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Where we are in retail food safety, how we got to where we are, and how do we get there?

Food safety has not yet been attained. This is evident from reported foodborne-disease outbreaks, laboratory-confirmed cases of diseases that can be foodborne, estimates of foodborne illness based on surveillance data, and out-of-compliance risk factors. Several activities have had an impact on food safety, but there are limitations in the way each of those activities has been or is being conducted. The activities include foodborne-disease surveillance; food sampling and testing; swabbing and testing of utensils; inspection and enforcement of regulations; use of the Food Code; on-site hazard analyses, on-site monitoring of critical control points and prompt corrective actions; applied research and challenge testing; training of public-health and food regulatory personnel; training of food workers, supervisors, and managers; and education of the public. To attain food safety, we must use common (microbiological) sense and understand the principles of transmission of foodborne-disease etiological agents and their control. A change of attitudes and program focus is necessary.

Consumer Product Safety↗

An outbreak of Campylobacter jejuni associated with consumption of chicken, Copenhagen, 2005.

In May/June 2005 an outbreak of diarrhoeal illness occurred among company employees in Copenhagen. Cases were reported from seven of eight companies that received food from the same catering kitchen. Stool specimens from three patients from two companies were positive for Campylobacter jejuni. We performed a retrospective cohort study among employees exposed to canteen food in the three largest companies to identify the source of the outbreak and to prevent further spread. Using self-administered questionnaires we collected information on disease, days of canteen food eaten and food items consumed. The catering kitchen was inspected and food samples were taken. Questionnaires were returned by 295/348 (85%) employees. Of 247 employees who ate canteen food, 79 were cases, and the attack rate (AR) was 32%. Consuming canteen food on 25 May was associated with illness (AR 75/204, RR=3.2, 95%CI 1.3-8.2). Consumption of chicken salad on this day, but not other types of food, was associated with illness (AR=43/97, RR=2.3, 95%CI 1.3-4.1). Interviews with kitchen staff indicated the likelihood of cross-contamination from raw chicken to the chicken salad during storage. This is the first recognised major Campylobacter outbreak associated with contaminated chicken documented in Denmark. It is plausible that food handling practices contributed to transmission, and awareness of safe food handling and storage has since been raised among kitchen staff. The low number of positive specimens accrued in this outbreak suggests a general underascertainment of adult cases in the laboratory reporting system by a factor of 20.

Animals↗

Prevalence and molecular characterization of Aeromonas spp. in ready-to-eat foods in Italy.

A survey was carried out in Italy to ascertain the prevalence of Aeromonas spp. in ready-to-eat foods (vegetables, cheeses, meat products, and ice creams) and the level of molecular heterogeneity of the isolates found by macrorestriction analysis of genomic DNA with pulsed-field gel electrophoresis (PFGE). In total, 46 (14.4%) of the 320 food samples examined were found positive for Aeromonas spp. The highest percentages of isolation were discovered in vegetables, particularly lettuce (45.0%), endive (40.0%), and rucola (20.0%). Ricotta was the only cheese type analyzed that showed a high frequency of isolation (45.0%). Among meat products, salami and raw ham (25.0% of samples positive) and, to a lesser extent, baloney (5.0%) were found positive for Aeromonas spp. Aeromonas hydrophila was the most common isolate from foods of animal origin, whereas Aeromonas caviae was the dominant species in vegetables. No motile aeromonads were found in ice cream samples. Aeromonas isolates showed a high level of genetic heterogeneity, because 24 PFGE patterns were identified among 27 A. hydrophila strains and 20 PFGE patterns were found in 23 A. caviae isolates. In conclusion, consumers of ready-to-eat foods in Italy are regularly exposed to many genetically distinct strains of A. hydrophila and A. caviae without evident signs of malaise, and therefore, few of these strains, if any, are likely to be pathogenic.

Aeromonas↗

Rapid, sensitive assay for staphylococcal enterotoxin and a comparison of serological methods.

Reversed passive hemagglutination was used to assay enterotoxin in culture filtrates and in food samples. With cells tanned and then sensitized with antitoxin globulin and preserved with either formaldehyde or pyruvic aldehyde, as little as 0.0007 mug of enterotoxin was detectable. The results of hemagglutination tests compared well with those obtained by quantitative precipitin tests or by immunodiffusion, but hemagglutination was 50 to 100 times more sensitive than the immunodiffusion technique. In addition, results of the hemagglutination test were available within a few hours, and neither elimination of interfering proteins from food extracts nor concentration of the sample, both of which are necessary for immunodiffusion, was required for this procedure.

Antitoxins↗

Determination of polybrominated diphenyl ethers and polybrominated dibenzo-p-dioxins/dibenzofurans in marine products.

Polybrominated diphenyl ethers (PBDEs) have been widely used as flame retardants in plastics and textile coatings, and these compounds have been recognized as ubiquitous environmental contaminants. Furthermore, it is considered a serious problem that polybrominated dibenzo-p-dioxins and dibenzofurans (PBDD/DFs), having toxicities similar to those of chlorinated dioxins, are generated by the manufacture of brominated flame retardants (BFRs) such as PBDEs, and formed by the combustion of substances containing BFRs. Several congeners of PBDD/DFs and PBDEs have been detected in the adipose tissue of the Japanese. Although food is suspected as an exposure source, little information is available regarding the levels of these brominated compounds in food, as compared with information regarding dioxin or polychlorinated biphenyls. It is necessary to investigate the levels of these brominated organic compounds in various foods and to estimate their influence in the case of human exposure. We developed an efficient method of analyzing PBDEs and PBDD/DFs contents in food samples using accelerated solvent extraction and determined the concentrations in several marine products such as raw fish, processed foods, and seaweed purchased in Japan. A recovery test (n = 5) using the method and involving dried fish showed acceptable recoveries of 57.7-78.5% (RSD 5.4-15.9%) for PBDEs and 50.0-56.4% (RSD 1.5-7.9%) for PBDD/DFs. In the analysis of marine product samples, several congeners of PBDEs were detected in raw fish, processed fish, and seaweed; the highest concentration of sigmaPBDEs was detected in yellowtail (1161 pg/g whole basis), followed by mackerel (553.5 pg/g whole basis). The most dominant congener present in these marine samples was 2,2',4,4'-tetraBDE (#47).

Animals↗

Menadione-catalyzed luminol chemiluminescence assay for the rapid detection of viable bacteria in foods under aerobic conditions.

A menadione-catalyzed luminol chemiluminescence assay was developed for the rapid detection and estimation of viable bacteria in foods. The principle of this assay is based on the extracellular menadione-catalyzed active oxygen spieces (O2- and H2O2) generated by the activity of NAD(P)H:menadione oxidoreductase in viable cells. This luminol chemiluminescence assay requires 10 min for the incubation of cells with menadione and then 2 s for the measurement of chemiluminescence intensity after an injection of luminol solution without the treatment of cell lysis. This method was evaluated using liquid food samples of milk, vegetable juice, green tea, and coffee spiked with Escherichia coli ATCC 25922. The study result revealed that E. coli contamination at 1 to 10 CFU/ml in these foods could be detected after incubation at 37 degrees C for 7 h in an enrichment medium; however, the green tea and coffee samples requires filtration. This method could be a useful tool for the rapid evaluation of microbial food contamination.

Colony Count, Microbial↗

Detection of Salmonella enterica serovar Typhi (S. Typhi) by selective amplification of invA, viaB, fliC-d and prt genes by polymerase chain reaction in mutiplex format.

AIMS: Development of a PCR assay that can target multiple genes for rapid detection of Salmonella enterica serovar Typhi (S. Typhi) from water and food samples. METHODS AND RESULTS: PCR primers for invasion, O, H and Vi antigen genes, invA, prt, fliC-d and viaB were designed and used for the rapid detection of S. Typhi by multiplex PCR. Internal amplification control, which co-amplified with prt primers, was also included in the assay. The results showed that all cultures of Salmonella were accurately identified by the assay with no nonspecific amplification in other cultures. The assay had 100% detection probability when a cell suspension of 10(4) CFU ml(-1) (500 CFU per reaction) was used. Salmonella Typhi bacteria were artificially inoculated in the water and food (milk and meat rinse) samples and detected by mPCR after overnight pre-enrichment in buffered peptone water. No Salmonella bacteria could be detected from water samples collected from the field by mPCR or standard culture method. CONCLUSIONS: The developed mPCR assay provides specific detection of S. Typhi. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid methods for detection of S. Typhi from complex environmental matrices are almost nonexistent. The mPCR assay reported in this study can be useful to identify S. Typhi bacteria in field environmental samples.

Antigens, Bacterial↗

Dietary intake of hexachlorobenzene in Catalonia, Spain.

To assess the dietary intake of hexachlorobenzene (HCB) by the population of Catalonia, Spain, a total-diet study was carried out. Concentrations of HCB were determined in food samples randomly acquired in seven cities of Catalonia between June and August 2000. A total of 11 food groups were included in the study. HCB levels were determined by HRGC/HRMS. Estimates of average daily food consumption were obtained from recent studies. HCB intake was estimated for five population groups: children (aged 4 to 9 years), adolescents (aged 10 to 19 years), male and female adults (aged 20 to 65 years), and seniors (aged >65 years). In general, HCB residues in foods were rather low excepting dairy products with a mean concentration of 0.869 ng/g wet weight. Total dietary intakes of HCB (microgram per kilogram body weight/day) were the following: children (0.0064), adolescents (0.0031), female adults (0.0025), male adults (0.0024) and seniors (0.0019). All these values are considerably lower than the WHO tolerable daily intake (TDI), which is 0.17 microg kg(-1) day(-1) for non-cancer effects and 0.16 microg kg(-1) day(-1) for neoplastic effects in humans.

Adolescent↗

Concentrations of arsenic, cadmium, mercury, and lead in common foods and estimated daily intake by children, adolescents, adults, and seniors of Catalonia, Spain.

This study was designed to estimate the dietary intake of arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb) by the general population of Catalonia, Spain. The concentrations of these elements were determined in food samples randomly acquired in seven cities of Catalonia between June and August 2000. A total of 11 food groups were included in the study. As, Cd, Hg, and Pb levels were measured by ICP-MS and AAS. The dietary intake of these elements was determined by a total diet study. Calculations were carried out on the basis of recent data on the consumption of the selected food items. Trace element intake was estimated for five population groups: children, adolescents, male and female adults, and seniors. The highest dietary intakes of As (223.6 microg/day), Cd (15.7 microg/day), Hg (21.2 microg/day), and Pb (28.4 microg/day) corresponded to male adults. For all analyzed elements, fish and shellfish was the group showing the highest contribution to the respective intakes. In comparison with previous results, a general decrease in As, Cd, Hg, and Pb intake has occurred. The dietary intake of these elements was also compared with the provisional tolerable weekly intake (PTWI). Dietary intakes of As, Cd, Hg, and Pb by the population of Catalonia are currently well below the respective PTWIs.

Adolescent↗

Critical control points of complementary food preparation and handling in eastern Nigeria.

OBJECTIVE: To investigate microbial contamination and critical control points (CCPs) in the preparation and handling of complementary foods in 120 households in Imo state, Nigeria. METHODS: The Hazard Analysis Critical Control Point (HACCP) approach was used to investigate processes and procedures that contributed to microbial contamination, growth and survival, and to identify points where controls could be applied to prevent or eliminate these microbiological hazards or reduce them to acceptable levels. Food samples were collected and tested microbiologically at different stages of preparation and handling. FINDINGS: During cooking, all foods attained temperatures capable of destroying vegetative forms of food-borne pathogens. However, the risk of contamination increased by storage of food at ambient temperature, by using insufficiently high temperatures to reheat the food, and by adding contaminated ingredients such as dried ground crayfish and soybean powder at stages where no further heat treatment was applied. The purchasing of contaminated raw foodstuffs and ingredients, particularly raw akamu, from vendors in open markets is also a CCP. CONCLUSION: Although an unsafe environment poses many hazards for children's food, the hygienic quality of prepared food can be assured if basic food safety principles are observed. When many factors contribute to food contamination, identification of CCPs becomes particularly important and can facilitate appropriate targeting of resources and prevention efforts.

Child↗

Effects of food texture and sample thickness on mandibular movement and hardness assessment during biting in man.

This study was designed to investigate the relationship among jaw movements, physical characteristics of food, and sensory perception of hardness in man. Vertical movements of the mandible were recorded with an infrared tracking device in humans during biting on two test foods, carrot and cheese. Samples of standard length (2 cm) and width (2 cm) were prepared in three different thicknesses (0.5, 1.0, and 1.5 cm). Nine subjects were asked to perform two types of bite with their incisor teeth. In the first, they cut through the food, then stopped and spat out the pieces (bite alone); in the second, biting was followed by mastication and swallowing (bite+chew). The 12 conditions (thickness x3, food x2, and bite x2) were presented in a random order within each block, and blocks were repeated five times (60 trials per subject). Subjects also estimated the hardness of the samples twice for each condition on visual analogue scales (VAS) 100 mm long. The duration, vertical amplitude, and maximum vertical velocity of the mandible during biting were calculated by computer for the three phases of the movements (opening, and fast and slow closing). Multilevel statistical models were used for data analysis. The estimated hardness scores associated with the first bite of thin carrot (59.0 VAS units) was significantly greater than for cheese (16.8 VAS units). The type of bite had no significant effect on these scores, but the estimate of hardness was significantly greater for the thickest sample (+13.3 VAS units). Food type had its strongest effect on the slow-closing phase. In particular, the peak velocity that followed the fracturing of the food sample was much greater for carrot than for cheese (thin, 34.1 mm.s-1 vs. 26.6 mm.s-1), and the difference between foods increased with thickness. The amplitude of opening was significantly greater for the thickest sample than for the other two. There were no significant relationships between VAS scores and the movement parameters. These results suggest that, when humans bite food: (1) changing the thickness of food has a greater effect on movement parameters than changing from soft to hard food, (2) the parameters of biting change little if biting is followed by mastication, (3) hardness perception is dependent on the thickness of food, (4) hardness perception is not different when food is removed from the mouth than when it is chewed and swallowed, and (5) there is no relationship between any of the parameters of movement that change with food type and the perceived hardness of food.

Adult↗

An ultracentrifugation-based approach to the detection of hepatitis A virus in soft fruits.

A method was developed for detection of hepatitis A virus (HAV) in soft fruits (raspberries and strawberries). After washing the sample in 1 M sodium bicarbonate with added soya protein, fruits were removed by slow speed centrifugation, then particulate material and residual pectin were removed from the supernatant by flocculation and pectinase treatment during another slow speed centrifugation. Virus particles were then sedimented by ultracentrifugation. RNA was extracted from the virus particles, and nested RTPCR was performed on the nucleic acid extract. Nested RTPCR comprised an RTPCR, followed by PCR to amplify sequences within the amplicon. Internal amplification controls (IACs) were constructed for both the RTPCR and the PCR. The sensitivity of the nested RTPCR was approximately 10 RTPCRU. The overall method was shown to be able to detect 10(4) RTPCRU HAV in 90 g fresh strawberries, and 10(3) RTPCRU HAV in 60 g fresh raspberries. It is estimated that the lowest possible limit of detection of the method should be between 40 and 400 RTPCRU HAV per fruit sample. The method can be performed within one day, in suitably equipped microbiological laboratories, and is suitable for routine screening of food samples, and for analysis of suspected samples, e.g. during outbreak investigations.

Centrifugation↗

Search for B-1 aflatoxin and similar fluorescent compounds in 'misso'.

Following previous studies on the investigation of B-1 Aflatoxin in food samples (4,5) we report in this paper the results obtained using thin layer chromatography (TLC) with three different solvent systems and spectrophotometric analysis in order to differentiate actual B-1 Aflatoxin and Aflatoxin-like substances. Twenty 'Misso' samples were studied and we could detect Aflatoxin in 3 of them. An Aflatoxin-suspected substance was also isolated. No Aflatoxin-producing Aspergillus was isolated from the samples analysed.

Aflatoxins↗

The specificity of satiety: the influence of foods of different macronutrient content on the development of satiety.

The effects of consuming equicaloric preloads with different macronutrient contents on the development of satiety were investigated. Ten normal-weight, nondieting women fasted overnight and then rated hunger, fullness, and food preferences, and the pleasantness of the taste of food samples before and after five different preloads. The greatest changes in the pleasantness of the taste of the food occurred for the eaten food relative to the uneaten food. Macronutrient content of the preloads did not significantly influence the magnitude of these changes or the pleasantness of foods with similar nutrient contents. Therefore, no evidence of nutrient-specificity was obtained. Nutrient composition had a differential effect on hunger, fullness and food intake. Ratings of hunger decreased and fullness increased following the high starch and high protein preloads to a greater extent than after the high fat, high sucrose and mixed content preloads. When a self-selection meal was offered 2 hours after the preloads energy intake and preference for high carbohydrate and high fat food items were significantly decreased by the high protein and high starch preloads. However, no specific reduction in macronutrient intake was observed. In conclusion, differential effects of the preloads were observed in subjective ratings of hunger, fullness, preferences and subsequent food intake, but there was no indication that satiety was macronutrient-specific.

Adult↗

Oligonucleotide microarray for identification of Enterococcus species.

For detection of most members of the Enterococcaceae, the specificity of a novel oligonucleotide microarray (ECC-PhyloChip) consisting of 41 hierarchically nested 16S or 23S rRNA gene-targeted probes was evaluated with 23 pure cultures (including 19 Enterococcus species). Target nucleic acids were prepared by PCR amplification of a 4.5-kb DNA fragment containing large parts of the 16S and 23S rRNA genes and were subsequently labeled fluorescently by random priming. Each tested member of the Enterococcaceae was correctly identified on the basis of its unique microarray hybridization pattern. The evaluated ECC-PhyloChip was successfully applied for identification of Enterococcus faecium and Enterococcus faecalis in artificially contaminated milk samples demonstrating the utility of the ECC-PhyloChip for parallel identification and differentiation of Enterococcus species in food samples.

DNA, Bacterial↗

Antibiotic resistance of retail food and human Campylobacter isolates on the island of Ireland from 2001-2002.

The antimicrobial resistance profiles of Campylobacter isolates recovered from a range of retail food samples (n=374) and humans (n=314) to eight antimicrobial compounds were investigated. High levels of resistance in food C. jejuni isolates were observed for ceftiofur (58%), ampicillin (25%) and nalidixic acid (17%) with lower levels observed for streptomycin (7.9%) and chloramphenicol (8.3%). A total of 80% of human C. jejuni isolates were resistant to ceftiofur, while 17% showed resistance to ampicillin and nalidixic acid, 8.6% to streptomycin and 4.1% to chloramphenicol. Resistance to clinically relevant antimicrobials such as erythromycin, ciprofloxacin and tetracycline was 6.7, 12, and 15% respectively for all food isolates and was similar to corresponding resistance prevalences observed for human isolates, where 6.4, 12 and 13% respectively were found to be resistant. Comparisons of C. jejuni isolates in each location showed a high degree of similarity although some regional variations did exist. Comparison of total C. jejuni and C. coli populations showed minor differences, with C. jejuni isolates more resistant to ampicillin and ceftiofur. Multidrug resistance patterns showed some profiles common to human and clinical isolates.

Anti-Bacterial Agents↗

Fluorescence polarization immunoassays for pesticides.

Fluorescence polarization immunoassay methods for the detection of pesticides and their metabolites or degradation products are reviewed. Advantages and limitations for application to pesticide detection in environmental and food samples are discussed. The influence of the structure of fluorescent-labeled tracers and the affinity and specificity of antibodies on analytical performance is examined. The methods are simple, readily automated, and rapid (total time for assay of a water sample is about 1 min) with sensitivity of 1 - 10 ng/ml pesticide in 0.01 - 0.1 ml sample.

Environmental Pollutants↗

A review of the significance of animal food products as potential pathways of human exposures to dioxins.

The polychlorinated dibenzo-p-dioxins and dibenzofurans (dioxins) are groups of compounds with similar chemical and toxicological properties. Carcinogenicity was considered the most serious toxic end point when setting previous regulatory policies, but recent concerns have focused on the possible endocrine-disrupting activities of the dioxins. Toxicity is related to the 2,3,7,8 pattern of chlorine substitution, a pattern that also leads to chemical and metabolic stability. Dioxins are practically insoluble in water and concentrate in lipids of biological systems, leading to low background concentrations in fat of the general human population. Major environmental sources of dioxins are emissions from industrial chlorination processes and combustion of materials containing chlorine. Inhalation and water have been ruled out as significant exposure pathways, which suggests that food is the primary source. Pathways of entry into food chains are atmospheric transport of emissions and their subsequent deposition on plants, soils, and water. The major food sources seem to be fat-containing animal products and some seafoods. This conclusion is based on evaluations of potential environmental pathways involving dioxins and related compounds. Generally, dioxins and other lipophilic compounds are not taken up and translocated by plants, so residues in foods and feeds derived from seeds should be negligible. Animals on high-roughage diets, or those that ingest contaminated soil, are the most likely to accumulate dioxin residues from the environment. The conclusion that animal products are a major source of human exposure requires verification by appropriate food sampling programs and animal metabolism studies. If it is desirable to reduce human exposure to dioxins via the food supply, reduction of sources would be a more effective strategy than changing agricultural practices and food consumption patterns.

Animal Feed↗