| 1. |
- Blixt, Y., et al.
(författare)
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Interlaboratory random amplified polymorphic DNA typing of Yersinia enterocolitica and Y. enterocolitica-like bacteria
- 2003
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Ingår i: International Journal of Food Microbiology. - 0168-1605 .- 1879-3460. ; 83:1, s. 15-26
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Tidskriftsartikel (refereegranskat)abstract
- A random amplified polymorphic DNA (RAPD) protocol was developed for interlaboratory use to discriminate food-borne Yersinia enterocolitica O:3 from other serogroups of Y. enterocolitica and from Y. enterocolitica-like species. Factors that were studied regarding the RAPD performance were choice of primers and concentration of PCR reagents (template DNA, MgCl 2, primer and Taq DNA polymerase). A factorial design experiment was performed to identify the optimal concentrations of the PCR reagents. The experiment showed that the concentration of the PCR reagents tested significantly affected the number of distinct RAPD products. The RAPD protocol developed was evaluated regarding its discrimination ability using 70 different Yersinia strains. Cluster analysis of the RAPD patterns obtained revealed three main groups representing (i) Y. pseudotuberculosis, (ii) Y. enterocolitica and (iii) Y. kristensenii, Y. frederiksenii, Y. intermedia and Y. ruckeri. Within the Y. enterocolitica group, the European serovar (O:3) and the North American serovar (O:8) could be clearly separated from each other. All Y. enterocolitica O:3 strains were found in one cluster which could be further divided into two subclusters, representing the geographical origin of the isolates. Thus, one of the subclusters contained Y. enterocolitica O:3 strains originating from Sweden, Finland and Norway, while Danish and English O:3 strains were found in another subcluster together with O:9 and O:5,27 strains. The repeatability (intralaboratory) and reproducibility (interlaboratory) of the RAPD protocol were tested using 15 Yersinia strains representing different RAPD patterns. The intralaboratory and the interlaboratory studies gave similarity coefficients of the same magnitude (generally >70%) for the individual strains. In the present study, it was shown that interreproducible RAPD results could be achieved by appropriate optimisation of the RAPD protocol. Furthermore, the study reflects the heterogeneous genetic diversity of the Y. enterocolitica species. © 2002 Elsevier Science B.V. All rights reserved.
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| 2. |
- Dahlenborg, Maria, et al.
(författare)
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Prevalence of Clostridium botulinum types B, E, and F in faecal samples from Swedish cattle.
- 2003
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Ingår i: International Journal of Food Microbiology. - 0168-1605 .- 1879-3460. ; 82:2, s. 105-110
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Tidskriftsartikel (refereegranskat)abstract
- Faeces were collected from 60 cows at three slaughterhouses situated in southern and central Sweden. The faecal samples were collected during two sampling periods over the year, summer and winter. All samples were analysed for the presence of Clostridium botulinum spores, according to a combined selection and enrichment PCR procedure. One PCR assay was specific for part of the type B neurotoxin gene, while the other assay was specific for both type E and F neurotoxin genes. The prevalence of C. botulinum in Swedish cattle was established to be 73% for non-proteolytic type B and less than 5% for types E and F. Twenty-eight (64%) of the positive faecal samples had a spore load of less than 4 spores/g. Statistical analysis (ANOVA) showed that seasonal variation (summer and winter) had a significant effect on the prevalence of C. botulinum type B in cattle, whereas the effect of geographical location of rearing of the cattle (southern and central Sweden) was less significant.
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| 3. |
- Knutsson, Rickard, et al.
(författare)
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Evaluation of selective enrichment PCR procedures for Yersinia enterocolitica.
- 2002
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Ingår i: International Journal of Food Microbiology. - 0168-1605 .- 1879-3460. ; 73:1, s. 35-46
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Tidskriftsartikel (refereegranskat)abstract
- Four enrichment PCR protocols for detecting unlysed cells of pathogenic Yersinia enterocolitica were studied. First, the probability of detecting Y. enterocolitica cells of known concentrations by a multiplex PCR assay was determined, and it was found to follow a logistic regression model. From this model, the probability of detecting Y enterocolitica at a specific concentration could be estimated; for example, the detection probability of 10(4) CFU/ml was estimated to be 85.4%. The protocols were evaluated on enrichment cultures inoculated with 10(2) CFU/ml Y. enterocolitica and 10(2)-10(6) CFU/ml of a defined background flora. For each protocol, the time for sample withdrawal and the presence of background flora were studied with respect to PCR detection. The optimal point in time of sample withdrawal was found to be different for each protocol employed. Early detection was favoured by concentrating the target cells, and the most rapid PCR detection of Y. enterocolitica was achieved with enrichment in Yersinia-PCR-compatible-enrichment (YPCE) medium for 3 h at 25 degrees C, followed by a centrifugation prior to PCR analysis. For detection of Y. enterocolitica in the presence of high concentrations (10(6) CFU/ml) of background flora, a long incubation time followed by density centrifugation and a dilution step was most successful. The protocol that gave the most reliable PCR detection in the presence of 10(6) CFU/ml background flora included 24 h incubation in Yersinia-selective-enrichment (YSE) broth at 25 degrees C, followed by Percoll density centrifugation, and a 100 times dilution prior to PCR analysis.
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| 4. |
- Aronsson, Kristina, et al.
(författare)
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Growth of pulsed electric field exposed Escherichia coli in relation to inactivation and environmental factors
- 2004
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Ingår i: International Journal of Food Microbiology. - 0168-1605 .- 1879-3460. ; 93:1, s. 1-10
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Tidskriftsartikel (refereegranskat)abstract
- Pulsed electric fields (PEF) have been proven to inactivate microorganisms during nonthermal conditions and have the potential to replace thermal processing as a method for food preservation. However, there is a need to understand the recovery and growth of survivors and potentially injured microorganisms following PEF processing. The purpose of this investigation was to study the growth of Escherichia coli at 10°C following exposure to electrical field strengths (15, 22.5 and 30 kV/cm) in relation to inactivation and the amount of potentially sublethally injured cells. One medium was used as both a treatment medium and an incubation medium, to study the influence of environmental factors on the inactivation and the growth of the surviving population. The pH (5.0, 6.0 and 7.0) and water activity (1.00, 0.985 and 0.97) of the medium was varied by adding HCl and glycerol, respectively. Growth was followed continuously by measuring the optical density. The time-to-detection (td) and the maximum specific growth rate (?max) were calculated from these data. Results showed that the PEF process did not cause any obvious sublethal injury to the E. coli cells. The number of survivors was a consequence of the combination of electrical field strength and environmental factors, with pH being the most prominent. Interestingly, the ?max of subsequent growth was influenced by the applied electrical field strength during the process, with an increased ?max at more intense electrical field strengths. In addition, the ?max was also influenced by the pH and water activity. The td, which could theoretically be considered as an increase in shelf life, was found to depend on a complex correlation between electrical field strength, pH and water activity. That could be explained by the fact that the td is a combination of the number of survivors, the recovery of sublethal injured cells and the growth rate of the survivors. © 2003 Published by Elsevier B.V.
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| 5. |
- Beuchat, L. R., et al.
(författare)
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Performance of mycological media in enumerating desiccated food spoilage yeasts : an interlaboratory study
- 2001
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Ingår i: International Journal of Food Microbiology. - : Elsevier. - 0168-1605 .- 1879-3460. ; 70:1-2, s. 89-96
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Tidskriftsartikel (refereegranskat)abstract
- Dichloran 18% glycerol agar (DG18) was originally formulated to enumerate nonfastidious xerophilic moulds in foods containing rapidly growing Eurotium species. Some laboratories are now using DG18 as a general purpose medium for enumerating yeasts and moulds, although its performance in recovering yeasts from dry foods has not been evaluated. An interlaboratory study compared DG18 with dichloran rose bengal chloramphenicol agar (DRBC), plate count agar supplemented with chloramphenicol (PCAC), tryptone glucose yeast extract chloramphenicol agar (TGYC), acidified potato dextrose agar (APDA), and orange serum agar (OSA) for their suitability to enumerate 14 species of lyophilized yeasts. The coefficient of variation for among-laboratories repeatability within yeast was 1.39% and reproducibility of counts among laboratories was 7.1%. The order of performance of media for recovering yeasts was TGYC > PCAC = OSA > APDA > DRBC > DG18. A second study was done to determine the combined effects of storage time and temperature on viability of yeasts and suitability of media for recovery. Higher viability was retained at - 18 degreesC than at 5 degreesC or 25 degreesC for up to 42 weeks, although the difference in mean counts of yeasts stored at - 18 degreesC and 25 degreesC was only 0.78 log(10) cfu/ml of rehydrated suspension. TGYC was equal to PCAC and superior to the other four media in recovering yeasts stored at - 18 degreesC, 5 degreesC, or 25 degreesC for up to 42 weeks. Results from both the interlaboratory study and the storage study support the use of TGYC for enumerating desiccated yeasts. DG18 is not recommended as a general purpose medium for recovering yeasts from a desiccated condition.
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| 6. |
- Lund, Bodil, et al.
(författare)
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Gastrointestinal transit survival of an Enterococcus faecium probiotic strain administered with or without vancomycin
- 2002
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Ingår i: International Journal of Food Microbiology. - 0168-1605 .- 1879-3460. ; 77:1-2, s. 109-115
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Tidskriftsartikel (refereegranskat)abstract
- The primary aim of this study was to evaluate if an ingested probiotic, containing viable Enterococcus faecium could survive gastrointestinal transit and if so, correlate the amount of the recovered probiotic strain with the host's own enterococci. The second aim was to investigate if simultaneous vancomycin intake influenced the survival and persistence of the probiotic strain and the stability of endogenous enterococci strains. Twenty healthy volunteers were given the probiotic product once daily for 10 days. Half of the subjects were simultaneously given vancomycin. Isolates of E. faecium strains were genotypically or phenotypically analysed with pulsed-field gel electrophoresis (PFGE) and the PhenePlate(TM) system, respectively. In eight of the ten volunteers given only the probiotic, the ingested E. faecium could be detected on day 10, while in none on day 31. From subjects given both probiotic and vancomycin no ingested E. faecium could be detected on day 10 or day 31. The estimated amount of ingested E. faecium recovered from faeces on day 10 ranged from 1.2 x 10(3) to 4.2 x 10(6) colony forming units per gram faeces, which in several cases were a substantial part of the total amount of E. faecium. The E. faecium isolated before probiotic plus vancomycin administration showed no close relationship to the ones isolated 3 weeks after ceased intake in any subjects. In conclusion, the ingested E. faecium strain can survive gastrointestinal transit. After intake, the E. faecium probiotic strain might become a large part of the total E, faecium population. The occurrence of the probiotic strain in the human gut seems to be transient after intake stop. Re-colonization of E. faecium after simultaneous probiotic plus vancomycin intake occurs mainly with strains without close genetic relationship to the strains harboured before treatment or to the ingested E. faecium strain.
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| 7. |
- Olsson, J., et al.
(författare)
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Detection and quantification of ochratoxin A and deoxynivalenol in barley grains by GC-MS and electronic nose
- 2002
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Ingår i: International Journal of Food Microbiology. - : Elsevier. - 0168-1605 .- 1879-3460. ; 72:3, s. 203-214
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Tidskriftsartikel (refereegranskat)abstract
- Mycotoxin contamination of cereal grains can be detected and quantified using complex extraction procedures and analytical techniques. Normally, the grain odour, i.e. the presence of non-grain volatile metabolites, is used for quality classification of grain. We have investigated the possibility of using fungal volatile metabolites as indicators of mycotoxins in grain. Ten barley samples with normal odour, and 30 with some kind of off-odour were selected from Swedish granaries. The samples were evaluated with regard to moisture content, fungal contamination, ergosterol content, and levels of ochratoxin A (OA) and deoxynivalenol (DON). Volatile compounds were also analysed using both an electronic nose and gas chromatography combined with mass spectrometry (GC-MS). Samples with normal odour had no detectable ochratoxin A and average DON contents of 16 mug kg(-1) (range 0-80), while samples with off-odour had average OA contents of 76 mug kg(-1) (range 0-934) and DON contents of 69 mug kg(-1) (range 0-857). Data were evaluated by multivariate data analysis using projection methods such as principal component analysis (PCA) and partial least squares (PLS). The results show that it was possible to classify the OA level as below or above the maximum limit of 5 mug kg(-1) cereal grain established by the Swedish National Food Administration, and that the DON level could be estimated using PLS. Samples with OA levels below 5 mug kg(-1) had higher concentration of aldehydes (nonanal, 2-hexenal) and alcohols (1-penten-3-ol, 1-octanol). Samples with OA levels above 5 mug kg(-1) had higher concentrations of ketones (2-hexanone, 3-octanone). The GC-MS system predicted OA concentrations with a higher accuracy than the electronic nose, since the GC-MS misclassified only 3 of 37 samples and the electronic nose 7 of 37 samples. No correlation was found between odour and OA level, as samples with pronounced or strong off-odours had OA levels both below and above 5 mug kg(-1). We were able to predict DON levels in the naturally contaminated barley samples using the volatile compounds detected and quantified by either GC-MS or the electronic nose. Pentane, methylpyrazine, 3-pentanone, 3-octene-2-ol and isooctylacetate showed a positive correlation with DON, while ethylhexanol, pentadecane, toluene, 1-octanol, 1-nonanol, and 1-heptanol showed a negative correlation with DON. The root mean square error of estimation values for prediction of DON based on GC-MS and electronic nose data were 16 and 25 mug kg(-1) respectively.
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| 8. |
- Olsson, J., et al.
(författare)
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Volatiles for mycological quality grading of barley grains : determinations using gas chromatography-mass spectrometry and electronic nose
- 2000
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Ingår i: International Journal of Food Microbiology. - : Elsevier. - 0168-1605 .- 1879-3460. ; 59:3, s. 167-178
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Tidskriftsartikel (refereegranskat)abstract
- The possibility of using an electronic nose or gas chromatography combined with mass spectrometry (GC-MS) to quantify ergosterol and colony forming units (CFU) of naturally contaminated barley samples was investigated. Each sample was split into three parts for (i) ergosterol and CFU analysis, (ii) measurements with the electronic nose and (iii) identification of volatiles collected on an adsorbent with a GC-MS system. Forty samples were selected after sensory analysis to obtain 10 samples with normal odour and 30 with some degree of off-odour. The data set of volatile compounds and the data collected from the electronic nose were evaluated by multivariate analyse techniques. SIMCA classification (soft independent modelling of class analogy) was used for objective evaluation of the usefulness of the data from the GC-MS or electronic nose measurements for classification of grain samples as normal or with off-odour. The main volatile compounds of grain with normal odour were 2-hexenal, benzaldehyde and nonanal, while 3-octanone, methylheptanone and trimethylbenzene were the main volatile compounds of grain with off-odours. Using data from the electronic nose three samples of 40 were misclassified, while data analysis of the volatile compounds detected with the GC-MS, led to six misclassified samples. Regression models (partial least-squares, PLS) were built to predict ergosterol- and CFU-levels with data from the GC-MS or electronic nose measurements. PLS models based on both GC-MS and electronic nose data could be used to predict the ergosterol levels with high accuracy and with low root mean square error of prediction (RMSEP). CFU values from naturally infected grain could not be predicted with the same degree of confidence.
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| 9. |
- Suihko, M.-L., et al.
(författare)
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Characterization of Listeria monocytogenes isolates from the meat, poultry and seafood industries by automated ribotyping
- 2002
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Ingår i: International Journal of Food Microbiology. - 0168-1605 .- 1879-3460. ; 72:42006, s. 137-146
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Tidskriftsartikel (refereegranskat)abstract
- A total of 564 Listeria monocytogenes isolates were characterized by automated ribotyping. The samples were taken from equipment, personnel and the environment after cleaning procedures and during food processing, as well as from raw materials and products from six meat, two poultry and five seafood processing plants located in the Faroe Islands, Finland, Iceland, Norway and Sweden. Altogether, 25 different ribotypes (RTs) were generated. Two RTs occurred in the samples from all three food sectors-meat, poultry and seafood. Four RTs occurred in meat and poultry plant samples and other four RTs occurred in meat and seafood plant samples. Five RTs occurred only in meat plant samples, five only in poultry plant samples and five only in seafood plant samples. Eight of the thirteen plants had their own in-house L. monocytogenes ribotype. There was geographical differences between the RTs, but no correlation between RTs and food sectors was detected. The discrimination power of automated ribotyping was satisfactory to trace the contamination sources in the food processing plants clearly indicating the sites at which improved cleaning procedures were necessary. In addition, it was possible to screen a large number of isolates with two instruments located at different institutes and to make a reliable combination of the results. Copyright © 2002 Elsevier Science B.V.
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| 10. |
- Söderström, Charlotte, et al.
(författare)
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Use of an electronic tongue to analyze mold growth in liquid media
- 2003
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Ingår i: International Journal of Food Microbiology. - 0168-1605 .- 1879-3460. ; 83:3, s. 253-261
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Tidskriftsartikel (refereegranskat)abstract
- The feasibility of employing an electronic tongue to measure the growth of mold in a liquid medium was studied. We used the electronic tongue developed at Linköping University, which is based on pulsed voltammetry and consists of an array of different metal electrodes. Instead of focusing on a single parameter, this device provides information about the condition or quality of a sample or process. Accordingly, the data obtained are complex, and multivariate methods such as principal component analysis (PCA) or projection to latent structures (PLS) are required to extract relevant information. A gas chromatographic technique was developed to measure ergosterol content in mold biomass and was subsequently used as a reference method to investigate the ability of the electronic tongue to measure the growth of mold in liquid media. The result shows that the electronic tongue can monitor mold growth in liquids. In PLS analysis, the electronic tongue signals correlate well with the amount of ergosterol in the mold biomass as well as the microbially induced changes in the pH of the medium. © 2002 Elsevier Science B.V. All rights reserved.
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