Mats Lindblad

Home Storage Temperatures and Consumer Handling of Refrigerated Foods in Sweden

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.

Deoxynivalenol and other selected Fusarium toxins in Swedish wheat--occurrence and correlation to specific Fusarium species.

Fusarium moulds frequently contaminate oats and other cereals world-wide, including those grown in Northern Europe. To investigate the presence of toxigenic Fusarium species and their toxins in oats, samples were taken during 2010 and 2011 in three geographical regions of Sweden (east, west, south). The samples were analysed by real-time PCR for the specific infection level of seven Fusarium species associated with oats and other cereals (Fusarium poae, Fusarium graminearum, Fusarium langsethiae, Fusarium culmorum, Fusarium tricinctum, Fusarium sporotrichioides and Fusarium avenaceum) and with a multi-mycotoxin method based on liquid chromatography/electrospray ionisation-tandem mass spectrometry (HPLC/ESI-MS/MS) for the detection of many fungal metabolites, including deoxynivalenol (DON), zearalenone (ZEA), nivalenol (NIV), T-2 toxin, HT-2 toxins, moniliformin (MON), beauvericin (BEA) and enniatins (ENNs). Most samples contained at least four of the seven Fusarium species analysed and F. poae, F. langsethiae and F. avenaceum were present in approximately 90-100% of all samples. The most common toxins detected were DON, NIV, BEA and ENNs, which were present in more than 90% of samples. Most Fusarium species and their toxins occurred in higher concentrations in 2010 than in 2011, with the exception of DON and its main producer F. graminearum. Significant regional differences were detected for some moulds and mycotoxins, with higher levels of F. graminearum, DON and ZEA in western Sweden than in the east (P<0.05) and higher levels of F. tricinctum and MON in the south (P<0.05). Correlation analysis showed significant correlations between many Fusarium species and toxin levels. For example, F. tricinctum was significantly correlated to F. avenaceum (r = 0.72, P<0.001), DON to ZEA (r = 0.52, P<0.001), DON to F. graminearum (r = 0.77, P<0.001) and the sum of T-2 and HT-2 to F. langsethiae (r = 0.77, P<0.001). The multi-toxin approach employed allowed simultaneous detection of many Fusarium mycotoxins in each sample. In combination with real-time PCR analysis of seven toxigenic Fusarium spp., the results gave an overall picture of the presence of Fusarium and their toxins in Swedish oats and revealed significant annual and regional differences. This is the first study of the so-called emerging mycotoxins (e.g., ENNs, MON and BEA) in oats grown in Sweden.

Aspergillus nomius, an important aflatoxin producer in Brazil nuts?

The relationship between aflatoxin B1 and G1 was examined in samples from 199 aflatoxin contaminated lots of inshell Brazil nuts imported to Europe. In most of the samples, the relationship between B1 and G1 were approximately 50/50 indicating that the major responsible aflatoxin producing fungi cannot be Aspergillus flavus, which produces solely B aflatoxins. Fungal strains were isolated from two batches of Brazil nuts and isolates of both A. nomius and A. flavus could be identified. The A. nomius isolates were good producers of both B and G aflatoxins, while the A. flavus strains only produced B aflatoxins. In conclusion, this study suggests that A. nomius is an important producer of aflatoxins in Brazil nuts and that its occurrence, and possibly other B and G aflatoxin producers, should be further examined since this may influence strategies for prevention and control of aflatoxins in Brazil nuts.

Predicting noncompliant levels of ochratoxin A in cereal grain from Penicillium verrucosum counts

Aims:  To model the probability of exceeding the European legislative limit of 5 μg ochratoxin A (OTA) per kilogram grain in relation to Penicillium verrucosum levels and storage conditions, and to evaluate the possibilities of using P. verrucosum colony counts for predicting noncompliant OTA levels.

Inactivation of Escherichia coli, Listeria monocytogenes and Yersinia enterocolitica in fermented sausages during maturation/storage

The purpose of this study was to evaluate maturation and storage conditions as a way to increase the safety of non-heat treated fermented sausages. The specific objectives were to investigate the effects of storage time and temperature on the levels of Escherichia coli, Listeria monocytogenes and Yersinia enterocolitica in fermented sausages and in broth, and to validate how well the broth experiments and some published models can predict inactivation in sausage. One strain each of E. coli, L. monocytogenes and Y. enterocolitica with induced acid tolerance was inoculated into sausage batters representing a typical Swedish recipe for cold-smoked sausages. The sausages were fermented at 27 degrees C for 39 or 48 h and then stored at different temperatures (8, 15, or 20-22 degrees C) for up to 44 days. The levels of the experimental strains, lactic acid bacteria, and pH, a(w), and lactic acid was measured during the maturation/storage period. Inactivation in BHI broths adjusted to pH 4.4 or 4.6, water activity of 0.93, and with 1, 1.3 or 2% lactic acid added was also studied. For all strains inactivation rates increased with temperature in both broths and sausages. At 8 degrees C the storage time required for a one-log reduction in sausage ranged from 21 days for E. coli, >16 days for L. monocytogenes, to 18 days for Y. enterocolitica. At temperatures of 20 degrees C or more, the storage time needed for a one log reduction was shorter: between 7 to 11 days for E. coli, 4 to 7 days for L. monocytogenes, and 1 to 4 days for Y. enterocolitica. A published model based on temperature only yielded a good prediction of E. coli inactivation in sausage. A linear model based on the rate estimated in broth yielded a fair prediction of L. monocytogenes inactivation. The performance of other inactivation models validated was unsatisfactory. Significant E. coli growth which occurred in batters without salt during initial phases of fermentation resulted in a subsequent increased inactivation rate, possibly due to increased susceptibility to stress of exponential phase bacteria. The results indicate that the practice of utilising a short maturation period and storage at refrigeration temperatures may result in unsatisfactory reductions of pathogens if present. Thus, inclusion of a maturation period above refrigeration temperatures before distribution may increase the safety of these products.

Escherichia coli with extended-spectrum beta-lactamases or transferable AmpC beta-lactamases and Salmonella on meat imported into Sweden

The presence of Enterobacteriaceae producing extended spectrum beta-lactamases (ESBL) or transferable AmpC beta-lactamases (pAmpC) is increasingly being reported in humans and animals world-wide. Their occurrence in food-producing animals suggests that meat is a possible link between the two populations. This study investigated the occurrence and characteristics of Salmonella and ESBL- or pAmpC-producing E. coli in 430 samples of beef, pork and broiler meat imported into Sweden, in order to provide data required for assessing the potential public health risk of these bacteria in food. Depending on region of origin, ESBL/pAmpC-producing E. coli were found in 0-8% of beef samples, 2-13% of pork samples and 15-95% of broiler meat samples. The highest prevalence was in South American broiler meat (95%), followed by broiler meat from Europe (excluding Denmark) (61%) and from Denmark (15%). Isolates from meat outside Scandinavia were generally defined as multiresistant. A majority of the ESBL/pAmpC genes were transferable by conjugation. Bla(CTX-M-2) and bla(CTX-M-8) were the dominant genes in E. coli from South American broiler meat, whereas bla(CMY-2) and bla(CTX-M-1) dominated in European meat. The majority of bla(CMY-2) and bla(CTX-M-1) were situated on plasmids of replicon type incK and incI1, respectively. The same combinations of ESBL/pAmpC genes and plasmids have been described previously in clinical human isolates. Salmonella was found in five samples tested, from European pork and broiler meat. No Salmonella isolate was resistant to third-generation cephalosporins. In conclusion, meat imported into Sweden, broiler meat in particular, is a potential source of human exposure to ESBL- and pAmpC-producing E. coli.

Growth of aflatoxigenic moulds and aflatoxin formation in Brazil nuts

The present study aimed at gaining more knowledge of the growth of aflatoxigenic moulds and aflatoxin production in Brazil nuts in relation to humidity conditions and storage time. For this purpose, the growth of aflatoxigenic moulds and the increase in aflatoxin levels in Brazil nuts was studied in the laboratory at temperature and humidity conditions that are relevant for the Amazon region. Fresh unprocessed Brazil nuts in shell were inoculated with an aflatoxin producing strain of Aspergillus nomius previously isolated from Brazil nuts. The nuts were stored at 27 °C in combination with 97, 90 or 80% surrounding relative humidity in a respirometer for up to 3 months. The General Linear Model (GLM) was used for evaluation of the effect of water activity and time on aflatoxigenic mould levels and on aflatoxin levels, as well as the relationship between mould and aflatoxin levels. During storage at the highest relative humidity (97%) aflatoxin formation occurred rapidly, whereas storage at 90% relative hum...

Consumers' ability to discriminate aflatoxin-contaminated Brazil nuts

The objectives of the study were to investigate the extent to which consumers can separate nuts with a high content of aflatoxin from sound nuts, and whether sorting results can be improved by information or whether they are affected by certain factors. A test panel consisting of 100 subjects was asked to crack 300 g Brazil nuts and to sort the nuts into those they considered edible and inedible. The test showed that consumers can, on current behaviour, discriminate aflatoxin-contaminated Brazil nuts to a significant extent. The median and the 95th percentile of the total concentrations of aflatoxins (B1, B2, G1, G2) in the samples before sorting were 1.4 and 557 µg kg−1, respectively, and in the edible fractions after sorting 0.4 and 56 µg kg−1, respectively. Given that levels of aflatoxins before sorting exceed either 2 µg aflatoxin B1 kg−1 or totally 4 µg aflatoxins kg−1, there was no effect of aflatoxin concentrations before sorting on the probability of exceeding these thresholds in the edible fraction. This means that similar sorting results were obtained for samples with aflatoxin levels exceeding either of the two thresholds, irrespective of if the thresholds were exceeded with a few µg kg−1 or up to more than 1000 µg kg−1. None of the tested factors (such as sex, age, level of education, ethnic background or knowledge of mycotoxins) had any effects on the probability of exceeding either of the two aflatoxin thresholds.

The effect of undissociated lactic acid on Staphylococcus aureus growth and enterotoxin A production.

The potential of Staphylococcus aureus cheese isolates to grow and produce staphylococcal enterotoxin A under conditions typical for cheese making was investigated in three broth experiments. The effect of the concentration of undissociated lactic acid (HLac) in conjunction with specific pH values was studied by adjusting pH at a single concentration of lactic acid. First, the time-to-growth of S. aureus was modelled by using survival analysis and absorbance data obtained from an automated turbidity reader. The fitted model describes the time to growth and indicates the growth ⁄ no growth boundary of S. aureus as a function of HLac concentration, temperature and water activity. Second, growth rates and lag times of S. aureus were estimated after two different pre-treatments in skim milk at three HLac concentrations and two temperatures based on optical detection times of serial dilutions of bacterial solutions. Growth rates differed between strains, and increased with increasing temperature and decreasing HLac concentration. Preliminary results indicate that lag times were dependent on pre-treatment suggesting that the growth potential of S. aureus in cheese curd may be greater if milk is used immediately after milking compared to holding at 4°C after milking. Third, growth, inactivation, and enterotoxin A production of S. aureus strains were investigated at twelve combinations of HLac concentration and temperature. Concentrations of enterotoxin A increased linearly during the first four days, with a production rate increasing with increasing temperature and decreasing HLac concentration. Significant amounts of enterotoxin A were produced during extended incubation, up to 14days, but then initial pH had changed. This highlights a potential limitation of modelling based on the initial environmental conditions in batch experiments. In summary, ranges of time-to-growth, growth rates, lag times and enterotoxin A production rates of S. aureus in the presence of HLac were estimated. The results can be used together with process data to indicate the range and magnitude of growth and enterotoxin A production during initial stages of cheese production.

Statistical analysis of agronomical factors and weather conditions influencing deoxynivalenol levels in oats in Scandinavia

The relationship between weather data and agronomical factors and deoxynivalenol (DON) levels in oats was examined with the aim of developing a predictive model. Data were collected from a total of 674 fields during periods of up to 10 years in Finland, Norway and Sweden, and included DON levels in the harvested oats crop, agronomical factors and weather data. The results show that there was a large regional variation in DON levels, with higher levels in one region in Norway compared with other regions in Norway, Finland and Sweden. In this region the median DON level was 1000 ng g–1 and the regulatory limit for human consumption (1750 ng g–1) was exceeded in 28% of the samples. In other regions the median DON levels ranged from 75 to 270 ng g–1, and DON levels exceeded 1750 ng g–1 in 3–8% of the samples. Including more variables than region in a multiple regression model only increased the adjusted coefficient of determination from 0.17 to 0.24, indicating that very little of the variation in DON levels could be explained by weather data or agronomical factors. Thus, it was not possible to predict DON levels based on the variables included in this study. Further studies are needed to solve this problem. Apparently the infection and/or growth of DON producing Fusarium species are promoted in certain regions. One possibility may be to study the species distribution of fungal communities and their changes during the oats cultivation period in more detail.