes a new technique for the detection and identification of Salmonella species in food with the use of fluorescent in situ hybridization (FISH) with 23S rRNA-targeted oligonucleotide probes. Two species-specific 23S rRNA-targeted oligonucleotide probes (Sal-1 and Sal-3) were selected, and one (Sal-544) was newly designed. The relative specificities of these probes were compared with those of bacterial 23S rRNA sequences from the GenBank database and tested by in situ hybridization with bacterial cell smears of pure cultures. Fifty-one tested reference strains of Salmonella serovars belonging to subspecies I (enterica) hybridized with these probes. No cross-reactions with 46 other strains of the family Enterobacteriaceae or with another 14 bacterial strains from other families were observed. Storage of a Salmonella Panama test strain under various environmental conditions (2, 5, and 15% NaC1; -20 degrees C, 4 degrees C, and room temperature; pHs of 3.3 to 7.4) did not adversely affect the FISH method. No matrix effects were observed with 18 different kinds of foods. FISH was able to detect Salmonella spp. in 52 (probe Sal-1), 56 (probe Sal-3), and 35 (probe Sal-544) of 225 naturally contaminated food samples after 16 h of incubation in a preenrichment broth. When conventional culture and detection methods were used, Salmonella could be isolated from only 30 of these 225 samples. In contrast, FISH failed to identify Salmonella in only two of the culture-positive samples when Sal-1 and Sal-3 were used and in only three of the culture-positive samples when Sal-544 was used.

Animals↗

Design and application of a Staphylococcus-specific single strand conformation polymorphism-PCR analysis to monitor Staphylococcus populations diversity and dynamics during production of raw milk cheese.

AIM: Development of a nested-PCR single strand conformation polymorphism (SSCP) assay targeting the 16S rRNA genes of the Staphylococcus genus, to monitor staphylococci in cheese. METHODS AND RESULTS: New primer sets to specifically amplify 16S rDNA of staphylococci were designed to be used in a nested-PCR SSCP assay. The method was efficient in discriminating the staphylococcal species most frequently found in cheese. It was validated by monitoring Staphylococcus populations in three productions of raw milk cheese. Analysis of milk samples revealed dominant SSCP peaks corresponding to Staphylococcus aureus, Staphylococcus equorum and Staphylococcus saprophyticus. After 12 h, the S. aureus peak became dominant. CONCLUSIONS: The combination of specific Staphylococcus nested-PCR and SSCP allows rapid and direct monitoring of staphylococci diversity and dynamics in milk and cheese. In the core of the cheeses studied, S. aureus may have ecological advantages against other Staphylococcus populations. SIGNIFICANCE AND IMPACT OF THE STUDY: This approach is a promising tool to study the ecology of staphylococci in cheeses and in other food samples.

Animals↗

[Occurrence of aflatoxin-forming molds in foods].

Studies on the mould infection of nearly 900 food samples, performed by means of the screening test according to de Vogel et al. [2], showed that the occurrence of aflatoxin-forming moulds must be considered mainly with peanuts and other seeds, but also with any other food. From this point of view, all measures against mould growth in foods are of special importance.

Aflatoxins↗

Lipid content and fatty acid composition in foods commonly consumed by nursing Congolese women: incidences on their essential fatty acid intakes and breast milk fatty acids.

The fat content and fatty acid (FA) composition of nearly 40 foods, currently consumed by 102 nursing Congolese mothers living in Brazzaville, were determined to assess their impact on mothers' essential fatty acid (EFA) intakes and breast milk FA. Data on mothers' milk FA and dietary habits which allowed food selection were recently published (Rocquelin et al., 1998). Most foods were locally produced. Food samples were collected at local markets, bleached if necessary to avoid microbial degradation, and stored at +4 degrees C or -20 degrees C. They were lyophilized upon their arrival in the laboratory before lipid analyses. FA composition of food lipids was determined by capillary gas chromatography. Staple diets included low-fat, high-carbohydrate foods (processed cassava roots, wheat bread) and high-polyunsaturated fatty acid (PUFA) foods: soybean oil (high in 18 : 2 n-6 and alpha-18 : 3 n-3), bushbutter (dacryodes edulis), peanuts, avocado (high in fat and 18 : 2 n-6), freshwater and salt-water fish (high in LC n-3 and/or n-6 PUFA), and leafy green vegetables (low in fat but very high in alpha-18 : 3 n-3). Their frequent consumption by nursing mothers provided enough EFA to meet requirements due to lactation. It also explains why mothers' breast milk was rich in C8-C14 saturated FA (26% of total FA) and in n-6, n-3 PUFA (respectively 15.0% and 2.4% of total FA) highly profitable for breastfed infants' development. From this point of view, dietary habits of Congolese mothers have to be sustained for they are more adequate than most Western-type diets.

Animals↗

Sensitivity, specificity, and predictive values of three Salmonella rapid detection kits using fresh and frozen poultry environmental samples versus those of standard plating.

To reduce human exposure to Salmonella spp. in poultry products, broiler chicken flocks have been tested by culture methods. Since the standard techniques may take 3 to 5 days, rapid detection methods have been developed. In this study we tested the performance of three rapid tests originally developed for food samples by using environmental samples obtained from poultry houses. These rapid tests were Reveal, an enzyme-linked immunosorbent assay from Neogen Corp.; BIND, a bacterial ice nucleation detection method from Idetek Corp.; and a filter monitor method from Future Medical Technologies, Inc. For the standard culture, brilliant green with novabiocin and xylose-lysine-tergitol-4 agar were used for presumptive identification, and identities were confirmed by using poly-O antisera. Environmental samples were collected from farms belonging to an integrated poultry company prior to chick placement and 1 week before slaughter. Sensitivities, specificities, and predictive values with 95% confidence intervals were calculated. Statistical differences were determined by using McNemar's chi square test. The sensitivities of the different tests were not stable, varying widely between sample times, and were affected by freezing of the samples. All of the rapid tests had low sensitivities, which led to many false-negative results. All tests were able to detect Salmonella spp. at a concentration of 10 CFU/ml in at least one of four trials. The BIND and Reveal tests were simple to use with multiple samples and reduced laboratory time by up to 1 day. Based on our results, we do not recommend that any of these rapid tests, in their present state of development, be utilized with environmental samples collected with drag swabs.

Animals↗

Development and validation of a quantitative method for determination of carmine (E120) in foodstuffs by liquid chromatography: NMKL Collaborative Study.

A liquid chromatographic method for quantitative determination of carmine (E120) in different foodstuffs is described. Qualitative and semiquantitative methods for analysis of carmine and other related dyes are well established. However, quantitative methods available are based mainly on enzymatic reactions that are time-consuming or specific for analysis of carminic acid in yoghurt. In the method developed and validated here, carminic acid is extracted by boiling the sample with HCl, purified on a solid-phase extraction cartridge, and injected on a C18 analytical column. The method was evaluated by an internal analytical quality control and a collaborative study in which 11 laboratories from the Nordic countries participated. The food samples analyzed were fruit jelly, liqueur, juice, yoghurt, and ice cream. Materials were distributed to participants as uniform level and split level. Validation showed that the proposed method is well suited for quantitative determination of carmine. The detection limit is 0.1 mg/L. The mean relative standard deviation for reproducibility varies from 7.9 to 11.7%. The proposed method is simple and relatively fast compared with previously published methods.

Animals↗

Enterotoxigenic Bacillus spp. DNA fingerprint revealed in naturally contaminated nonfat dry milk powder using rep-PCR.

Dry milk powders and functional ingredients frequently contain high levels of viable bacterial spores, some of which may result in growth of toxigenic Bacillus spp. in reconstituted and temperature-abused foods. Samples from nonfat dry milk (NFDM), infant milk formula (IMF), coffee creamer, lecithin, and cocoa powder were subjected to a short heat treatment followed by enrichment in tryptone phosphate glucose yeast extract (TPGY) broth at 32 degrees C for 12-25 hours to obtain cell densities of 10(6) CFU ml(-1). DNA was extracted using a modification of established protocol, leading to the development of an optimized method for each food system. Purified DNA was amplified by rep-PCR using extragenic sequence-targeting primers and optimized for each food. PCR fingerprints from each food were analyzed electrophoretically for banding patterns earlier correlated to that of enterotoxigenic Bacillus spp. and Bacillus cereus positive control DNA fingerprints. Reverse passive latex agglutination (RPLA) and Bacillus Diarrhoeal Enterotoxin Enzyme Linked Immunosorbent Assay (Tecra Diagnostics) confirmed the presence of HBL and NHE enterotoxin production in NFDM, Coffee creamer, infant milk formula, and two lecithin samples but not in cocoa powder. These results demonstrate the utility of rep-PCR not only as a tool for bacterial genotyping, but a unique means of quality control and hygiene monitoring in food microbiology.

Animals↗

Salmonella detection by the polymyxin-cloth enzyme immunoassay using polyclonal and monoclonal detector antibodies.

Several commercially available O-antigen polyclonal antisera and a monoclonal antibody to the core region of lipopolysaccharide (LPS) were examined as sources of detector antibodies in a polymyxin-cloth enzyme immunoassay (polymyxin-CEIA) for Salmonella. In this assay, polymyxin-coated polyester cloth captured the LPS antigens from Salmonella broth cultures, followed by immunoenzymatic detection of the captured LPS using specific antibodies. Pools of polyvalent antisera reacted with all of the Salmonella strains tested, but also gave cross-reactions with some non-Salmonella bacteria. On the other hand, the monoclonal antibody gave positive reactions with all of the Salmonella tested except serogroup O-strains, but did not react with any of the non-Salmonella bacteria. The monoclonal antibody supplemented with a single factor serogroup O:35 rabbit antiserum was able to detect the serogroup O-strains without causing any cross-reactions with the non-Salmonella bacteria. As an example of the applicability of this assay system, low levels of Salmonella cells spiked into various food samples were successfully detected after an overnight enrichment in broth.

Animals↗

The Gamma Contamination Food Factor.

We suggest that radioactive food contamination, as determined solely by a quantitative gamma-ray spectroscopic measurement, may, apart from the total activity per unit mass, be for quick reference conveniently characterized by another single figure which we call the "Gamma Contamination Food Factor" (GCFF). This factor may be defined as the ratio of the total specific activity of gamma-ray-emitting radionuclides in the food sample (except that of 40K) to the specific activity of 40K either in the sample itself or in an "average man". We discuss briefly the meaning and advantages of these definitions.

Food Contamination, Radioactive↗

Incidence of Clostridium perfringens in American foods.

Food samples were examined for the presence of Clostridium perfringens. A medium described by Mossel and later modified by Angelotti et al. was used for the detection and enumeration of C. perfringens. The incidence of C. perfringens observed in the foods examined was 6.1%. C. perfringens was recovered from 2.7% of the commercially prepared frozen foods, 3.8% of fruits and vegetables, 5.0% of spices, 1.8% of home-prepared foods, and 16.4% of raw meat, poultry, and fish.

Animals↗

[Determination of soybean isoflavones in foods by HPLC].

OBJECTIVE: To establish a method for the determination of soybean isoflavones including daidzein and genistein in foods by high performance liquid chromatography. METHODS: Soybean isoflavones in food samples were refluxed and extracted by 0.5 mol/L hydrochloric acid and ethanol. Soybean isoflavones were chromatographically separated on a C18 column with a mobile phase consisting of methanol and 0.01 mol/L ammonium acetate buffer solution (pH 4.5) (60 + 40), and then detected at 254 nm with ultraviolet-visible detector. RESULTS: The detection limits were 0.05 microg/ml for daidzein and 0.04 microg/ml for genistein, respectively. The spiked recoveries of the samples were 91.4%-113.1%. The within-day and between-day (n = 6) relative standard deviations were 1.1%-2.8% and 2.9%-5.1%, respectively. CONCLUSION: The method is simple, sensitive and applicable to the determination of soybean isoflavones in foods. It also provides an analytical method for isoflavones in functional food and the absorption of phytoextrogens from foods.

Chromatography, High Pressure Liquid↗

Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies.

Five monoclonal antibodies (MAbs) and chicken immunoglobulin (IgY) were developed by immunizing with flagella purified from Listeria monocytogenes 4b and the five MAbs have been confirmed to be specific against three different epitopes of flagellin. The antibodies showed specific reaction to Listeria genus and no cross-reactivity with other bacteria tested in this experiment including E.coli O157:H7 and Salmonella enteritidis. Sandwich enzyme-linked immunosorbent assays (ELISA) using the MAbs and IgY were developed to detect Listeria species and the sensitivity and specificity of the developed ELISA have been analyzed. The detection limit of ELISA using MAb 2B1 and HRP labeled IgY was 1 x 10(5) cells/0.1 ml at 22 degrees C. and 1 x 10(6) cells/0.1 ml at 30 degrees C. ELISA using the pair of MAbs (MAbs 2B1 and HRP labeled MAbs 7A3) detected up to 10(4) cells/0.1 ml at 22 degrees C and 30 degrees C. Detection limit of sandwich ELISA using IgY was 10 times lower than MAb pair. Using the developed ELISA, we could detect several Listeria contaminated in food samples after 48 h-culturing. In conclusion, both MAbs and IgY have been proved to be highly specific to detect Listeria flagella and the developed sandwich ELISA using these antibodies would be useful tool for screening Listeria spp. in food.

Animals↗

[Accumulated occurrence of illnesses with Salmonella enteritidis in hospitals and nursing homes in the district Oberallgaeu, Bavaria, in July 2004].

We report on the investigation of a Salmonella enteritidis outbreak in hospitals and nursing homes in the district Oberallgaeu, Bavaria, in July 2004. Affected by this outbreak were hospital patients, inhabitants of nursing homes, kitchen coworkers and maintenance personnel. Within this outbreak six deaths were observed. A forensic medical investigation of four deaths revealed two cases of Salmonella enteritis as the primary cause of death, another cause of death was due to other causes and one cause of death remained unclear. The microbiological investigation of stool samples linked all positive samples to an outbreak during this period. The epidemiological outbreak curve pointed towards a foodborne transmission of the pathogen. The infection period could be limited to the weekend of the 3./4.7.2004. All affected institutions were supplied by a catering service with several sites. All tested food samples were negative for Salmonella enteritis. Epidemiological investigations revealed that a contaminated pudding was probably responsible for the outbreak.

Adult↗

Application of solid-phase microextraction for determining phenylurea herbicides and their homologous anilines from vegetables.

Residues of metobromuron, monolinuron and linuron herbicides and their aniline homologous were analyzed in carrots, onions and potatoes by solid-phase microextraction (SPME) performed with a polyacrylate fiber. A juice was obtained from food samples that were further diluted, and an aliquot was extracted after sodium chloride (14%) addition and pH control. At pH 4 only the phenylureas were extracted. A new extraction at pH 11 allowed the extraction of phenylureas plus homologous aniline metabolites. Determination was carried out by gas chromatography with nitrogen-phosporus detection (NPD) the identity of the determined compounds was studied by gas chromatography-mass spectrometry. Limits of quantification (LOQs) obtained with NPD and MS (selected-ion monitoring) were in the microg/kg order allowing determination of maximum residue levels (MRLs) established in the Spanish regulations. MRLs ranged from 0.02 to 0.1 mg/kg depending on the kind of food and herbicide. Under the proposed conditions matrix effects were low enough to permit calibration with samples proceeding from ecological (non-pesticide treated) crops. Twelve commercial samples of each carrots, onions and potatoes were analyzed and only three samples of potatoes contained residues of linuron at levels below MRLs.

Aniline Compounds↗