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  • Kylä-Puhju, Maria; Ruusunen, Marita; Puolanne, Eero (Elsevier, 2005)
    The activity of glycogen debranching enzyme (GDE) was studied in relation to pH value and temperature in porcine masseter and longissimus dorsi muscles. A glycogen limit dextrin was used as the substrate for GDE, and the enzyme was derived from raw meat extracts. In both muscles, the pH only weakly affected on activity of GDE at the pH values found in carcasses post-slaughter. However, the activity of GDE decreased strongly (P < 0.001) when the temperature decreased from values of 39 °C and 42 °C found just after slaughter to values of 4 °C and 15 °C found during cooling. In both muscles the activity of GDE began to fall at temperatures below 39 °C and was almost zero when the temperature decreased to below 15 °C. Thus, the activity of GDE may control the rate of glycogenolysis and glycolysis, but does not block rapid glycolysis and pH decrease when the temperature is high. This may be important in PSE meat, where the pH decreases rapidly at high temperatures, but rapid cooling could limit the activity of GDE and thus glycogenolysis. As expected, GDE was more active in the light longissimus dorsi muscle than in the dark masseter muscle.
  • Rahkio, T.; Korkeala, H. (International Association for Food Protection, 1997)
  • Lyytikäinen, O.; Autio, T.; Maijala, R.; Ruutu, P.; Honkanen-Buzalski, T.; Miettinen, M.; Hatakka, M.; Mikkola, J.; Anttila, V.-J.; Johansson, T.; Rantala, L.; Aalto, T.; Korkeala, H.; Siitonen, A. (University of Chicago Press, 2000)
    In February 1999, an outbreak of listeriosis caused by Listeria monocytogenes serotype 3a occurred in Finland. All isolates were identical. The outbreak strain was first isolated in 1997 in dairy butter. This dairy began delivery to a tertiary care hospital (TCH) in June 1998. From June 1998 to April 1999, 25 case patients were identified (20 with sepsis, 4 with meningitis, and 1 with abscess; 6 patients died). Patients with the outbreak strain were more likely to have been admitted to the TCH than were patients with other strains of L. monocytogenes (60% vs. 8%; odds ratio, 17.3; 95% confidence interval, 2.8136.8). Case patients admitted to the TCH had been hospitalized longer before cultures tested positive than had matched controls (median, 31 vs. 10 days; P = .008). An investigation found the outbreak strain in packaged butter served at the TCH and at the source dairy. Recall of the product ended the outbreak.
  • Nevas, M.; Korhonen, A.-R.; Lindström, M.; Turkki, P.; Korkeala, H. (International Association for Food Protection, 2004)
  • Haapoja, A.; Korkeala, H. (BioMed Central, 1984)
  • Keto-Timonen, R; Tolvanen, R; Lunden, J; Korkeala, H (2007)
    Contamination routes of Listeria monocytogenes were examined in a chilled food processing plant that produced ready-to-eat and ready-to-reheat meals during an 8-year period by amplified fragment length polymorphism (AFLP) analysis. A total of 319 L. monocytogenes isolates were recovered from raw materials (n=18), the environment (n=77), equipment (n=193), and products (n=31), and 18 different AFLP types were identified, five of which were repeatedly found to be persistent types. The three compartments (I to III) of the plant showed markedly different contamination statuses. Compartment I, which produced cooked meals, was heavily contaminated with three persistent AFLP types. AFLP type A1 dominated, and it comprised 93% of the isolates of the compartment. Compartment II, which produced uncooked chilled food, was contaminated with four persistent and five nonpersistent AFLP types. The equipment of compartment III, which produced cooked ready-to-reheat meals, was free of contamination. In compartments that produced cooked meals, the cleaning routines, product types, and lack of compartmentalization seemed to predispose production lines to persistent contamination. The most contaminated lines harbored L. monocytogenes in coolers, conveyors, and packing machines. Good compartmentalization limited the flow of L. monocytogenes into the postheat -treatment area and prevented the undesired movement of equipment and personnel, thus protecting the production lines from contamination. In compartment II, grated cheese was shown to cause product contamination. Therefore, special attention should be paid to continuous quality control of raw ingredients when uncooked ready-to-eat foods are produced. In compartment II, reconstruction of the production line resulted in reduced prevalence rates of L. monocytogenes and elimination of two persistent AFLP types.
  • Pouttu, Petri; Puolanne, Eero (Elsevier, 2004)
    An attempt was made to determine the water-binding capacity of each individual trimming in a multicomponent system. Three types of experimental cooked sausages (finely chopped luncheon sausage, coarsely chopped sausage and ring sausage with potato starch) were made of five different meat trimmings: two pork trimmings and two beef trimmings, and one beef trimming used as a replacer. The water-binding was determined by the Tuominen-Honkavaara method by stepwise addition of water (basic formulation and four water additions) to the formulations and determining the firmness by a consistometer. The water-binding of each trimming was obtained by replacing the trimming by an additional trimming. A total of 3 sausage types x 5 meat trimmings x 5 water levels giving 75 experimental batches of five kg each were made. The average water-binding values of the same meat trimming combination in each sausage type were practically the same, and therefore the total averages for the same meat trimming combinations of each of the three sausage types were used for the subsequent calculations. The determination of the water-binding values of the meat trimmings were solved by forming five equations with four unknowns each, and then solving the unknowns using Microsoft Excel's ‘Solver’ function. By this procedure it was possible to determine the water-binding of individual meat trimmings in sausage systems. This procedure can be used for the determination of the technological properties of meats for linear programming.
  • Augustin, M.; Ali-Vehmas, T.; Atroshi, F. (Canadian Society for Pharmaceutical Sciences, 2004)
    PURPOSE: Microbial biofilm has become difficult to control by antibiotic and biocide regimes that are effective against suspended bacteria. Their colonization of surfaces can be a problem and is generally controlled through cleaning and disinfection. This study was undertaken to examine the efficacy of the disinfectants including Bio-Ow, Econase CE, Gamanase GC 140, IndiAge 44L, Mannanase AMB, Multifect P-3000, Neutrase, Pandion, Paradigm, Pectinex Ultra SP-L, Promozyme, Resinase A2X, Spezyme AA300, Spezyme GA300 and Vinozym EC, and the proteinase against bacterial biofilms. METHODS: The effectiveness of 20 commercial disinfectants against Pseudomonas aeruginosa (P. aeruginosa) biofilms using a fluorometric technique was examined. Additionally the disinfectants were also tested against Lactobacillus bulgaricus (L. bulgaricus), Lactobacillus lactis (L. lactis) and Streptococcus thermophilus (S. thermophilus) isolates using microtitration tray based turbidimetric techniques. Escherichia coli (E. coli) was used as the test bacteria in the fluorometric control method. RESULTS: Among the first group of the enzymatic cleaning agents tested, four disinfectants (Pandion, Resinase A2X, Spezyme GA300 and Paradigm) were the most potent against bacterial biofilms after 30 min incubation time (residual bacterial count less than 10(3) CFU (colony forming units)/ml). However, only Resinase A2X and Paradigm showed a good effect on bacterial biofilms after 15 min incubation time. Proteinase disinfectants (alkalase, chymotrypsin, cryotin and krilltrypsin) from the second group of the disinfectants showed a good effect against P. aeruginosa biofilm when tested in the absence of milk. The performance of the disinfectants was reduced in the presence of milk. The minimum inhibitory concentration (MIC) of the cleaning agents was determined as the lowest concentration inhibiting bacterial growth. The MIC was tested on Lactobacillus bulgaricus (L. bulgaricus), Lactobacillus lactis (L. lactis) and Streptococcus thermophilus (S. thermophilus) isolates. The minimum inhibitory concentrations (MIC) for Paradigm against S. thermophilus and L. Lactis were lower than L. Bulgaricus. Whereas, the MIC of Pandion against L. bulgaricus was lower than MIC against L. lactis. Resinase A2X had no inhibitory effect on bacterial growth when the concentration was less than or equal to 2.4 mg/ml and Spezyme GA 300 concentration less than or equal to 7.3 mg/ml. Minimum inhibitory concentration of Pandion against L. bulgaricus was 2.7 microg/ml and against L. lactis 5.3 microg/ml. Growth of S. thermophilus was inhibited in all concentration of Pandion tested. CONCLUSIONS: The choice of disinfectant or cleaning agent along with the optimum concentration and the time of action is very important when destroying microbes. It is also important that the resistances of microbes to different disinfectants and cleaning agents be taken into account when planning the cleaning process
  • Nuorti, J.P.; Niskanen, T.; Hallanvuo, S.; Mikkola, J.; Kela, E.; Hatakka, M.; Fredriksson-Ahomaa, M.; Lyytikäinen, O.; Siitonen, A.; Korkeala, H.; Ruutu, P. (University of Chicago Press, 2004)
    Background. The vehicles and sources of Yersinia pseudotuberculosis infection are unknown. In Finland, clinical microbiology laboratories routinely report Y. pseudotuberculosis isolations and submit isolates for serotype analysis. In October 1998, the number of serotype O:3 infections increased markedly. Methods. Case patients with culture-confirmed Y. pseudotuberculosis O:3 infection were identified by use of laboratory-based surveillance. We conducted a population-based case-control study. Healthy community control subjects were matched by age, sex, and postal code. Isolates were subtyped by pulsed-field gel electrophoresis (PFGE). Results. Nationwide, 47 case patients were identified (age range, 277 years; median, 19 years). One patient with bacteremia died; 5 underwent appendectomies. We enrolled 38 case patients and 76 control subjects in the case-control study. Seventy-one percent of case patients and 42% of control subjects reported having eaten iceberg lettuce (matched odds ratio, 3.8; 95% confidence interval, 1.39.4); a dose-response relationship was found for increasing frequency of consumption. Of the 27 isolates obtained from case patients and tested in the analysis, all had indistinguishable PFGE patterns. Four lunch cafeterias that had served iceberg lettuce were associated with clusters of case patients. The lettuce was traced back to originating farms. Conclusions. Iceberg lettuce was implicated as the vehicle of a widespread foodborne Y. pseudotuberculosis outbreak. Ongoing laboratory-based surveillance and serotype analysis were essential in the rapid detection of infection. Cases of yersiniosis, which appear to be sporadic, may be part of unrecognized outbreaks caused by contaminated fresh produce.
  • Hyytiä, E.; Hielm, S.; Björkroth, J.; Korkeala, H. (American Society for Microbiology, 1999)
    The genetic biodiversity of Clostridium botulinum type E strains was studied by pulsed-field gel electrophoresis (PFGE) with two macrorestriction enzymes (SmaI-XmaI and XhoI) and by randomly amplified polymorphic DNA (RAPD) analysis with two primers (OPJ 6 and OPJ 13) to characterize 67 Finnish isolates from fresh fish and fishery products, 15 German isolates from farmed fish, and 10 isolates of North American or North Atlantic origin derived mainly from different types of seafood. The effects of fish species, processing, and geographical origin on the epidemiology of the isolates were evaluated. Cluster analysis based on macrorestriction profiles was performed to study the genetic relationships of the isolates. PFGE and RAPD analyses were combined and resulted in the identification of 62 different subtypes among the 92 type E isolates analyzed. High genetic biodiversity among the isolates was observed regardless of their source. Finnish and North American or North Atlantic isolates did not form distinctly discernible clusters, in contrast with the genetically homogeneous group of German isolates. On the other hand, indistinguishable or closely related genetic profiles among epidemiologically unrelated samples were detected. It was concluded that the high genetic variation was probably a result of a lack of strong selection factors that would influence the evolution of type E. The wide genetic biodiversity observed among type E isolates indicates the value of DNA-based typing methods as a tool in contamination studies in the food industry and in investigations of botulism outbreaks.
  • Pösö, A. Reeta; Puolanne, Eero (Elsevier, 2005)
    Oxidative energy production is by far dominant in living animal muscles, with the exception the short periods of severe stress, where the aerobic capacity is exceeded, and formation of large amounts of lactic acid will take place. Energy consumption in muscle cells continues post mortem with formation of large amounts of lactate and formation of protons, because the aerobic processes for energy production are not available. Post mortem, the fall in pH is delayed only by buffering capacity of the muscle fibres. In living animals, in addition to buffering capacity, both respiration and transport of lactate and protons out of the muscle fibres by monocarboxylate transporters participate in the regulation of muscle fibre pH which never falls as low as the ultimate pH of the meat. Understanding the regulation of pH in muscle is important both for the welfare of living animals and from the technological point of view as a factor influencing meat quality.
  • Sarkkola, Sakari; Alenius, Virpi; Hökkä, Hannu; Laiho, Raija; Päivänen, Juhani; Penttilä, Timo (NRC Canada, 2003)
    Size-structural dynamics of naturally established Norway spruce (Picea abies (L.) Karst.) stands growing on peatlands drained for forestry were investigated. The study was based on modelling of diameter at breast height (DBH) distributions of repeatedly measured stands in southern Finland. The Weibull function was used to parameterize the DBH distributions and mixed linear models were constructed to characterize the impacts of different ecological factors on stand dynamics. Initially, the positive skewness of the DBH distributions increased after drainage as a result of increases in stem numbers and a reduction in mean diameters. Simultaneously, the size inequality among trees increased. These changes were due to regeneration and (or) ingrowth and indicated only little competition from the larger trees. Subsequently, the DBH distributions changed from positively skewed to normal and finally to negatively skewed resulting from tree growth and a reduction in the number of small DBH trees. This indicated increased asymmetric intertree competition. Size inequality did not change during this later stage in stand development, suggesting a concurrent component of symmetric competition. Thinnings had little impact on DBH distribution trends. The observed stand dynamics allow the allocation of growth resources to the desired crop component by appropriate silvicultural treatments.
  • Lyhs, Ulrike; Björkroth, Johanna; Korkeala, Hannu (Elsevier, 1999)
    A total of 405 lactic acid bacteria (LAB) isolated from spoiled, vacuum-packaged, salted, sodium nitrite- or potassium nitrate-treated, cold-smoked rainbow trout stored at 48C or 88C were characterised and identified using a molecular method. The isolates were initially classified according to their restriction endonuclease profiles using HindIII and EcoRI restriction endonucleases and further characterised by rRNA gene restriction patterns (ribotypes). Numerical analysis of these ribopatterns was performed together with 19 reference LAB strain patterns in order to identify the isolates to species level. The strains were divided with HindIII and EcoRI ribopatterns into ten and nine clusters at the similarity level of 65% and 50%, respectively. The Leuconostoc-clusters and the Lb. sakei /Lb. curvatus-clusters formed the two main groups. Only one isolate was identified as Lactobacillus plantarum and no Carnobacterium strains were discovered. For both enzymes, the 35 isolates possessing six individual ribotypes and forming five clusters could not be identified further with the reference strains used. The relative proportion of Leuconostoc mesenteroides subsp. mesenteroides was higher in all samples stored at 48C. Most of the Leuconostoc citreum were found in the samples stored at 88C, and particularly in the nitrite-treated samples.
  • Hyytiä-Trees, Eija; Lyhs, Ulrike; Korkeala, Hannu; Björkroth, Johanna (Elsevier, 1999)
    Eighteen earlier well characterised Lactobacillus sake strains representing different slime production capabilities in vacuum-packaged meat products were analysed using repetitive element sequence-based PCR (rep-PCR). The single primers BOXA1R and RW3A and the primer pair REP1R-Dt & REP2R-Dt were evaluated for their applicability in L. sake genotyping. The five different patterns produced by RW3A were least revealing, with the discriminatory power equalling to ribotyping. BOXA1R and REP-primer pair both produced six different banding patterns and the combination of these results yielded seven different rep-types. Rep-PCR was concluded to have approximately the same discriminatory power as randomly amplified polymorphic DNA (RAPD) analysis, but it was inferior to pulsed-field gel electrophoresis (PFGE). How-ever, if the results of rep-PCR and RAPD were combined, the discrimination was comparable PFGE, with the exception that within Ribogroup I the non-slime-producing strains were not distinguished from weak slime producers. It was concluded that the combination of the two PCR-based typing techniques, rep-PCR and RAPD, would be a valuable tool in large scale contamination studies at the meat processing plants, since results can be obtained rapidly and fewer isolates need further analysis by PFGE.
  • Santos, Eva M.; Jaime, Isabel; Rovira, Jordi; Lyhs, Ulrike; Korkeala, Hannu; Björkroth, Johanna (Elsevier, 2005)
    A total of 176 lactic acid bacteria (LAB) isolated from a typical Spanish blood sausage called “morcilla de Burgos” were identified by means of phenotypic characteristics and 16S rDNA RFLP (ribotyping). LAB were isolated from “morcilla” of different producers and in different storage periods, that includes unpackaged, vacuum and modified atmosphere packaged “morcilla” and vacuum packed and pasteurised “morcilla”. The knowledge of specific spoilage bacteria of ”morcilla de Burgos” will be useful to design new preservation methods to extent the shelf-life of this product. Identification made according to phenotypic and biochemical characteristics shows the majority of the isolates were heterofermentative LAB (93.2%) and eight different bacterial groups could be distinguished (A-G). W. viridescens was the main species detected (42%). In addition, Leuconostoc spp. (23.9%), W. confusa (11.4%) and Lactobacillus fructosus (5.7%) species were found. Few strains were phenotypically missidentified as Lb. sanfrancisco, Pediococcus spp., Lb. sakei/curvatus and Carnobacterium spp. and 11 strains remained unknown. Most of the leuconostocs were identified as Lc. mesenteroides and Lc. carnosum species. Ribotyping shows a quite good correlation with phenotypic methods, although it has been possible to identify 15 different clusters. W. viridescens and leuconostocs were also the predominant LAB. Strains identified as W. confusa by phenotypic characteristics were resolved in W. confusa and W. cibaria by ribotyping. Neither Carnobacterium piscicola nor Lb. sanfrancisco were identified by means of genotypic method. All Lb. fructosus strains and some more included in different phenotypic groups (17 strains in total) could not be associated with any reference strain (cluster VII). Although some discrepancies exists the combination of phenotypic and genotypic methods led to a better identification and characterization of the strains isolated from “morcilla de Burgos”.
  • Ridell, J.; Siitonen, A.; Paulin, L.; Lindroos, O.; Korkeala, H.; Albert, M.J. (American Society for Microbiology, 1995)
    Hafnia alvei strains which possess the attachment-effacement gene (eaeA) may have clinical importance as new diarrhea-causing pathogens and should therefore be differentiated from other H. alvei strains. We characterized diarrheal H. alvei strains, which were positive in the PCR test for the eaeA gene, using biochemical tests not routinely used for identification of members of the family Enterobacteriaceae, and compared them with eaeA-negative strains isolated from different clinical and nonclinical sources to find characteristics useful for identification. Random amplified polymorphic DNA (RAPD)-PCR and partial sequencing of the 16S rRNA gene were utilized to study the genetic diversity of the isolates. The eaeA-positive strains were found to have many characteristic biochemical properties. Negative reactions in the 2- ketogluconate and histidine assimilation tests and a positive reaction in the 3-hydroxybenzoate assimilation test may be useful in routine diagnostics. Nearly identical RAPD-PCR profiles and identical 353-bp fragments of the 16S rRNA genes indicated little genetic diversity among the eaeA-positive strains. The low level of homology (92%) in the partial 16S rRNA genes of eaeA-positive and -negative H. alvei strains raises questions about the taxonomic positioning of eaeA-positive H. alvei.
  • Björkroth, Johanna; Geisen, Rolf; Schillinger, Ulrich; Weiss, Norbert; De Vos, Paul; Holzapfel, Wilhelm H.; Korkeala, Hannu; Vandamme, Peter (American Society for Microbiology (ASM), 2000)
    Lactic acid bacteria (LAB) associated with gaseous spoilage of modified-atmosphere-packaged, raw, tomatomarinated broiler meat strips were identified on the basis of a restriction fragment length polymorphism (RFLP) (ribotyping) database containing DNAs coding for 16S and 23S rRNAs (rDNAs). A mixed LAB population dominated by a Leuconostoc species resembling Leuconostoc gelidum caused the spoilage of the product. Lactobacillus sakei, Lactobacillus curvatus, and a gram-positive rod phenotypically similar to heterofermentative Lactobacillus species were the other main organisms detected. An increase in pH together with the extreme bulging of packages suggested a rare LAB spoilage type called “protein swell.” This spoilage is characterized by excessive production of gas due to amino acid decarboxylation, and the rise in pH is attributed to the subsequent deamination of amino acids. Protein swell has not previously been associated with any kind of meat product. A polyphasic approach, including classical phenotyping, whole-cell protein electrophoresis, 16 and 23S rDNA RFLP, 16S rDNA sequence analysis, and DNA-DNA reassociation analysis, was used for the identification of the dominant Leuconostoc species. In addition to the RFLP analysis, phenotyping, whole-cell protein analysis, and 16S rDNA sequence homology indicated that L. gelidum was most similar to the spoilage-associated species. The two spoilage strains studied possessed 98.8 and 99.0% 16S rDNA sequence homology with the L. gelidum type strain. DNA-DNA reassociation, however, clearly distinguished the two species. The same strains showed only 22 and 34% hybridization with the L. gelidum type strain. These results warrant a separate species status, and we propose the name Leuconostoc gasicomitatum sp. nov. for this spoilage-associated Leuconostoc species.
  • Lahti P; Heikinheimo A; Johansson T; Korkeala H (2007)
    The prevalences of various genotypes of enterotoxin gene-carrying (cpe-positive) Clostridium perfringens type A in 24 different food poisoning outbreaks were 75% (chromosomal IS1470-cpe), 21% (plasmid-borne IS1470-like-cpe), and 4% (plasmid-borne IS1151-cpe). The results show that C. perfringens type A carrying the plasmid-borne cpe is a common cause of food poisoning.
  • Korkeala, H.; Sorvettula, O.; Mäki-Petäys, O.; Hirn, J. (BioMed Central, 1982)
  • Mäki-Petäys, O.; Korkeala, H.; Alanko, T.; Sorvettula, O. (BioMed Central, 1991)