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PubMed2005

Identification and quantitation of species in complex DNA mixtures by real-time polymerase chain reaction.

Six TaqMan real-time polymerase chain reaction (PCR) systems using minor groove binding (MGB) probes have been developed for the detection quantitation of bovine, porcine, lamb, chicken, turkey, and ostrich DNA in complex samples. Species-specific amplification was achieved by combining only two fluorogenic probes and 10 oligonucleotide primers targeting mitochondrial sequences, decreasing the cost of the assay significantly. The limits of detection ranged from 0.03 to 0.80 pg of template DNA. Analysis of experimental mixtures containing two to four different species showed the suitability of the assay for detection of more than 1% of pork, chicken, or turkey and of more than 5% of cattle or lamb. The quantitation accuracy in samples containing 10-100% of beef or pork DNA was close to 90%. The system is complemented with one additional TaqMan MGB detector based on consensus sequence segments of the nuclear 18S ribosomal RNA gene. A method to evaluate the presence of unknown eukaryotic DNA in a mixture, where data derived from the species-specific detection are compared with the experimental values obtained from the general 18S detector, is presented. This method allows the validation of the quantitative measurements, providing an internal control of the total content of PCR-amplifiable DNA in the sample. The system was tested on DNA mixtures containing different shares of up to four different species and on DNA extracted from processed commercial food samples.

Animals↗

PubMed2004

Molecular and epidemiologic analysis of a county-wide outbreak caused by Salmonella enterica subsp. enterica serovar Enteritidis traced to a bakery.

BACKGROUND: An increase in the number of attendees due to acute gastroenteritis and fever was noted at one hospital emergency room in Taiwan over a seven-day period from July to August, 2001. Molecular and epidemiological surveys were performed to trace the possible source of infection. METHODS: An epidemiological investigation was undertaken to determine the cause of the outbreak. Stool and blood samples were collected according to standard protocols per Center for Disease Control, Taiwan. Typing of the Salmonella isolates from stool, blood, and food samples was performed with serotyping, antibiotypes, and pulsed field gel electrophoresis (PFGE) following XbaI restriction enzyme digestion. RESULTS: Comparison of the number of patients with and without acute gastroenteritis (506 and 4467, respectively) during the six weeks before the outbreak week revealed a significant increase in the number of patients during the outbreak week (162 and 942, respectively) (relative risk (RR): 1.44, 95% confidence interval (CI): 1.22-1.70, P value < 0.001). During the week of the outbreak, 34 of 162 patients with gastroenteritis were positive for Salmonella, and 28 of these 34 cases reported eating the same kind of bread. In total, 28 of 34 patients who ate this bread were positive for salmonella compared to only 6 of 128 people who did not eat this bread (RR: 17.6, 95%CI 7.9-39.0, P < 0.001). These breads were produced by the same bakery and were distributed to six different traditional Chinese markets., Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) was isolated from the stool samples of 28 of 32 individuals and from a recalled bread sample. All S. Enteritidis isolates were of the same antibiogram. PFGE typing revealed that all except two of the clinical isolates and the bread isolates were of the same DNA macrorestriction pattern. CONCLUSIONS: The egg-covered bread contaminated with S. Enteritidis was confirmed as the vehicle of infection. Alertness in the emergency room, surveillance by the microbiology laboratory, prompt and thorough investigation to trace the source of outbreaks, and institution of appropriate control measures provide effective control of community outbreaks.

Adolescent↗

PubMed1997

Impact of molecular biology on the detection of foodborne pathogens.

Molecular biological methods that use antibodies and nucleic acids to detect specific foodborne bacterial pathogens were scarcely known a decade and a half ago. Few scientists could have predicted that these tools of basic research would come to dominate the field of food diagnostics. Today, a large number of cleverly designed assay formats using these technologies are available commercially for the detection in foods of practically all major established pathogens and toxins, as well as of many emerging pathogens. These tests range from very simple antibody-bound latex agglutination assays to very sophisticated DNA amplification methods. Although molecular biological assays are more specific, sensitive, and faster than conventional (often cultural) microbiological methods, the complexities of food matrices continue to offer unique challenges that may preclude the direct application of these molecular biological methods. Consequently, a short cultural enrichment period is still required for food samples prior to analysis with these assays. The greater detection sensitivity of molecular biological methods may also affect existing microbiological specifications for foods; this undoubtedly will have repercussions on the regulatory agencies, food manufacturers, and also consumers.

Antibodies, Bacterial↗

PubMed1994

Mean daily intake of N-nitrosodimethylamine from foods and beverages in France in 1987-1992.

N-nitrosodimethylamine (NDMA) contamination was determined in 556 food samples and 75 beverages purchased in eastern France in 1987-1992. NDMA was found in 427 samples (68%). Data amassed enabled the first estimation to be made in France of the mean daily intake of NDMA from French foods and beverages in 1987-1992 (0.19 microgram/day). One-third of this daily exposure resulted from the consumption of alcoholic beverages.

Beverages↗

PubMed2006

Dietary PBDE intake: a market-basket study in Belgium.

A food market-basket, representative for the general Belgian population, containing various meat, fish and dairy food products, was assembled and analysed for its polybrominated diphenyl ether (PBDE) content. Additionally, fast food samples were also investigated. Based on the measured PBDE levels, an average daily dietary intake estimate of PBDEs was calculated. Of all foods analysed, fish had the highest average sum of PBDE levels (BDEs 28, 47, 99, 100, 153, 154, and 183; 460 pg/g ww), followed by dairy products and eggs (260 pg/g ww), fast food (86 pg/g ww) and meat products (70 pg/g ww). One fresh salmon filet had the highest total concentration of PBDEs (2360 pg/g ww), whereas levels in steak and chicken breast were the lowest of all foods analysed. BDE 209 was never found above LOQ in any food. PBDE intake calculations were based on the average daily food consumption in Belgium and were estimated between 23 and 48 ng/day of total PBDEs (lower and upper bound). This value is in accordance with what was previously reported for diets from geographical distinct areas, such as Canada, Finland, Spain, Sweden and the UK. Although it is only a minor constituent of the Belgian diet, fish is the major contributor to the total daily PBDE-intake (around 40%) due to the high PBDE levels in this type of food. Although low contaminated, meat products account for around 30% of the total dietary intake of PBDEs. Dairy products and eggs contribute to a lesser degree (less than 30%).

Animals↗

PubMed2006

Determination of 2-alkylcyclobutanones with electronic impact and chemical ionization gas chromatography/mass spectrometry (GC/MS) in irradiated foods.

The use of a column containing 60 g of silica gel for cleanup and the use of isobutane as a reactant reagent for chemical ionization-mass spectrometric analysis of the saturated and monounsaturated alkyl side-chain 2-alkylcyclobutanones (2-ACBs; specifically induced by irradiation from fat in foods until the proof of contrary) has improved both the sensibility and the selectivity of the method when applied for the detection of irradiated foods. The quality of the chromatograms obtained was improved, allowing the detection of food samples (avocados) irradiated at low doses (0.1 kGy) or irradiated ingredients included in low proportions (less than 5%, wt/wt) in nonirradiated culinary foods. These analytical modifications for the detection of 2-ACBs on the official EN 1785 method enable an extension of its current field of application using common equipment of food quality control laboratories.

Alkanes↗

PubMed1996

Periconceptional folate intake and neural tube defects.

Approximately 50% of neural tube defects may be folate-preventable and perhaps even more in other countries where prevalence is high. The Public Health Service has issued the recommendation that all women of childbearing age in the United States who are capable of becoming pregnant should consume 400 micrograms of folic acid/day for the purpose of reducing this risk. Ways in which a reproductive age woman could achieve this goal include: 1) fortifying a food sample with folic acid, 2) consuming supplements containing at least 400 mcg of folic acid, or 3) increasing nutrient intake by eating foods rich in folate. Advantages of consuming foods high in folate content are that it is a natural behavior and consistent with other dietary recommendations. However, this method is dependent upon a proper diet, and equivalencies of conjugated (dietary form) vs. unconjugated (in supplements) folate are unknown. The benefit of a supplementation policy is that the appropriate group can be targeted as pregnancies are planned, whereas primary limitations to taking a supplement would be compliance and most cases in need would not be reached. The advantage of fortification is that it is likely to reach everyone before conception, while the major disadvantage is that nontargeted populations will also receive more folic acid. Adequate consumption of folic acid should begin before and continue during at least the first 4 weeks after conception when the fetal neural tube is being formed. Standard methods of screening for neural tube defects should continue during pregnancy. The risk of a recurrent neural tube defect is 2-3%, and a higher periconception daily intake of folic acid (4 mg per day) is recommended.

Adolescent↗

PubMed1986

Improved immunological membrane filter method for detection of food-borne Salmonella strains.

An improved membrane filter method that involves the use of an enzyme-labeled antibody stain has been developed for the rapid detection of Salmonella species in foods. The procedure is carried out directly on a hydrophobic grid-membrane filter without requiring transfer by blotting to nitrocellulose. Pure cultures of 54 Salmonella species and 10 foods artificially contaminated with Salmonella colindale gave a positive reaction in which Salmonella colonies were visible as purple dots. Of 11 nonsalmonella organisms, only Citrobacter freundii reacted with Spicer-Edwards antiserum. Of 22 naturally contaminated food samples, 10 were positive for both the hydrophobic grid-membrane filter procedure of the Association of Official Analytical Chemists and the improved enzyme-labeled antibody stain method, and there was perfect agreement between the methods. Of these 10 positive samples, one was negative by the Health Protection Branch method; of the negative samples, two were positive by this latter method. The improved enzyme-labeled antibody stain method allows detection of Salmonella spp. in foods within 48 h, requires little equipment, and is inexpensive, easy to perform, and suitable for automated detection.

Citrobacter↗

PubMed1998

[Thiamine status of university teacher families in Changsha].

To evaluate the thiamine status of the population consuming the refined cereals and its products, we studied thiamine concentration in 23 food samples and human urine, and three-day dietary survey was conducted. The results showed that the thiamine content in the staple food was much lower than that in the raw material. The thiamine intake from diet was below to 80% RDA in 42.8% investigated families and the thiamine energy ratio was lower than 0.5 mg/4186.8 kJ in 71.5% of them. For the ratio of urinary tiamine (mg)/urinary inosine (g), 51.3% subjects exhibited decreased excretion, especially in the school-age children. We conclude that those people who take the refined cereals and its products as a staple food, is in the risk of thiamine deficiency.

Adolescent↗

PubMed1976

Collaborative study of the determination of sodium in dietetic foods by the sodium ion electrode method.

The dietetic food sample is diluted with total ionic strength adjusted buffer. The electrode potential of the sample is measured, known increments of sodium are added, and the electrode potential is measured after each addition. The mv readings obtained are plotted on Gran's paper and the concentration of sodium in the sample is found from the intercept of the extrapolated straight line. Seven processed dietetic vegetables and 2 spiked samples were sent to 7 collaborators. There was a 95.8% agreement between the results from the 7 collaborators and the Associate Referee for the unfortified samples. Recoveries of 98.5 and 95.1% were obtained on fortified samples, based on results from 5 collaborators. The statistical analysis indicates minimal bias and adequate precision for determination of sodium in dietetic food at the current regulatory requirements. The method has been adopted as official first action.

Diet, Sodium-Restricted↗

PubMed2002

Automated sample preparation by pressurized liquid extraction-solid-phase extraction for the liquid chromatographic-mass spectrometric investigation of polyphenols in the brewing process.

The analysis of polyphenols from solid plant or food samples usually requires laborious sample preparation. The liquid extraction of these compounds from the sample is compromised by apolar matrix interferences, an excess of which has to be eliminated prior to subsequent purification and separation. Applying pressurized liquid extraction to the extraction of polyphenols from hops, the use of different solvents sequentially can partly overcome these problems. Initial extraction with pentane eliminates hydrophobic compounds like hop resins and oils and enables the straightforward automated on-line solid-phase extraction as part of an optimized LC-MS analysis.

Automation↗

PubMed1993

Pesticide residue control in the years 1988-1989 in Italy.

The paper reports the monitoring for pesticide residue contamination of foodstuffs carried out in the years 1988-1989 by the Italian peripheral laboratories officially commissioned for the task. The tables show the analytical data for each class of substrate versus the limits in force at the time. 2506 food samples and 555 samples from the environment were analysed and a total of 16,259 determinations made. The residues were absent in 90% of the cases; the percentage of irregularities was 1.52%. These results confirm the data obtained in previous years, which is particularly significant considering the much larger number of analytical determinations performed this time. It has been deemed convenient to survey specific types of foodstuffs (for example, cereals and animal products) by examining a larger number of samples.

Edible Grain↗

PubMed2001

Analysis of dietary intake of selected metals in the NHEXAS-Maryland investigation.

As part of a large pilot investigation of multimedia exposure to several classes of environmental contaminants, the National Human Exposure Assessment Survey (NHEXAS)-Maryland study, we collected 388 semiquantitative food checklists and duplicate diet solid food samples, analyzed for arsenic, cadmium, chromium, and lead concentrations, from 80 individuals in Maryland in 1995-1996 in a repeated measures design. Here we explore several methods to infer foods most strongly associated with concentrations of these metals observed in the duplicate diet in our data set. We employed two techniques in which logarithmically transformed metal concentrations in the duplicate diet were regressed on individual food item consumption using algorithms designed to identify the foods most associated with the observed duplicate diet concentrations. We also employed an alternative strategy in which foods to be used as independent variables in regression were selected using data collected in national food consumption and residue surveys, with regression procedures proceeding with the selected foods in a similar manner. The concordance of foods selected as major predictors among these three techniques is noteworthy and is discussed. Finally, the Dietary Exposure Potential Model (DEPM) was used with the Dietary Checklist data to predict duplicate diet concentrations within our sample. A comparison between the predicted values and those observed gave R(2) values of 0.180, 0.206, and 0.076 for As, Cd, and Pb, respectively (p < 0.0001 in all cases). We discuss the significance of these observations and the implications for dietary-exposure-based risk analysis and dietary intake epidemiology.

Arsenic↗

PubMed1999

Cadmium and lead exposure associated with reduced growth rates, poorer fledging success of little blue heron chicks (Egretta caerulea) in south Louisiana wetlands.

Persistent contaminants in ecosystems are often monitored via organisms that bioaccumulate the pollutant of interest. Several published studies have used colonial wading birds to assess contamination by heavy metals in aquatic ecosystems. Using a nonlethal sampling regimen, this study identified chick feathers and guano as tissues that best reflect local contamination. Both cadmium and lead were detected in food samples (0.3-4.3 microg/g Cd, 0.8-9.3 microg/g Pb), guano (0.2-3.2 microg/g Cd, 1.0-9.5 microg/g Pb), and feathers (0.6-25.4 microg/g Cd, 1.2-16.9 microg/g Pb) from little blue heron chicks at levels similar to other studies. Chicks exposed to cadmium had significantly slower growth rates than nonexposed chicks, and exposure to lead was correlated with increased nestling mortality. The appropriate scale of comparison necessary to demonstrate these effects and the usefulness of wading birds as bioindicators are discussed.

Animals↗

PubMed1993

Occurrence and characteristics of Listeria in foods produced in Northern Ireland.

The incidence of L. monocytogenes and other Listeria species was determined for 513 food samples produced in Northern Ireland. Selected L. monocytogenes isolates were typed using MEE and RFLP analysis. The overall incidence of Listeria in the foods examined was 35% (Listeria monocytogenes 18.3%). The incidence of Listeria and L. monocytogenes in four different food categories (graded I-IV according to decreasing extent of processing) being: I, 11.1% Listeria and 4.7% L. monocytogenes; II, 27.1% Listeria and 12.2% L. monocytogenes; III, 89.1% Listeria and 50% L. monocytogenes and IV, 100% Listeria and 100% L. monocytogenes. Within food categories I and II, the incidence of Listeria on occasions varied markedly between similar products produced by different processors. The most frequently isolated species was L. innocua, followed by L. monocytogenes, L. seeligeri and L. welshimeri. The L. monocytogenes isolates were predominantly serogroup 1. A modified USDA method was the most productive of four enrichment procedures investigated. Over the one-year duration of the survey, a distinctive Listeria microflora could be discerned in products from certain processors and was confirmed in some cases by MEE and RFLP typing of L. monocytogenes isolates.

DNA, Bacterial↗

PubMed1985

An outbreak of food-borne enterotoxigenic Escherichia coli diarrhoea in England.

An outbreak of diarrhoea with abdominal pain occurred among members of the staff of a school and their guests after a social function at which a cold buffet was served. Sixty people attended the function and 43 subsequently completed questionnaires. Of these, 27 had diarrhoea. The median incubation period was 36 h and the range 12-66 h. Food history analysis showed an association between diarrhoea and eating curried turkey mayonnaise. Stool specimens from 13 of those who developed diarrhoea were examined: Escherichia coli 06.H16 (producing heat-stable and heat-labile enterotoxins) was found in nine specimens and E. coli 027.H20 (producing heat-stable enterotoxin) in 11 specimens. Eight patients had both strains and only one was negative for enterotoxigenic E. coli. Food samples were not available for examination. Enterotoxigenic E. coli should be considered as a possible cause in well-defined outbreaks of food-borne diarrhoea with abdominal pain.

Abdomen↗

PubMed2002

The fluoride content of foods and beverages from negligibly and optimally fluoridated communities.

In the spring of 1996, foods and beverages most commonly consumed by adolescents were analyzed for fluoride as part of a larger investigation. These foods were selected by interviewing 711 adolescents, 12-14 years of age, who were long-time residents of either an optimally or negligibly fluoridated community. The brand names of the identified foods and beverages most commonly purchased were determined by interviews with the parents. A total of 441 brand-name food and beverage items were purchased from both communities and were individually analyzed for fluoride. These analyses were done in order to estimate the fluoride content of various kinds of foods and beverages and to determine whether or not there was a significant difference between the two communities in the amount of fluoride ingested from these dietary sources. The food and beverage items were classified into dietary groups based on US Department of Agriculture (USDA) guidelines. Overall, the fluoride content of the sampled foods and beverages was low. In addition, there was no significant difference in the fluoride content of the same pre-packaged or ready-to-eat food or beverage items purchased in the two communities. However, a significant difference was found between the two communities in the fluoride content of fountain beverages and in cooked or reconstituted foods prepared using local water from the respective communities. Based on these results, we have estimated the mean daily, dietary fluoride intake for 3-5-year-old children who are more susceptible to developing dental fluorosis.

Adolescent↗

PubMed2002

Food and macronutrient intake of male adolescent Kalenjin runners in Kenya.

A nutritional survey based on twelve adolescent male Kalenjin runners in Kenya during a 2-week field study was carried out in order to determine the composition of their diet and make a comparison with macronutrient recommendations for athletes. Food samples were collected for analysis of macronutrient distribution and energy content from main meals and the macronutrient distribution and energy content of additional food intake were based on the information of a 24 h recall interview and estimated from food tables. The diet of the Kalenjin runners was very high in carbohydrate (71 % 8.7 g/kg body weight per d) and very low in fat (15 %). Intake of total protein (13 %; 1.6 g/kg body weight per d) was above the daily intake recommended by the Food and Agriculture Organization/World Health Organization/United Nations University (FAO/WHO/UNU), while essential amino acid intake was estimated to be in the borderline-to-low range based on FAO/WHO/UNU recommendations for children <12 years and adults. The energy intake was mainly derived from vegetable sources (90 %) with maize and kidney beans as the staple food (81 %). The diet of the Kalenjin runners met recommendations for endurance athletes for total protein and most essential amino acid intake as well as carbohydrate intake even though it was based on a small range of food items.

Adolescent↗