The lack of data on consumer refrigeration temperatures and storage times limits our ability to assess and manage risks associated with microbial hazards. This study addressed these limitations by collecting data on temperatures and storage handling practices of chilled foods. Consumers from 102 households in Uppsala, Sweden, were instructed to purchase seven food items (minced meat, fresh herring fillets, soft cheese, milk, sliced cooked ham, vacuum-packed smoked salmon, and ready-to-eat salad) and to store them using their normal practices. They were interviewed the next day, and food temperatures were measured. In general, there were no significant relations between temperature and characteristics of the respondents (e.g., sex, age, education, age of the refrigerator). Mean storage temperatures ranged from 6.2 degrees C for minced meat to 7.4 degrees C for ready-to-eat salad. Maximum temperatures ranged from 11.3 to 18.2 degrees C. Data were not significantly different from a normal distribution, except for ready-to-eat salad, although distributions other than the normal fitted data better in most cases. Five percent to 20% of the food items were stored at temperatures above 10 degrees C. Most respondents knew the recommended maximum temperature, but less than one fourth claimed to know the temperature in their own refrigerator. Practical considerations usually determined where food was stored. For products with a long shelf life, stated storage times were different for opened and unopened packages. The current situation might be improved if consumers could be persuaded to use a thermometer to keep track of refrigerator temperature.
Wheat is often infected by Fusarium species producing mycotoxins, which may pose health risks to humans and animals. Deoxynivalenol (DON) is the most important Fusarium toxin in Swedish wheat and has previously been shown to be produced mainly by Fusarium graminearum. However, less is known about the co-occurrence of DON and F. graminearum with other toxins and Fusarium species in Sweden. This study examined the distribution of the most important toxigenic Fusarium species and their toxins in winter wheat (2009 and 2011) and spring wheat (2010 and 2011). DNA from seven species was quantified with qPCR and the toxin levels were quantified with a multitoxin analysis method based on liquid chromatography/electrospray ionisation-tandem mass spectrometry (HPLC/ESI-MS/MS). The method enabled detection of many fungal metabolites, including DON, zearalenone (ZEA), nivalenol (NIV), T-2 toxin, HT-2 toxins, moniliformin (MON), beauvericin (BEA), and enniatins (ENNs). It was found that Fusarium poae and Fusarium avenaceum were present in almost all samples. Other common Fusarium species were F. graminearum and F. culmorum, present in more than 70% of samples. Several species occurred at lower DNA levels in 2011 than in other years, but the reverse was true for F. graminearum and Fusarium langsethiae. The most prevalent toxins were ENNs, present in 100% of samples. DON was also common, especially in spring wheat, whereas ZEA and NIV were common in 2009 and in winter wheat, but less common in 2011 and in spring wheat. Only three samples of spring wheat contained T-2 or HT-2 above LOQ. Annual mean levels of several mycotoxins were significantly lower in 2011 than in other years, but the reverse applied for DON. The strongest correlations between mycotoxin and Fusarium DNA levels were found between F. avenaceum and ENNs (r(2) = 0.67) and MON (r(2) = 0.62), and F. graminearum and DON (r(2) = 0.74). These results show that several Fusarium species and toxins co-occur in wheat. The highest toxin levels were detected in spring wheat and DON and ENNs, the latter belonging to the group of so called "emerging toxins", which were the most prevalent toxins and those occurring at the highest levels.
The current methods for the detection of pathogenic Yersinia enterocolitica bacteria in food are time consuming and inefficient. Therefore, we have developed and evaluated in-house a TaqMan probe-based real-time PCR method for the detection of this pathogen. The complete method comprises overnight enrichment, DNA extraction, and real-time PCR amplification. Also included in the method is an internal amplification control. The selected primer-probe set was designed to use a 163-bp amplicon from the chromosomally located gene ail (attachment and invasion locus). The selectivity of the PCR method was tested with a diverse range (n ؍ 152) of related and unrelated strains, and no false-negative or false-positive PCR results were obtained. The sensitivity of the PCR amplification was 85 fg purified genomic DNA, equivalent to 10 cells per PCR tube. Following the enrichment of 10 g of various food samples (milk, minced beef, cold-smoked sausage, fish, and carrots), the sensitivity ranged from 0.5 to 55 CFU Y. enterocolitica. Good precision, robustness, and efficiency of the PCR amplification were also established. In addition, the method was tested on naturally contaminated food; in all, 18 out of 125 samples were positive for the ail gene. Since no conventional culture method could be used as a reference method, the PCR products amplified from these samples were positively verified by using conventional PCR and sequencing of the amplicons. A rapid and specific real-time PCR method for the detection of pathogenic Y. enterocolitica bacteria in food, as presented here, provides a superior alternative to the currently available detection methods and makes it possible to identify the foods at risk for Y. enterocolitica contamination.
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