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A comparison of sampling methods for airborne fungal spores during an outbreak of aspergillosis in the forest aviary of the North Carolina Zoological Park.

An outbreak of aspergillosis with the death of six birds in the North Carolina Zoological Park R. J. Reynolds Forest Aviary in the spring of 1993 led to an investigation of the concentration of Aspergillus fumigatus spores in the air. No Aspergillus sp. was found in the facility through use of the drop plate method (gravitometric sampling) along with swab-sampling of selected surfaces within the exhibit and plating of food samples and nesting material onto petri dishes of nutrient media. A number factors that could stress the avian population were identified. These included excessive heat in the upper portion of the aviary due to the failure of an air handling system, a malfunctioning cooling tower, and large numbers of visitors to the facility (an average of 3,500/day). In addition, the outbreak occurred during a period of increased nesting behavior. Sampling of the fungal population of the air was conducted 1 year later, when no disease was noted, to compare the sensitivity of the commonly used drop plate method (open plates of nutrient media) with a volumetric impaction method (Andersen N-6 Air Sampler). The volumetric method delivered quantitative as well as qualitative data and exhibited more sensitivity for fungal spores of size similar to those of Aspergillus sp.

Air Microbiology↗

Contamination of food and agro products in the Republic of Macedonia.

The Republic of Macedonia has 662,000 ha of cultivated land with well-organised agricultural production. It is a well known producer of agricultural products. From 1991 to 1999, authorized laboratories conducted systematic control of about 6,500 food samples in 1991 to 36,000 by 1999 which included a whole range of hygienic quality parameters including microbiological and chemical. Microbiological contamination of food was reported to have caused disease outbreaks in 1,578 people in 1995, which dropped to 126 people in 1999. The decreasing trend of samples unfit for human consumption was observed for all food groups. As for the chemical parameters, an increasing trend of contaminated samples was noted only for additives, with peaks reaching 6.9% in 1998 and 4.9% in 1999 of the total number of analysed samples.

Food Contamination↗

Canadian Total Diet Study in 1998: pesticide levels in foods from Whitehorse, Yukon, Canada, and corresponding dietary intake estimates.

The Canadian Total Diet Study is a national survey to determine the level of chemical contaminants in the Canadian food supply. Food samples were collected from Whitehorse, Yukon, supermarkets as part of the study in 1998. Whitehorse was chosen as a sampling centre, despite its small population (n = 19,000), to determine if residue levels were different in foods available in northern communities relative to levels observed in previous studies in the more populated south. Foods were prepared as for consumption before pesticide residue analysis. Residue levels observed in most foods were similar to levels observed in samples from previous surveys from southern Canadian cities. Malathion and DDE (1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene), a transformation product of DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl(ethane), were the two most frequently detected compounds (26.4 and 25.8%, respectively). The majority of pesticides, however, had a detection frequency of < 5%. In general, pesticides in food composites were well below maximum residue limits established in the Canadian Food and Drug Regulations. Chlorpropham and captan had the highest dietary intakes (2.16 and 1.94 micrograms (kg body weight-day)-1, respectively), based on the results from Whitehorse. No dietary intakes above the acceptable daily intakes, however, were observed for any of the 39 pesticides investigated in any age-sex category, where an acceptable daily intake has been proposed.

Adolescent↗

Development of chemiluminescent ELISAs to DDT and its metabolites in food and environmental samples.

Two enzyme-linked immunosorbent assays (ELISA) with chemiluminescent (CL) detection for the insecticide DDT and the group of DDT-related compounds have been optimized and characterized. Both conjugate-coated ELISAs are based on monoclonal antibodies (MAbs) of different specificity and homologous protein conjugates. Effects of several physicochemical factors (ionic strength, pH, Tween-20 and Bovine serum albumin (BSA) concentrations) and solvents (methanol, ethanol, acetone and N,N'-dimethylformamide) on the performance of the assays were studied and optimized. For the DDT-selective assay, the sensitivity, estimated as the I(50) value, was 0.6 microg/l, with a linear working range between 0.1 and 2 microg/l and a limit of detection of 0.06 microg/l. For the DDT group-selective assay, the sensitivity was 0.2 microg/l, with a linear working range between 0.07 and 1 microg/l and a limit of detection of 0.04 microg/l. CL-ELISAs were four times more sensitive compared to colorimetric ELISAs. Finally, both immunoassays were applied to the detection of DDT and group of DDT-related compounds in spiked real water, soil and food samples.

Cross Reactions↗

Improved method for simultaneous determination of ascorbic acid and dehydroascorbic acid, isoascorbic acid and dehydroisoascorbic acid in food and biological samples.

The total vitamin C amount in different food and plasma samples was determined by a dual detection system, after HPLC separation, with direct detection of ascorbic acid and indirect fluorimetric detection of dehydroascorbic acid after a post-column O-phenyldiamine derivatisation. The two active forms of vitamin C and their D-isomers were separated within 10 min. The repeatability was determined by measurement of several fruits and vegetables and ranged from 0.3 to 1.9% (relative standard deviation) for vitamin C. The reproducibility, based on double determinations, ranged from 1.9 to 3.6% for vitamin C, depending on the matrix. The reproducibility, based on several determinations of reference materials, ranged from 2.4 to 3.7% for ascorbic acid and from 4.3 to 5.8% for dehydroascorbic acid, again depending on the matrix.

Ascorbic Acid↗

Dietary intake in the lower Mississippi delta region: results from the Foods of our Delta Study.

OBJECTIVE: To collect and evaluate food intake data from a culturally diverse population and compare with national survey data. DESIGN: The Foods Of Our Delta Study was a baseline, cross-sectional survey that utilized random-digit dialing methodology to identify the sample. Food intake was obtained from a 24-hour dietary recall administered by computer-assisted telephone interview using the multiple-pass method. SUBJECTS/SETTING: One thousand seven hundred fifty-one adults and 485 children in the Lower Mississippi Delta (Delta) of Louisiana, Arkansas, and Mississippi. STATISTICAL ANALYSES PERFORMED: Comparisons of subsets within the Delta were made using weighted t tests. Comparisons of the Delta with the overall US population from the US Department of Agriculture Continuing Survey of Food Intakes by Individuals and with the Dietary Reference Intakes were made using independent sample z tests of weighted estimates. RESULTS: Energy intake did not differ between the Delta and the US populations. Intakes of protein were lower, fat higher, and certain micronutrients lower in Delta adults than in US adults. Delta adults had a 20% lower intake of fruits and vegetables than the US adults and generally poorer adherence to recommendations of the Food Guide Pyramid. African American Delta adults generally consumed less-optimal diets than white Delta adults. Delta children had diets similar to children of the Continuing Survey of Food Intakes by Individuals sample population, but lower intakes were noted for vitamins A, C, riboflavin, and B-6, and for calcium and iron. APPLICATIONS/CONCLUSIONS: Data such as these will help drive intervention development in this rural region and perhaps set the stage for research in similarly impoverished areas.

Adolescent↗

Levels of polychlorinated biphenyls (PCBs) in meat and meat products from the state of Rio Grande do Sul, Brazil.

The levels of six polychlorinated biphenyl congeners (PCBs) were evaluated in 55 samples of meat (bovine and pork) and meat products (sausage, hot dog sausage, bologna sausage, canned export meat and salami) from 11 cities of the state of Rio Grande do Sul, Brazil, between July and August 2002. PCB congeners were found (in fat basis) in the following rank 52 (5.18 ng/g) > 180 (1.69 ng/g) > 10 (1.35 ng/g) > 28 (1.19 ng/g) >153 (0.47 ng/g) >138 (0.43 ng/g), with a summation SigmaPCB amounting to 10.30 ng/g. Meat products had higher PCB levels than meat. PCB levels in samples followed the rank: mixed meat products > pork meat > bovine meat. These results indicate the presence of PCBs in food samples from Rio Grande do Sul, Brazil, but the levels found were well below the maximum level established for animal food products in Brazil (3000 ng/g fat). Only one sample exceeded the maximum level established by the European Community (200 ng/g fat). This is the first paper describing background concentrations of PCBs in meat and meat products from Brazil.

Animals↗

Sensitive Listeria spp. immunoassay based on europium(III) nanoparticulate labels using time-resolved fluorescence.

Listeria spp. are Gram-positive rod shaped bacteria found universally in the environment. Pathogenic Listeria monocytogenes is seldom harmful to healthy adults, but can cause serious disease, listeriosis, especially to pregnant women, neonates, and elderly or immunocompromised people. Conventional methods for screening Listeria in food samples are time consuming and laborious, involving the use of a range of liquid media and plate cultures. In the current study, the total analysis time was shortened by employing a sensitive Listeria assay, which was able to detect the bacteria in low concentrations. Sensitivity of the sandwich immunoassay was substantially improved by utilizing europium(III)-chelate containing latex nanoparticles as tracers. Each 107 nm nanoparticle contained approximately 31000 europium(III)-chelates which enhanced the specific activity of the label. The sensitive nanoparticulate immunoassay developed for Listeria spp. was performed in one-step and two-step formats. One-step assay was notably faster, 15 min, and simpler to execute having analytical sensitivity of 300 CFU/ml and a dynamic range of three orders of magnitude. The sensitivity, 20 CFU/ml, of the 4 h two-step assay clearly exceeded that of the one-step assay, and the dynamic range was nearly five orders of magnitude. Food and environmental samples were measured against a commercial L. monocytogenes immunoassay with good correlation. The developed sensitive assay enabled shorter sample enrichment times and, therefore, faster analysis of Listeria spp. Obviously the detection of several other bacteria can also be enhanced by applying the nanoparticle assay technology.

Colony Count, Microbial↗

Determination of methyl bromide in foods by headspace capillary gas chromatography with electron capture detection.

Methyl bromide (MB, bromomethane) is determined in a variety of foods by headspace capillary gas chromatography with electron capture detection. The comminuted food sample as an aqueous sodium sulfate slurry is equilibrated with stirring for 1 h at room temperature before a 1 mL headspace aliquot is removed and injected using a modified on-column syringe needle. Methyl bromide is cryogenically focussed at -60 degrees C and then eluted by temperature programming. The procedure requires blending of soft samples, e.g. raisins, prunes, or oranges, and ultrasonic homogenization of hard samples, e.g. wheat, cocoa beans, corn, or nuts, with portions of water and ice so the final temperature of the food-water slurry is less than 1 degree C. A 20 g aliquot (4 g food) is then added to a cold headspace vial containing 4 g sodium sulfate. Losses of MB during a 3.5 min ultrasonic homogenization of wheat were 11% at 0.95 ppb and 4.4% at 4.8 ppb. For flour, cocoa, and finely divided spices, which do not require blending, 4 g is added to the cold headspace vial containing 16 mL cold water and 4 g sodium sulfate. Studies show that comminution of wheat or peanuts must be carried out to release MB trapped within the food so the headspace equilibrium can be attained in 1 h as well as to obtain homogeneous samples and representative sampling. No interferences were noted with the above foods or with many grain-based baking mixes analyzed.(ABSTRACT TRUNCATED AT 250 WORDS)

Cheese↗

Multiresidue analysis of pesticides in vegetables and fruits using two-layered column with graphitized carbon and water absorbent polymer.

A high-throughput multiresidue analysis of pesticides in non-fatty vegetables and fruits was developed. The method consisted of a single extraction and a single clean-up procedure. Food samples were extracted with ethyl acetate and the mixture of extract and food dregs were poured directly into the clean-up column. The clean-up column consisted of two layers of water-absorbent polymer (upper) and graphitized carbon (lower), which were packed in a reservoir (75 ml ) of a cartridge column. The polymer removed water in the extract while the carbon performed clean-up. In a recovery test, 110 pesticides were spiked and average recoveries were more than 95% from spinach and orange. Most pesticides were recovered in the range 70-115% with RSD usually < 10% for five experiments. The residue analyses were performed by the extraction of 12 pesticides from 13 samples. The two methods resulted in similar residue levels except chlorothalonil in celery, for which the result was lower with the proposed method. The results confirmed that the proposed method could be applied to monitoring of pesticide residue in foods.

Food Contamination↗

Dietary selenium intake by men and women in high and low selenium areas of Punjab.

OBJECTIVE: To determine the selenium intake of adults residing in high and low selenium areas of rural Punjab. DESIGN: All food samples consumed by the subjects were collected and analysed for selenium content. Based on food intake data and selenium content of foods, the selenium intake of the subjects was calculated. Hair, fingernails and urine samples from a sub-sample of subjects were collected and analysed for selenium. SETTING: Three villages from the selenium-endemic area of Nawan Shahr District and two villages from the non-endemic area of Ludhiana District, Punjab, India, were covered. SUBJECTS: Forty families from each of the two areas, with one adult male and one adult female in the age range of 20-40 years, were surveyed. Thus a total of 80 men and 80 women constituted the study sample. RESULTS: In the selenium-endemic area, the average selenium intake of both men and women was more than nine times that in the non-endemic area and exceeded the maximum tolerable limit in more than 60% of men. Mean selenium content of the hair, nails and urine of both men and women was tens of times higher than in the non-endemic area. CONCLUSIONS: High selenium intake in the endemic area resulted in high selenium content in the hair, nails and urine of men and women. In addition, clinical symptoms of selenium toxicity were also observed in some of the subjects. Selenium intake in the non-endemic area was marginally below the suggested value. Based on the study results, steps need to be taken to educate the public in the endemic area to avoid selenium toxicity.

Adult↗

Sample preparation methods for PCR detection of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on beef chuck shoulder using a single enrichment medium.

To improve the utility of the polymerase chain reaction (PCR) for food samples, methods for preparing template DNA were developed to remove PCR inhibitors. Beef chuck shoulder medallions, artificially contaminated, individually or in combination, with Escherichia coli serotype O157:H7 strain FSIS 45753-35, Salmonella typhimurium DT104 strain 13HP, or Listeria monocytogenes strain Scott A at concentrations of 10, 1 and 0.5 cfu/cm(2)were swabbed with a sponge, and the sponges were enriched for 18 h at 37 degrees C in universal pre-enrichment broth (UPB). Enriched broth cultures (EBC), cell pellets (CP), or phosphate-buffered saline-washed cell pellets (PBSCP) from enriched sponge samples were compared for detection of E. coli O157:H7, S. typhimurium DT104, or L. monocytogenes by the PCR using the BAX(TM)system. Recovery of the three organisms was effective for detection of each pathogen at initial levels of 10, 1 and 0.5 cfu/cm(2)when inoculated separately, or in combination, onto the beef samples. Use of EBC, CP, or PBSCP of sponge-swabbed samples eliminated problems associated with inhibition of the PCR by food components, time-consuming extraction of DNA, and inhibition due to large amounts of non-target DNA derived from the food. The procedure involving enrichment of sponge-swabbed beef samples in UPB followed by PCR amplification using EBC with the BAX system is the most efficient and simple method for detection of E. coli O157:H7, S. typhimurium DT104, and L. monocytogenes.

Animals↗

[Demonstration of enterotoxin in Staphylococcus aureus cultures by the ELISA test and the microslide test].

340 S. aureus strains, isolated either from routine food test samples, or from food leftovers, from healthy or sick persons in cases of suspected food poisoning in various places of the Federal Republic of Germany, were tested for enterotoxin production. The enterotoxin types A, B and C (SEA, SEB and SEC) were determined both by the ELISA and the microslide test (MS-test), the enterotoxin types D and E by the MS-test only. Comparison of the two test methods clearly shows that the ELISA technique is superior to the MS method: The sensitivity of the ELISA was at 2.5 ng, whereas that of the MS-test was limited to approximately 1 microgram. The ELISA revealed 6.2% more of the strains as enterotoxin producers, and reproducibility of the test results was also better with the ELISA than with the MS-test. Furthermore, the ELISA is far more efficient, consuming less test reagents, and with a capacity for testing more strains in a shorter time procedure. The incidence rate of the various enterotoxin types, referred to one representative strain each per outbreak of food poisoning or group of persons investigated, is as follows: 45.3% SEA, 17.2% SEB, 23.4% SEC. The incidence rate of SED, SEE or SE-type combinations was less than 5%. In sick persons, SEA- and SEC-producing strains occurred within the same range, i.e. 34.8% and 30.4% respectively. SEA-producing S. aureus strains had the highest frequency in food samples (61.5%) and in healthy individuals (53.6%).

Enterotoxins↗

Antigenicity of native and modified Kunitz soybean trypsin inhibitors.

Food provides a continuous antigenic stimulus to the immune system and the antigenicity of processed food proteins should be considered in toxicological evaluations. The antigenicity of the Kunitz trypsin inhibitor was studied using antibodies prepared by inoculating rabbits with native, heat-denatured, and N-acetylcysteine-treated Kunitz soybean trypsin inhibitors. Immunochemical studies using a competitive solid-phase enzyme immunoassay and two groups of sera revealed two patterns of antigenicity. Antibodies elicited with the denatured inhibitor were specific for the denatured conformation of the protein. In contrast, native inhibitor elicited antibodies that selectively recognized determinants in both native and heat-treated protein, but that did not bind trypsin inhibitors treated with N-acetylcysteine. These results imply that: the disulfide bonds must be intact to maintain the native antigenic conformation and the cysteine treatment may suppress allergic manifestations of soybean trypsin inhibitors and possibly other food proteins. These studies were extended by analyzing a panel of monoclonal antibodies prepared against native Kunitz trypsin inhibitor. The inhibitor has at least two distinct antigenic sites (epitopes), one of which is retained under denaturing conditions. The measurement of native Kunitz trypsin inhibitor in food samples by immunoassay appears practical. The relevance of these findings to food processing, food safety, and health is also discussed.

Animals↗

Survey of acrylamide in Turkish foods by an in-house validated LC-MS method.

A survey of retail Turkish foods was conducted for acrylamide using an in-house validated LC-MS method. The recoveries obtained for a variety of food matrices ranged between 81.2 and 96.8% for a spiking level of 500 ng g(-1). The limit of quantification was determined as 15-20 ng g(-1) depending on the food matrix in the basis of a signal-to-noise ratio of 5. A total of 120 food samples were analysed for acrylamide. The mean acrylamide levels in different food groups were in the order: crackers>potato chips>biscuits>cakes>baby foods>corn chips>cookies>breakfast cereals>breads>grilled vegetables>wafers>chocolates. The crumb of bread was free of acrylamide, whilst significant concentrations were found in the crust.

Acrylamide↗

Dietary benzo[a]pyrene intake and risk of colorectal adenoma.

We carried out a clinic-based case-control study specifically designed to address the hypothesis that dietary intake of polycyclic aromatic hydrocarbons (PAH) is associated with colorectal adenoma risk. We developed a food frequency questionnaire with detailed questions on meat-cooking methods and doneness levels and a benzo[a]pyrene (BaP) database (as a surrogate for total carcinogenic PAHs) based on the collection and analysis of a wide range of food samples. We estimated BaP intake derived from meat and from all foods to test its relationship with risk of colorectal adenomas. The median (10th and 90th percentiles) BaP intake in controls was 5 ng/d (0.2 and 66 ng/d) estimated from meat and 73 ng/d (35 and 140 ng/d) from all foods. In cases, median BaP intake was 17 ng/d (0.5 and 101 ng/d) from meat and 76 ng/d (44 and 163 ng/d) from all foods. Multivariate analysis was carried out on 146 cases and 228 controls. The odds ratios (95% confidence interval) for dietary BaP from meat with the first quintile as the reference group were 1.19 (0.51-2.80) for the second quintile, 1.71 (0.76-3.83) for the third quintile, 2.16 (0.96-4.86) for the fourth quintile, and 2.82 (1.24-6.43) for the fifth quintile (Ptrend=0.01). Increased risk of colorectal adenomas was more strongly associated with BaP intake estimated from all foods: 2.61 (1.08-6.29) for the second quintile, 4.21 (1.79-9.91) for the third quintile, 2.45 (0.98-6.12) for the fourth quintile, and 5.60 (2.20-14.20) for the fifth quintile (Ptrend=0.002). This study provides evidence that dietary BaP plays a role in colorectal adenoma etiology.

Adenoma↗

Identification and concentration of soy isoflavones in commercial cat foods.

OBJECTIVE: To determine the absolute and relative soy isoflavone content in commercial cat foods. SAMPLE POPULATION: 14 dry, 6 semimoist, and 22 moist commercial cat foods. PROCEDURE: Soy isoflavone content of each food was determined by use of acid-methanol hydrolysis and high-pressure liquid chromatography with ultraviolet absorbance detection. Isoflavones were identified and quantified by reference to authentic standards. RESULTS: Genistein and daidzein were the major soy isoflavones identified in 24 of 42 foods, with concentrations ranging from 1 to 163 microg/g of food. Foods labeled as containing soybean solids (16/42) had isoflavone concentrations > 11 microg/g. More dry (13/14) and semimoist (6/6) foods contained isoflavones than moist foods (5/22). Isoflavone content and food cost were negatively correlated for dry and semimoist foods but not for moist foods. Total amount of isoflavone consumed by cats fed these soy-containing foods as a sole maintenance diet was estimated to be between 0.6 and 4.5 mg/kg of body weight/d, which is comparable to concentrations in humans that result in a measurable although modest effect on serum concentrations of steroid and thyroid hormones. CONCLUSIONS AND CLINICAL RELEVANCE: Genistein and daidzein are common constituents of commercial cat foods. Predictors of isoflavone content included ingredient labeling, food type, and food cost. Soy isoflavones in some commercial cat foods were detected in amounts predicted to have a biological effect.

Animal Feed↗

Development of a solid-phase extraction method for determination of pheophorbide a and pyropheophorbide a in health foods by liquid chromatography.

A simple solid-phase extraction (SPE) method was developed for the liquid chromatography (LC) determination of pheophorbide (Phor) a and pyropheophorbide (Pyro) a in health foods such as chlorella, spirulina, etc. The food sample was extracted with 85% (v/v) acetone. The extract was acidified with hydrochloric acid and loaded on a C18 cartridge. After washing with water, Phor a and Pyro a were eluted with the LC mobile phase. Phor a and Pyro a were separated by isocratic reversed-phase LC and quantitated by fluorescence detection. The recoveries for spiked samples of chlorella and the extract were 87.1-102.0%. Commercial health foods (chlorella, spirulina, aloe, kale, Jews mallow, and green tea leaves) were analyzed using the SPE method. The values found for Phor a and Pyro a ranged from 2 to 788 microg/g and from <1 to 24 microg/g, respectively. There was no significant difference between the SPE method and the official method in Japan (spectrophotometry after liquid-liquid extraction). The advantages of the SPE method are the short extraction times, lack of emulsions, and reduced consumption of organic solvents compared with the official method in Japan. The SPE method is considered to be useful for the screening of Phor a and Pyro a in health foods.

Calibration↗