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Rotavirus survival and stability in foods as determined by an optimised plaque assay procedure.

Tissue culture adapted rotavirus strains were propagated in MA104 and CaCo2 cells using standard cell culture procedures. The progress of infection was monitored by examining for a cytopathic effect, and for the presence of viral RNA in the tissue culture supernatant as determined by a guanidinium-based method. Subsequently, an effective plaque assay for rotavirus was developed using MA104 cells by optimising the adsorption time (2 h) and the levels of fetal calf serum (2.5%) in the overlay medium. Tragacanth gum was used in the overlay medium to immobilize the virus, and plaques were subsequently stained with 1% crystal violet. Using this optimised plaque assay, the survival of rotavirus following exposure to heat and UV irradiation was evaluated by enumerating the clear plaques. It was shown that 60 degrees C for 10 min was sufficient to reduce the viral titer by at least 7 logs, and 50 mJ of UV irradiation was sufficient to reduce the initial viral titer by > 2.5 logs. This optimised plaque assay was also used to determine the survival and stability of rotavirus from a range of experimentally contaminated foods including fruit juice, formula milk and lettuce.

Cell Line↗

Absorption, accumulation and biological effects of depleted uranium in Peyer's patches of rats.

The digestive tract is the entry route for radionuclides following the ingestion of contaminated food and/or water wells. It was recently characterized that the small intestine was the main area of uranium absorption throughout the gastrointestinal tract. This study was designed to determine the role played by the Peyer's patches in the intestinal absorption of uranium, as well as the possible accumulation of this radionuclide in lymphoid follicles and the toxicological or pathological consequences on the Peyer's patch function subsequent to the passage and/or accumulation of uranium. Results of experiments performed in Ussing chambers indicate that the apparent permeability to uranium in the intestine was higher (10-fold) in the mucosa than in Peyer's patches ((6.21+/-1.21 to 0.55+/-0.35)x10(-6)cm/s, respectively), demonstrating that the small intestinal epithelium was the preferential pathway for the transmucosal passage of uranium. A quantitative analysis of uranium by ICP-MS following chronic contamination with depleted uranium during 3 or 9 months showed a preferential accumulation of uranium in Peyer's patches (1355% and 1266%, respectively, at 3 and 9 months) as compared with epithelium (890% and 747%, respectively, at 3 and 9 months). Uranium was also detected in the mesenteric lymph nodes ( approximately 5-fold after contamination with DU). The biological effects of this accumulation of depleted uranium after chronic contamination were investigated in Peyer's patches. There was no induction of the apoptosis pathway after chronic DU contamination in Peyer's patches. The results indicate no change in the cytokine expression (Il-10, TGF-beta, IFN-gamma, TNF-alpha, MCP-1) in Peyer's patches and in mesenteric lymph nodes, and no modification in the uptake of yeast cells by Peyer's patches. In conclusion, this study shows that the Peyer's patches were a site of retention for uranium following the chronic ingestion of this radionuclide, without any biological consequences of such accumulation on Peyer's patch functions.

Animals↗

The protective effect of alcohol on the occurrence of epidemic oyster-borne hepatitis A.

Limited data indicate that drinking alcoholic beverages along with eating food contaminated with Shigella or Salmonella decreases the risk and/or the severity of illness. No such study has been reported following exposure to a viral pathogen. During an oyster-borne outbreak of hepatitis A, we studied the effect of ingestion of alcoholic beverages concomitant with consumption of contaminated oysters. The analysis was restricted to 51 cases and 33 controls who had consumed the implicated raw oysters. After controlling for potential confounders, we found a protective effect for beverages that have an alcohol concentration of greater than or equal to 10% (odds ratio = 0.1, 95% confidence interval = 0.02-0.9), but not for beverages with an alcohol concentration of less than 10% (odds ratio = 0.7, 95% confidence interval = 0.2-2.9).

Adult↗

Predicting pesticide residues to reduce crop contamination.

The system of dynamics between pesticides and plants is reviewed, and a conceptual model capable of reflecting the necessary qualitative and structural peculiarities is proposed as a means of predicting residue levels. The degradation processes of various chemical classes of pesticides in plants under different conditions of use are analyzed. Formulas are developed that enable recognition and estimation of residue levels and duration of retention for "new" pesticides and "old" substances under varying treatment conditions. Estimated data are verified to provide positive assessments of the accuracy of the predictions. Mathematical modeling as a means of perception is stressed. With this method, the outlet value can be controlled by changing such inlet parameters as application rate, frequency of treatments, types of plants, and so on. Residue levels are predicted for different combinations of use conditions in various climatic-geographical regions. The method of estimation also enables the development of important standards such as post-treatment waiting intervals. A more flexible technique can be employed, in which specific periods are established for different plants under various treatment conditions. Thus, both unjustified shortened waiting intervals or unnecessarily elongated intervals periods can be avoided. The greatest value of the modeling approach is that information can be obtained on the degree of potential food contamination and major standards can be developed without the need for extensive experimental use of pesticides under actual conditions. This technique fully considers optimal use conditions for agrochemicals in terms of human requirements and protection of plants. Mathematical modeling protects the environment while enabling the speedy selection of safe parameters for pesticide use conditions nd realization of significant savings in manpower and time. Overall, this estimation method appears to be an efficient link in the broad system of preventing environmental pesticide contamination and protecting human health.

Humans↗

A small outbreak of listeriosis associated with smoked mussels.

Two perinatal listeriosis cases in Auckland, New Zealand, which were diagnosed during November and December 1992 gave histories of consuming Brand X smoked mussels. Listeria monocytogenes was isolated from an unopened packet of mussels collected from the refrigerator of one of the cases. Cultures of L. monocytogenes from the two patients were indistinguishable from those recovered from the mussels by serogrouping and DNA macrorestriction analysis using pulsed-field gel electrophoresis (PFGE). PFGE analysis of isolates of an additional fifteen clinical cases caused by L. monocytogenes serogroup 1/2 which occurred in New Zealand during 1991 and 1992, revealed two more isolates with indistinguishable PFGE patterns. PFGE analysis of a further 222 L. monocytogenes serogroup 1/2 isolated in New Zealand from food and the environment did not reveal any more cultures with the Brand X PFGE type. A combination of serotyping, phage typing, DNA restriction fragment length polymorphism analysis (RFLP), cadmium and arsenic sensitivity testing, and PFGE analysis was used to subtype 38 isolates of L. monocytogenes. The isolates comprised: (a) the isolates from four patients; (b) 26 isolates from 15 packets of Brand X mussel products from retail sale, the processing factory, and the refrigerator of one of the patients; (c) seven isolates from environmental swabs taken in the processing factory; and (d) an isolate from Brand X mussel product from a wholesaler in the United Kingdom. The isolates from three of the clinical cases, 26 of the products and four from the factory environment were indistinguishable using all the subtyping systems. This is the first description from New Zealand of a link between cases of listeriosis, a contaminated food, and the food production environment which was microbiologically confirmed using a combination of subtyping methods for L. monocytogenes.

Animals↗

[Two epidemic diseases in Blois region in the 19th century].

Lathyrism is an ancient neurological disorder caused in human by food contamination with seeds of Lathyrus species. Based on new documents, this paper describes the exact sanitary actions done in the discrict of Blois in 1829. Official reports due to Dr. Desparanches, M.D., describe clinical cases, causes of the disease and its extension in the population. A bacterial dysentry occured mostly in children and poor families in the summer of 1858 in the same district. An official report due to Dr. Monneau, M.D., describes the extension and possible causes of the diseases. In the two reports, sucessful pharmaceutical treatments and public sanitary measures are described.

Disease Outbreaks↗

Epidemiological study of a food-borne outbreak of enterotoxigenic Escherichia coli O25:NM by pulsed-field gel electrophoresis and randomly amplified polymorphic DNA analysis.

This study investigated the applicability of molecular epidemiological techniques to the identification of the causal agent of an outbreak of diarrhea caused by ingestion of food contaminated with enterotoxigenic Escherichia coli (ETEC). The outbreak occurred at four elementary schools in July 1996 and affected more than 800 people. Illness was most strongly associated with eating tuna paste (relative risk, 1.79; 95% confidence interval = 1.16 to 2.79; P = 0.0001). To evaluate the epidemiological characteristics of the pathogen, the DNAs from numerous isolated ETEC strains were subjected to randomly amplified polymorphic DNA analysis, pulsed-field gel electrophoresis of nuclease S1-treated plasmid DNA, and analysis of genomic DNA restriction fragment length polymorphisms. All ETEC isolates were of the O25:NM (nonmotile) serotype, which carries a heat-stable enterotoxin Ib gene. Genotypic analysis demonstrated that the strains isolated from the patients at all four schools were identical. The isolates of ETEC O25:NM obtained from the tuna paste that had been served for lunch at these schools were genetically indistinguishable from those isolated from the patients. Results suggest that this outbreak was food borne. The molecular biology-based epidemiological techniques used in this study were useful in characterizing the causal agent in this food-borne epidemic.

Animals↗

Molecular typing and virulence markers of Yersinia enterocolitica strains from human, animal and food origins isolated between 1968 and 2000 in Brazil.

Molecular typing and virulence markers were used to evaluate the genetic profiles and virulence potential of 106 Yersinia enterocolitica strains. Of these strains, 71 were bio-serotype 4/O : 3, isolated from human and animal clinical material, and 35 were of biotype 1A or 2 and of diverse serotypes, isolated from food in Brazil between 1968 and 2000. Drug resistance was also investigated. All the strains were resistant to three or more drugs. The isolates showed a virulence-related phenotype in the aesculin, pyrazinamidase and salicin tests, except for the food isolates, only two of which were positive for these tests. For the other phenotypic virulence determinants (autoagglutination, Ca(++) dependence and Congo red absorption), the strains showed a diverse behaviour. The inv, ail and ystA genes were detected in all human and animal strains, while all the food isolates were positive for inv, and 3 % of them positive for ail and ystA. The presence of virF was variable in the three groups of strains. The strains were better discriminated by PFGE than by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). A higher genomic similarity was observed among the 4/O : 3 strains, isolated from human and animal isolates, than among the food strains, with the exception of two food strains possessing the virulence genes and grouped close to the 4/O : 3 strains by ERIC-PCR. Unusually, the results revealed the virulence potential of a bio-serotype 1A/O : 10 strain, suggesting that food contaminated with Y. enterocolitica biotype 1A may cause infection. This also suggests that ERIC-PCR may be used as a tool to reveal clues about the virulence potential of Y. enterocolitica strains. Furthermore, the results also support the hypothesis that animals may act as reservoirs of Y. enterocolitica for human infections in Brazil, an epidemiological aspect that has not been investigated in this country, confirming data from other parts of the world.

Agglutination Tests↗

The home environment and salmonellosis in children.

OBJECTIVE: To explore the role of foods and the home environment in the development of Salmonella infections in infants and children. METHODS: Home investigations were conducted of patients younger than 4 years of age infected with Salmonella. Cultures were obtained from foods, persons residing in the home, animals/pets/insects, and environmental sources. Like serotypes encountered in the index patients and isolates from the home underwent typing with pulsed-field gel electrophoresis. RESULTS: Home inspections were conducted in approximately 66% of eligible homes on the average of 3.4 days after the confirmation of the Salmonella isolate. A total of 526 cultures from 50 homes were obtained from foods (120), household members (73), refrigerators (52), water (47), countertops (46), soil (42), can-openers (36), vacuum cleaners (34), animals/pets/insects (26), and others (50). Isolates with a serotype identical to those in the index patient were found in 16 homes, 3 of which included an isolate of a second serotype, and an isolate of a different serotype was recovered in 3 homes. The pulsed-field gel electrophoresis patterns of the isolates of identical serotypes from the subjects and from their environment were indistinguishable in all but 2 patients. Among isolates of the same serotype encountered in different homes, all patterns were different. The identical serotype was found in multiple locations (4), dirt surrounding front doors (4), household members (3), vacuum cleaner (1), animals/pets/insects (1), and a refrigerator shelf (1). CONCLUSIONS: These data illustrate the importance of the child's environment in the development of salmonellosis. Clinicians should concentrate on educating the parents about the environmental spread of Salmonella. Contaminated foods in the home play a less significant role in the infection of infants and children.

Arkansas↗

Influence of subchronic exposure to low dietary deoxynivalenol, a trichothecene mycotoxin, on intestinal absorption of nutrients in mice.

During a 6-wk feeding trial, effects of low dietary deoxynivalenol (DON; 0, 0.1, 1 and 10 ppm) on food consumption and weight gain were investigated in male mice. Food intake was similar in all four dietary groups. Weight gain in the group receiving 10 ppm DON was significantly (P less than 0.01) reduced. At the end of the feeding period, test animals were sacrificed and absorption of water, D-glucose, L-leucine, L-tryptophan, 5-methyltetrahydrofolic acid and iron was measured in isolated perfused jejunal segments in vitro. No effects were observed on absorption of water, leucine, tryptophan and iron. However, at a dietary DON concentration of 10 ppm, a slightly but significantly (P less than 0.05) reduced transfer of glucose was measured. Furthermore, transfer as well as tissue accumulation of 5-methyltetrahydrofolic acid in the jejunal segment were both significantly decreased up to 50%. Heavy metal and trace element content was determined in liver, kidney and small intestine. Manganese and molybdenum content in liver tissue was reduced with a DON concentration of 10 ppm in the diet. The findings indicate that subchronic ingestion of DON, in concentrations occurring in contaminated food and feed, results in an impairment of intestinal transfer and uptake of nutrients such as glucose and 5-methyltetrahydrofolic acid.

Animals↗

Matrix-assisted laser desorption ionization-time of flight mass spectrometry for the discrimination of food-borne microorganisms.

A methodology based on matrix-assisted laser desorption ionization-time of flight mass spectrometry of intact bacterial cells was used for rapid discrimination of 24 bacterial species, and detailed analyses to identify Escherichia coli O157:H7 were carried out. Highly specific mass spectrometric profiles of pathogenic and nonpathogenic bacteria that are well-known major food contaminants were obtained, uploaded in a specific database, and made available on the Web. In order to standardize the analytical protocol, several experimental, sample preparation, and mass spectrometry parameters that can affect the reproducibility and accuracy of data were evaluated. Our results confirm the conclusion that this strategy is a powerful tool for rapid and accurate identification of bacterial species and that mass spectrometric methodologies could play an essential role in polyphasic approaches to the identification of pathogenic bacteria.

Bacteria↗

[Pneumothorax in distomatosis].

INTRODUCTION: Distomatosis is a parasite infection following ingestion of food contaminated by Fasciola hépatica, the tropism of which is essentially hepatic. The mechanism of this exceptional complication is discussed in two cases of pneumothorax. OBSERVATIONS: Two 35 and 60 year-old patients presenting spontaneous pneumothorax during distomatosis, the first revealing the disease, the second suggesting the evolution of a severe form of the disease. The surgical biopsy in the first patient revealed a granulomatous pleuro-pneumopathy rich in giant cells. DISCUSSION: Pulmonary manifestations of distomatosis are frequent during the invasive phase. Their immunoallergic origin can be questioned and emphasis should be placed on lesions secondary to aberrant parasite migrations.

Adult↗

Antifungal activity of Mexican oregano (Lippia berlandieri Shauer).

Antifungal and sensorial properties of spices have been recognized for years. The antifungal compounds are products of the plant's secondary metabolism, and the action of those compounds could be used to inhibit the growth of spoilage and pathogenic microorganisms in food. Mexican oregano (Lippia berlandieri) grows wildly in the desert zone of Mexico and is usually added to regional foods. The goal of this study was to evaluate the antifungal activity of Mexican oregano versus food-contaminant fungi. Fungi were isolated from spoiled fruit and vegetables and identified according to morphological characteristics. The antifungal activity of oregano was evaluated by radial growth measurement on potato dextrose agar added with dried oregano (0.25 to 4.0%). The essential oil antifungal activity of oregano was also evaluated by the diffusion well test. Twenty-one fungal strains were isolated, which included Penicillium, Geotrichum, Aspergillus, and Bipolaris. In seven of the 21 strains, no inhibitory effect was observed at either concentration of oregano. An increase in growth at the lower or higher concentrations of oregano, when compared to the control, was observed in two fungal strains; in 12 strains, a strong inhibitory effect of oregano was evident. The oregano essential oil was inhibitory to all fungal strains, but there were differences in the extent of the effect. Although the antifungal effect of oregano is strongly established, there was a differential effect with the fungal strains studied. Besides pathogenic fungi and bacteria, microbial spoilage flora should be considered when the addition of spices for food preservation is proposed.

Antifungal Agents↗

Genotoxicity assessment of deoxynivalenol in the Caco-2 cell line model using the Comet assay.

The genotoxic risk associated with deoxynivalenol (DON), a prevalent trichothecene mycotoxin which contaminates cereal-based products has not yet been deeply explored. In this work, the alkaline version of the Comet assay was used to evaluate DNA damage stemming from DON exposure in both dividing and differentiated Caco-2 cells, an epithelial intestinal cell line. To avoid false positive results, cytotoxic and apoptotic thresholds were firstly established using the MTS and neutral red assays and the Hoestch staining method, respectively. Dividing cells were found to be more sensitive to DON than differentiated cells and the lowest IC(10) (0.5 microM) obtained for dividing cells exposed for 72 h was used as the highest working concentration in the genotoxicity study. Both differentiated and dividing cells responded with a dose-dependent relationship to DON in terms of DNA damage in the 0.01-0.5 microM range. These results demonstrated the existence of a genotoxic potential for DON at low concentrations compatible with actual exposure situations and calls for additional studies to determine the functional consequences which could be taken into account for the risk assessment of this food contaminant.

Apoptosis↗

Ovipositional response, developmental effects and toxicity of hexavalent chromium to Megaselia scalaris, a terrestrial detritivore.

The effects of hexavalent chromium (Cr VI) on ovipositional response, development, and survival of a common terrestrial detritivore, Megaselia scalaris (Diptera: Phoridae), were assessed in the laboratory. Ovipositing females did not discriminate between substrates containing 0, 50, 500, or 1000 microg/g, indicating a lack of avoidance behavior. Eggs placed on artificial diets containing up to 1000 microg/g either did not absorb Cr VI or were unaffected as measured by eclosion rates. However, development and survival of larvae were significantly reduced at the higher concentrations tested. Concentrations of 500 microg/g in their food increased larval development times by nearly 65%. At 1000 microg/g, larval developmental times doubled. The time required from onset to completion of pupariation was not significantly different regardless of Cr VI concentration. Although males eclosed before females, there was no significant difference between the sexes in the time required for adult eclosion. In addition, there were no significant differences in the percentage of males and females emerging from any of the treatments. At concentrations of 500 or 1000 microg/g, Cr VI decreased larval survival. Survival was reduced by 44.3% at 500 microg/g as compared with the controls. There was no additional mortality from the onset of the puparial stage to adult eclosion for larvae fed diets containing 500 microg/g Cr VI. At 1000 microg/g Cr VI, larval survival decreased by 86.6%. An additional 7.4% mortality was recorded in the puparial stage, for a decrease in total survival (larval plus puparial stages) of 94%. Thus, nearly all of the observed mortality occurred during the larval stage rather than the puparial stage. The population level implications of lack of avoidance of contaminated food and the effects of increased developmental times and reduced survivorship are discussed.

Animals↗

Methicillin (Oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans.

From May 2001 to April 2003, various types of specimens from cattle, pigs, and chickens were collected and examined for the presence of methicillin (oxacillin)-resistant Staphylococcus aureus (MRSA). S. aureus was isolated and positively identified by using Gram staining, colony morphology, tests for coagulase and urease activities, and an API Staph Ident system. Among 1,913 specimens collected from the animals, 421 contained S. aureus; of these, 28 contained S. aureus resistant to concentrations of oxacillin higher than 2 micro g/ml. Isolates from 15 of the 28 specimens were positive by PCR for the mecA gene. Of the 15 mecA-positive MRSA isolates, 12 were from dairy cows and 3 were from chickens. Antimicrobial susceptibility tests of mecA-positive MRSA strains were performed by the disk diffusion method. All isolates were resistant to members of the penicillin family, such as ampicillin, oxacillin, and penicillin. All isolates were also susceptible to amikacin, vancomycin, and trimethoprim-sulfamethoxazole. To determine molecular epidemiological relatedness of these 15 animal MRSA isolates to isolates from humans, random amplified polymorphic DNA (RAPD) patterns were generated by arbitrarily primed PCR. The RAPD patterns of six of the isolates from animals were identical to the patterns of certain isolates from humans. The antibiotypes of the six animal isolates revealed types similar to those of the human isolates. These data suggested that the genomes of the six animal MRSA isolates were very closely related to those of some human MRSA isolates and were a possible source of human infections caused by consuming contaminated food products made from these animals.

Animals↗

Uptake and elimination of benzo[a]pyrene in the terrestrial isopod Porcellio scaber.

In isopods from contaminated sites relatively low levels of high molecular weight polycyclic aromatic hydrocarbons (PAHs) have been observed, which may be caused by either a low bioavailability or a high elimination rate. To shed light on this, the uptake and elimination rates of benzo[a]pyrene were estimated for the isopod Porcellio scaber. The isopod was fed contaminated food (100 microg benzo[a]pyrene/g dwt) for seven weeks followed by four weeks of untreated food. The hindguts of the animals were removed prior to analysis to exclude food material from the body. Benzo[a]pyrene concentrations were log-normally distributed among the individuals. The high inter-individual variation in benzo[a]pyrene content could not be explained from differences in sex, the estimated amount of food in the hindgut, or the body weight. A one-compartment model fitted to the isopod concentrations estimated an assimilation rate of 2.9 microg benzo[a]pyrene/g dwt day, an elimination rate constant of 1.1/day and an equilibrium concentration of 2.5 microg benzo[a]pyrene/g dwt. According to the model 68% of the isopod population had an equilibrium concentration between 1.0 and 7.2 microg benzo[a]pyrene/g dwt day with a benzo[a]pyrene half-life ranging between 0.4 and 1.3 days. The assimilation efficiency was estimated at 20 to 40% of the ingested benzo[a]pyrene. The tissue distribution of benzo[a]pyrene was investigated in a separate experiment. Trace levels of benzo[a]pyrene were detected in haemolymph samples, demonstrating absorption and transport of the compound in the isopod. It is concluded that dietary benzo[a]pyrene is available for uptake to the isopod and that low residues of the compound observed in field isopods are the result of a high elimination rate rather than a reduced bioavailability. As PAHs from soil appear to be available to soil invertebrates, the widespread contamination of the soil from atmospheric emissions is of some concern, especially since the observed elimination may be linked to metabolic activation of PAHs.

Animals↗

Traveler's diarrhea: epidemiology and impact on visitors to Fortaleza, Brazil.

OBJECTIVE: To assess the epidemiology and impact of traveler's diarrhea (TD) among visitors to the city of Fortaleza, Ceará, Brazil, as part of a global study on TD carried out in four countries. METHODS: Within a cross-sectional survey, questionnaires were completed by departing travelers at the Fortaleza airport between March 1997 and February 1998. The questions inquired about demographics, duration of stay, reason for their visit, pretravel health advice they had received, risky food and beverage consumption while in Fortaleza, and quality of life during the visit to Fortaleza in relation to having or not having contracted TD. RESULTS: A total of 12,499 questionnaires were analyzed. The most common reason that the visitors gave for their travel to Fortaleza was a holiday (60.3%). The total diarrhea attack rate was 13.4%. Younger people (< 36 years) had significantly higher TD attack rates than did older persons. Using a logistic regression model, we investigated the visitors' risk factors, including age, gender, length of stay, and trip's purpose. According to that analysis, characteristics that are slightly predictive of TD are gender, length of stay, and visiting as a tourist rather than for some other purpose. Characteristics that protect against contracting TD include being older and traveling for business rather than for some other reason. Of those who were incapacitated by TD, the mean duration of the impairment was 42 hours. CONCLUSIONS: TD affected the travel plans and activities of many of the visitors to Fortaleza. Further, although aware of the health risks, the majority of those travelers did not avoid all potentially contaminated food or beverage items. Given this pattern of behavior, future efforts to combat TD may have to depend on such other alternative strategies as new vaccines.

Adolescent↗