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Botulism.

Botulism is a rare disease with 4 naturally occurring syndromes: foodborne botulism is caused by ingestion of foods contaminated with botulinum toxin, wound botulism is caused by Clostridium botulinum colonization of a wound and in situ toxin production, infant botulism is caused by intestinal colonization and toxin production, and adult intestinal toxemia botulism is an even rarer form of intestinal colonization and toxin production in adults. Inhalational botulism could result from aerosolization of botulinum toxin, and iatrogenic botulism can result from injection of toxin. All forms of botulism produce the same distinct clinical syndrome of symmetrical cranial nerve palsies followed by descending, symmetric flaccid paralysis of voluntary muscles, which may progress to respiratory compromise and death. The mainstays of therapy are meticulous intensive care (including mechanical ventilation, when necessary) and timely treatment with antitoxin.

Botulinum Antitoxin↗

Risk from inhaled mycotoxins in indoor office and residential environments.

Mycotoxins are known to produce veterinary and human diseases when consumed with contaminated foods. Mycotoxins have also been proposed to cause adverse human health effects after inhalation exposure to mold in indoor residential, school, and office environments. Epidemiologic evidence has been inadequate to establish a causal relationship between indoor mold and nonallergic, toxigenic health effects. In this article, the authors model a maximum possible dose of mycotoxins that could be inhaled in 24 h of continuous exposure to a high concentration of mold spores containing the maximum reported concentration of aflatoxins B1 and B2, satratoxins G and H, fumitremorgens B and C, verruculogen, and trichoverrols A and B. These calculated doses are compared to effects data for the same mycotoxins. None of the maximum doses modeled were sufficiently high to cause any adverse effect. The model illustrates the inefficiency of delivery of mycotoxins via inhalation of mold spores, and suggests that the lack of association between mold exposure and mycotoxicoses in indoor environments is due to a requirement for extremely high airborne spore levels and extended periods of exposure to elicit a response. This model is further evidence that human mycotoxicoses are implausible following inhalation exposure to mycotoxins in mold-contaminated home, school, or office environments.

Adult↗

Risk Behavior for Travelers' Diarrhea Among Finnish Travelers.

Background: Contaminated food and drink are the primary sources of traveler's diarrhea (TD). Identification of the characteristics that make a traveler more prone to TD is needed to improve prevention and therapy of this illness. Methods: We evaluated, by questionnaire, the association of dietary errors with TD among 933 adult Finnish tourists vacationing in Morocco. A stool specimen was obtained from patients in the group that developed TD at the onset of the diarrheal episode, and from all participants in the study on their return to Finland. Results: Only 5% of the 933 subjects who responded to the questionnaire, and who gave a stool sample, had adhered strictly to generally accepted recommendations. About 45% made five or more dietary errors during the trip; of these, 75% consumed raw vegetables, 65% salads, 33% puddings, 32% mayonnaise or other cold dressings, 31% consumed food bought from street vendors, 29% consumed cold meat, 18% cold sandwiches, 4% drank tap water, and 2% consumed raw meat. The age of the subjects did not correlate with the number of dietary errors. Subjects who had been abroad during the preceding 12 months committed more dietary errors than those who had not (mean 3.9 versus 4.5; p <.001). Also, subjects who spent 2 weeks in Morocco committed more dietary errors than those who spent only 1 week (mean 4.0 versus 4.8; p <.001). However, no association between TD and the number of dietary errors was observed. Differences with respect to eating habits were not observed between subjects, with or without TD or with or without an identified pathogen in stool specimens. There was no correlation between eating habits and the presence, or absence, of a specific pathogen in the stool sample; this is with the exception of Campylobacter spp: subjects with this pathogen consumed steak tartar or salad more often than did other subjects. Conclusions: Etiologic agents are so ubiquitous in this high-risk area that instructions to avoid certain foodstuffs commonly thought to be contaminated may be to some extent without value. Moreover, dietary self restraint proved to be impossible in the real situation. (J Travel Med 2:77-84, 1995)

Journal Article↗

New reversed-phase liquid chromatographic method to detect aflatoxins in food and feed with cyclodextrins as fluorescence enhancers added to the eluent.

The effect of succynil-beta-cyclodextrin (beta-CD-Su), dimethyl-beta-cyclodextrin (DIMEB) and beta-cyclodextrin (beta-CD) on the fluorescence of aflatoxins B1, B2, G1, G2 and M1 (AFB1, AFB2, AFG1, AFG2 and AFM1) was studied: beta-CD-Su promoted the largest fluorescence enhancement for AFB1 and AFM1 while DIMEB showed better results for AFG1 . On the basis of the fluorescence enhancement, a new RP-HPLC method for detecting aflatoxins B1, B2, G1, G2 and M1 was developed using cyclodextrins directly dissolved in the LC eluent. Aflatoxins B1, B2, G1 and G2 were resolved using a MICRA NPS ODS-1 column using methanol-water as mobile phase to which 6 x 10(-3) M beta-CD-Su or beta-CD were added. Chromatographic responses of AFB1 and AFG1 achieved using beta-CD dissolved in the mobile phase were enhanced, respectively, 8 and 12 times, and 10 and 15 times with beta-CD-Su. Detection limits lower than 0.3 microg/kg were achieved for all the four aflatoxins. Aflatoxin M1 was analysed using a Spherisorb S3 ODS-2 Narrow Bore column and methanol-water as mobile phase with added 2 x 10(-3) M beta-CD-Su. An area enhancement of 1.5 was detected for the toxin and the detection limit achieved under these analytical conditions was lower than 0.0005 microg/kg. Both methods were statistically validated showing a linear response for all the aflatoxins tested (R2 > or = 0.99), and applied to the analysis of spiked and naturally contaminated food samples.

Aflatoxins↗

Construction and analysis of fractional multifactorial designs to study attachment strength and transfer of Listeria monocytogenes from pure or mixed biofilms after contact with a solid model food.

The aim of this study was to establish which of seven factors influence the adhesion strength and hence bacterial transfer between biofilms containing Listeria monocytogenes (pure and two-species biofilms) and tryptone soya agar (TSA) as a solid organic surface. The two-species biofilms were made of L. monocytogenes and one of the following species of bacteria: the nonpathogenic organisms Kocuria varians, Pseudomonas fluorescens, and Staphylococcus sciuri and CCL 63, an unidentified gram-negative bacterium isolated from the processing plant environment. We used biofilms prepared under conditions simulating open surfaces in meat-processing sites. The biofilm's adhesion strength and population were evaluated by making 12 contacts on a given whole biofilm (4.5 cm(2)), using a new slice of a sterilized TSA cylinder for each contact, and plotting the logarithm CFU . cm(-2) detached by each contact against the contact number. Three types of detachment kinetics were observed: biphasic kinetics, where the first slope may be either positive or negative, and monophasic kinetics. The bacteria that resisted a chlorinated alkaline product and a glutaraldehyde- and quaternary ammonium-based disinfectant had greater adhesion strengths than those determined for untreated biofilms. One of the four non-Listeria strains studied, Kocuria varians CCL 56, favored both the attachment and detachment of L. monocytogenes. The stainless steel had smaller bacterial populations than polymer materials, and non-Listeria bacteria adhered to it less strongly. Our results helped to evaluate measures aimed at controlling the immediate risk, linked to the presence of a large number of CFU in a foodstuff, and the delayed risk, linked to the persistence of L. monocytogenes and the occurrence of slightly contaminated foods that may become dangerous if L. monocytogenes multiplies during storage. Cleaning and disinfection reduce the immediate risk, while reducing the delayed risk should be achieved by lowering the adhesion strength, which the sanitizers used here cannot do at low concentrations.

Agar↗

The protective effect of some food ingredients on Staphylococcus aureus MF31.

The upper limiting temperature of growth of Staphylococcus aureus MF31 in heart infusion broth (HI) was about 44 degrees C but addition of monosodium glutamate (MSG) and soy sauce permitted the organism to grow above this temperature. This effect is similar to that of NaCl. Tomato ketchup, Worcestershire and HP sauces added to HI did not allow growth at the non-permissive temperature of 46 degrees C but death was delayed. Staphylococcus aureus died in unsupplemented chicken meat slurry at 46 degrees C but grew at 48 degrees C in slurry supplemented with 5.8% NaCl and survived incubation for 18 h at 50 degrees C in slurry supplemented with 5.8% NaCl and 5% MSG. Cultures grown at 37 degrees C had a D60 value of 2 min in 50 mmol/l Tris (pH 7.2) buffer. Cultures grown at 46 degrees C in HI containing 5.8% NaCl had a D60 value of 8 min in Tris buffer. Addition of 5.8% NaCl plus 5% MSG to the buffer increased the D60 by a factor of about 7 for both cultures. In storage experiments at room temperature, the culture grown at 37 degrees C and at 46 degrees C plus 5.8% NaCl died at about the same rate in salami. In milk powder, however, the count of 37 degrees C culture decreased from 10% g to 10(6)/g in 5 weeks while the count of 46 degrees C culture remained unchanged. In cottage cheese, freeze-dried rice and macaroni, the 37 degrees C cultures also died more rapidly. It is suggested that cultures grown at 46 degrees C plus 5.8% NaCl may be suitable for experiments with artificially contaminated foods.

Cheese↗

Evaluation of certain food additives and contaminants.

This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, with a view to recommending acceptable daily intakes (ADIs) and to prepare specifications for the identity and purity of food additives. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of food additives (including flavouring agents) and contaminants, assessments of intake, and the establishment and revision of specifications for food additives. A summary follows of the Committee's evaluations of toxicological and intake data on various specific food additives (alpha-amylase from Bacillus lichenformis containing a genetically engineered alpha-amylase gene from B. licheniformis, annatto extracts, curcumin, diacetyl and fatty acid esters of glycerol, D-tagatose, laccase from Myceliophthora thermophila expressed in Aspergillus oryzae, mixed xylanase, beta-glucanase enzyme preparation produced by a strain of Humicola insolens, neotame, polyvinyl alcohol, quillaia extracts and xylanase from Thermomyces lanuginosus expressed in Fusarium venenatum), flavouring agents, a nutritional source of iron (ferrous glycinate, processed with citric acid), a disinfectant for drinking-water (sodium dichloroisocyanurate) and contaminants (cadmium and methylmercury). Annexed to the report are tables summarizing the Committee's recommendations for ADIs of the food additives, recommendations on the flavouring agents considered, and tolerable intakes of the contaminants considered, changes in the status of specifications and further information requested or desired.

Animals↗

Ochratoxin A from a toxicological perspective.

Ochratoxin A (OTA) is a widespread mycotoxin which is produced mainly by the mould fungi Aspergillus ochraceus and Penicillum verrucosum during the storage of cereals, cereal products and other plant-derived products such as herbs, spices, grapes, etc. By carry over from mouldy fodder, ochratoxin A is also found in pork meat, offal and sausages containing pork blood. When ingested as a food contaminant, OTA is very persistent in human beings with a blood half-life of 35 days after a single oral dosage due to unfavourable elimination toxicokinetics. This renders the toxin among the most frequent mycotoxin contaminants in human blood in the EU, the US, Canada, and elsewhere, where it has been investigated. OTA is neither stored nor deposited in the body, but heterogeneous body distribution may impose serious damage to the kidneys. The toxin was classified a 2B cancer compound, being possibly carcinogenic for humans. It was among the strongest carcinogenic compounds in rats and mice. As the toxicological profile also includes teratogenesis, nephrotoxicity, and immunotoxicity, legislation authorities are currently discussing maximal residue levels (MRL) for OTA in various foodstuffs. In the present article arguments are presented which suggest an acceptable daily intake (ADI) of 1.5 ng OTA/kg body weight and a much lower MRL than 5 microgram OTA/kg cereals and cereal products as has been postulated by the EU commission.

Animals↗

A review of sampling plans and collaboratively studied methods of analysis for aflatoxins.

Aflatoxins are the only food contaminants being monitored routinely on an international scale with methods operating at the order of magnitude of 10 mug/kg. At this concentration level, methods of analysis which can achieve coefficients of variation of 30-40% with recoveries of 70% or greater in interlaboratory collaborative studies can be considered eligible for referee status. In most cases, sample reproducibility is the variable limiting the reliability of methods of analysis. The inherent uncertainty of the identity of chromatographically separated entities requires the application of confirmatory tests to verify that the characteristic measured results from the presence of aflatoxin. The methods are also inoperable without a verification of the identity, purity, and concentration of the reference standards used. Screening methods which reliably eliminate negative samples from further consideration are indispensible for the practical operation of monitoring programs.

Aflatoxins↗

Eight cases of fascioliasis: clinical and microbiological features.

UNLABELLED: Fascioliasis is a parasitic infection caused by the ingestion of food contaminated with Fasciola hepatica. OBJECTIVE AND DESIGN: We report an epidemic of eight cases of fascioliasis in southern France, and describe the clinical features, and the diagnostic and therapeutic tools. RESULTS: Our series includes almost every clinical form described, apart from the pseudotumoral form. Early diagnosis relied mainly on serum assays. Among these, counter-electrophoresis was the first to be positive and therefore appears to be the most useful test, including cases where patients are asymptomatic or mildly ill. Patients were treated with the usual dose of triclabendazole, but six of them required a double dose to recover. CONCLUSION: The use of 20 mg/kg triclabendazole is safe and efficient for patients with acute fascioliasis when a single dose has failed to cure them.

Adult↗

Increasing serum levels of non-DDT-derivative organochlorine pesticides in the younger population of the Canary Islands (Spain).

Organochlorine pesticides are a lipophilic class of chemicals that persist in the environment and tend to accumulate in human tissues for years. They came into widespread use in the late 1940s. Because of their capacity to bioaccumulate and biomagnify in food chains and their toxic effects, most of them were banned in industrialized countries, among them Spain, in the late 1970s and 1980s. In 1998 organochlorine pesticides were determined in a representative sample of a Spanish population (around 690 serum samples from people 6 to 75years old from the Canary Islands). Serum levels of lindane aldrin, dieldrin and endrin, were determined. Our results showed that a high percentage of samples presented detectable levels of some of the organochlorines measured, endrin being the most frequently detected (72%) and at highest concentration (mean 136.7ng/g fat). Mean concentrations of the main cyclodiene evaluated, dieldrin, was lower to those found in other Western populations. However, serum levels of lindane were higher than those described in North European populations. Influence of geographical and sociodemographic factors was evaluated. Urban populations showed the highest levels of dieldrin, while non-urban population showed the highest serum values of lindane, aldrin and endrin. Unexpectedly, serum values of lindane, aldrin and dieldrin were higher in younger than in older people. Subjects under 18years showed almost twice as high serum levels of lindane, aldrin and dieldrin than subjects of 65-75years. These results may well suggest that people living in the Canary Islands have been and are currently exposed to non-DDT-organochlorine pesticides. The type and source of exposure could vary between islands and type of habitat. Contaminated food and/or the environment could be related with this situation.

Adolescent↗

Microcystins (cyanobacteria hepatotoxins) bioaccumulation in fish and crustaceans from Sepetiba Bay (Brasil, RJ).

Blooms of cyanobacteria in water bodies cause serious environmental problems and the occurrence of toxic strains are also related with the human health. Aquatic animals could bioaccumulate microcystins (cyanobacteria hepatotoxins) and so, beyond water, the ingestion of contaminated food represents a human health risk. Recently, WHO recommended a maximum concentration of microcystins (MCYSTs) in drinking water and established the tolerable daily intake (TDI) for consumption of cyanobacteria products contends MCYSTs (0.04 microg(-1) kg(-1) day(-1)). Sepetiba Bay is located in the municipal districts of Rio de Janeiro, Mangaratiba and Itaguai; being an important place of fishing activity. Due to the industrial development in the area, this bay is submitted to different environmental impacts, increasing the organic and industrial pollution. A strain of the nanoplanktonic cyanobacteria Synechocystis aquatilis f. aquatilis that produce MCYSTs was already isolated. In this study, we verified MCYSTs presence in muscle tissue of fish and crustaceans, which were harvested monthly in Sepetiba Bay during 11 months, in order to evaluate the potential risk of their ingestion. MCYSTs were analyzed by immunoassay techniques using the ELISA Microcystin Plate Kit (ENVIROLOGIX INC) and the concentration were expressed as microcystin-LR equivalent. The analyses of seston samples, water, muscle tissues showed the presence of this cyanotoxin in all samples and it was verified that 19% of the animals' samples were above the limit recommended by WHO for human consumption. The maximum value found was of 103.3 microg kg(-1) (TDI 0.52 microg kg(-1) day(-1)) and the minimum, was 0.25 microg kg(-1) in crabs muscle tissue (TDI of 0.001 microg kg(-1) day(-1)). Such data demonstrate that, although in low concentrations, there is already a contamination of fish and crustaceans from Sepetiba Bay. We highlight that the recommended limit refers to healthy adult.

Animals↗

Detection and characterization of norovirus outbreaks in Germany: application of a one-tube RT-PCR using a fluorogenic real-time detection system.

Outbreaks of gastroenteritis caused by Norovirus are an increasing public health problem worldwide. The virus is transmitted by contaminated food and by person-to-person. In Germany, a new health reporting system including Norovirus infections has been introduced in 2001. Norovirus outbreaks (73%) diagnosed in our lab occurred in the first half of the year. Phylogenetic analysis shows that 90% belonged to genogroup II (GGII) with the majority related to prototypes Lordsdale/93 and Tarragona/2001. To date, PCR is the most sensitive and specific method for the detection but several procedures are needed to evaluate the amplification product. To minimize the risk of product carryover, a fast and simple procedure with a minimal number of steps are required. A one-tube RT-PCR method is described using the real-time TaqMan-PCR system. Primer sets and probes for both genogroup I (GGI) and genogroup II (GGII) sequences were developed. The specificity and sensitivity of this method was evaluated on 70 Norovirus positive stool samples of 70 outbreaks in Germany and 34 European samples by comparing the detection rate to those of an in-house RT-nested PCR. Overall, 93% of the PCR positive samples have been detected by the TaqMan-PCR including isolates of four different GGI and seven different GGII genotypes. Using plasmid standards for quantitation, virus loads between 10(2) and 10(10) genomic equivalents/ml stool suspension were found. The advantages of the one-step RT/TaqMan-PCR system for detection and quantitation of Norovirus are the high throughput of clinical samples and a decrease of the risk of contamination.

Caliciviridae Infections↗

Organochlorines, lead, and mercury in Akwesasne Mohawk youth.

Most humans have detectable body burdens of polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB), and p,p'-dichlorophenyldichloroethylene p,p'-DDE), a metabolite of p,p'-dichlorodiphenyltrichloroethane (DDT). Native American communities may be at increased risk of exposure through subsistence-based diets and greater physical contact with contaminated soil and water. In this article we describe the levels of toxicants (PCBs, p,p'-DDE, HCB, mirex, lead, and mercury) among youth 10-17 years old (n = 271) of the Akwesasne Mohawk Nation. Ultratrace, congener-specific PCB analysis of human serum quantitated 83 PCB congeners (plus 18 as pairs/triplets), in addition to p,p'-DDE, HCB, and mirex, and included all major Aroclor-derived congeners typically present in human samples. Twenty congeners (in 16 chromatographic peaks) were detected in 50% or more of the individuals sampled [geometric mean (GM) of the sum of these congeners = 0.66 ppb]. Thirteen congeners (in 10 peaks) were detected in 75% or more of the samples (GM = 0.51 ppb). Of the 20 congeners detected in 50% or more of the samples, 17 had five or more chlorine substitutions. International Union for Pure and Applied Chemistry congeners 118, 101(+90), and 153 were detected in nearly all participants (GM = 0.06 ppb, 0.05 ppb, 0.09 ppb, respectively). p,p'-DDE and HCB were detected in 100% and 98% of the samples (GM: p,p'-DDE = 0.37 ppb; HCB = 0.03 ppb). Mirex was detected in approximately 46% of the samples (GM = 0.02 ppb). No cases of elevated lead level were observed. One participant had a mercury level marginally higher than the U.S. Environmental Protection Agency's current level of concern (0.50 microg/dL). Although differences in analytic methods and participant ages limit comparability, toxicant levels from the Mohawk youth are lower than those associated with severe food contamination (Yusho and Yu-cheng) but similar to other chronically exposed groups.

Adolescent↗

Oral administration of yessotoxin stabilizes E-cadherin in mouse colon.

YTX has been shown to disrupt the E-cadherin-catenin system in cultured epithelial cells, raising some concern that ingestion of seafood contaminated by YTX might favour tumour spreading and metastasis formation in vivo. In order to probe whether YTX might affect cadherin systems in vivo, we have set up a study involving repeated oral dosing of the toxin in mice (1mg/kg/day, for 7 days) and analysis of E-cadherin and N-cadherin in tissue extracts obtained at the end of the dosing scheme, as well as 1 and 3 months after YTX administration. We found that the E-cadherin pools obtained from lung and kidney were not altered by YTX in any of our experimental conditions. Extracts from mouse colon contained intact E-cadherin and an E-cadherin fragment of about 90 kDa (ECRA(90)), displaying a molecular alteration resembling that caused by YTX in cultured cells. We found that the relative proportion of ECRA(90), as compared to intact E-cadherin, was higher in colon extracts from control mice than from YTX-treated animals, indicating that oral administration of YTX to mice stabilizes E-cadherin of mouse colon. No significant difference could be detected in samples prepared from colons obtained 30 or 90 days after termination of YTX treatment. Oral administration of YTX to mice did not lead to a significant increase in the fragments of E-cadherin detectable in serum, neither it altered the N-cadherin pool of mouse heart. Electron microscopy analysis showed no substantial ultrastructural differences between controls and YTX-treated mice. Our findings show that ingestion of food contaminated by YTX poses a low risk of disruption of the E-cadherin system in vivo.

Administration, Oral↗

Sensitive detection of viable Listeria monocytogenes by reverse transcription-PCR.

Detection of pathogens in contaminated food products by PCR can result in false-positive data due to the amplification of DNA from nonviable cells. A new method based on reverse transcription-PCR (RT-PCR) amplification of mRNA for the specific detection of viable Listeria monocytogenes was developed. The expression of three L. monocytogenes genes, iap, hly, and prfA, was examined to determine a suitable target for amplification of RT-PCR. Total RNA from L. monocytogenes was isolated, and following DNase treatment, the RNA was amplified by both RT-PCR and PCR with primers specific for the three genes. Amplicon detection was accomplished by Southern hybridization to digoxigenin-labeled gene probes. The levels of expression of these three genes differed markedly, and the results indicated that the iap gene would provide a good target for development of a specific method for detection of viable L. monocytogenes based on RT-PCR amplification. After a 1-h enrichment, the 371-bp iap-specific product was detected with a sensitivity of ca. 10 to 15 CFU/ml from pure culture. Detection of the 713-bp hly-specific amplicon was ca. 4,000 times less sensitive after 1 h, whereas detection of the 508-bp prfA product showed the lowest level of sensitivity, with detection not observed until after a 5-h enrichment period. The amplification of the iap mRNA was specific for L. monocytogenes. Overall, the assay could be completed in ca. 54 h. The use of RT-PCR amplification for the detection of viable L. monocytogenes was validated in artificially contaminated cooked ground beef. Following a 2-h enrichment incubation, the iap-specific amplification product could be detected in a cooked meat sample that was originally inoculated with ca. 3 CFU/g. These results support the usefulness of RT-PCR amplification of mRNA as a sensitive method for the specific detection of viable L. monocytogenes and indicate that this method may prove useful in the detection of this pathogen in ready-to-eat, refrigerated meat products.

Bacterial Proteins↗