Template-Type: ReDIF-Article 1.0 Author-Name: Kateřina Kučerová Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Ivana Korbová Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Šárka HORÁČKOVÁ Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Eva Šviráková Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Milada Plocková Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Title: Influence of Enterococci and Lactobacilli on Listeria Abstract: A collection of lactic acid bacteria (38 Enterococcus and 41 Lactobacillus strains) was tested for the antilisterial activity against 15 Listeria spp. strains (two L. monocytogenes, one L. ivanovii and 12 L. innocua strains) using agar spot method. Out of all 79 bacteria only six Enterococcus strains (1/3A, 3/3A, 6/4D, 6/1A, 1282 and EN3) exhibited antilisterial activity against almost all used indicator strains, when their live cells were tested. When their cell free neutralised supernatants (CFNS) were tested against four selected indicator strains (L. innocua Ln-03, Ln-06, Ln-10 and L. monocytogenes CCM5576) only two Enterococcus spp. strains were active - E. faecalis 6/1A strain from raw cow milk of minor interest due to the activity of its CFNS only against L. innocua Ln-06 and thermolability of the compound and E. mundtii 1282 strain from goat raw milk with CFNS active against 13 Listeria spp. strains including L. monocytogenes. E. mundtii 1282 strain produced probably a bacteriocin, because it completely lost the activity after treatment CFNS with proteinase K. Keywords: Enterococcus, Lactobacillus, Listeria, antilisterial activity Journal: Czech Journal of Food Sciences Pages: SII12-SII17 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/676-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/676-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0001.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:676-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Anna Hudecová Author-Workplace-Name: Department of Biochemistry, Nutrition and Food Assessment, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Bratislava, Slovak Republic Author-Name: Ľubomír Valík Author-Workplace-Name: Department of Biochemistry, Nutrition and Food Assessment, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Bratislava, Slovak Republic Author-Name: Denisa Liptáková Author-Workplace-Name: Department of Biochemistry, Nutrition and Food Assessment, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Bratislava, Slovak Republic Title: Quantification of Geotrichum candidum growth in co-culture with lactic acid bacteria Abstract: The growth dynamics of filamentous fungus G. candidum was studied during the co-cultivation with the commercial lactic acid bacteria (LAB) culture Fresco. The experiments were carried out in milk and on the surface of a milk agar at the temperature ranging from 5 to 37°C. Ratkowsky model was used to describe the relationships of the fungal growth rate to the temperature during both, single and co-cultivation with LAB in milk. Simultaneous growth of LAB affected significantly the growth rate of the filamentous fungus. The growth of G. candidum was in average 39% slower in the co-culture than in the single cultivation. LAB pre-inoculated and growing in the solid medium did not show any significant inhibitory effect on the surface growth of G. candidum at all tested temperature. The precise data describing the growth of this cheese yeast-like fungus, G. candidum, may fill a gap in the field of quantitative food mycology and may be used for predicting its behavior in real conditions. Keywords: Geotrichum candidum, lactic acid bacteria, growth modelling Journal: Czech Journal of Food Sciences Pages: SII18-SII27 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/205/2009-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/205/2009-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0002.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:205-2009-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Alžbeta Medveďová Author-Workplace-Name: Department of Nutrition and Food Assessment, Institute of Biochemistry, Nutrition and Health Protection, Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava, Slovak Republic Author-Name: Ľubomír Valík Author-Workplace-Name: Department of Nutrition and Food Assessment, Institute of Biochemistry, Nutrition and Health Protection, Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava, Slovak Republic Author-Name: Adriana Studeničová Author-Workplace-Name: Department of Nutrition and Food Assessment, Institute of Biochemistry, Nutrition and Health Protection, Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava, Slovak Republic Title: The effect of temperature and water activity on the growth of Staphylococcus aureus Abstract: The growth responses of Staphylococcus aureus 2064 as affected by water activity and incubation temperature were studied in two different laboratory media. Growth parameters at temperatures from 7 to 51°C and aw in the range from 1.0 to 0.86 were fitted using Ratkowsky models. The effect of temperature within its whole range on the specific growth rate was modelled by the extended model under the following equation: √µ = 0.0456 (T - Tmin) [1 - e0.447(T - Tmax)]. The water activity values of tested media were adjusted by sodium chloride in the range from aw = 1.0 to 0.86 and experiments were conducted at 15 and 18°C. The growth responses of S. aureus on water activity at 15°C and 18°C in PCA broth and BHI broth was described by simplified Ratkowsky model in the form: √µ = b × aw. Validation of the found relationships confirmed sound fitting of the data and thus the referred results of the isolate originated from ewes' cheese can be used in the growth prediction of S. aureus, reliably. Keywords: Staphylococcus aureus, temperature, water activity, predictive microbiology Journal: Czech Journal of Food Sciences Pages: SII28-SII35 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/204/2009-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/204/2009-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0003.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:204-2009-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Tereza Gelbíčová Author-Workplace-Name: National Institute of Public Health Prague, Brno, Czech Republic Author-Name: Renata Karpíšková Author-Workplace-Name: National Institute of Public Health Prague, Brno, Czech Republic Author-Workplace-Name: University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic Title: Occurrence and characteristics of Listeria monocytogenes in ready-to-eat food from retail market in the Czech Republic Abstract: The study objectives were to test ready-to-eat food from the retail market in the Czech Republic for the presence of L. monocytogenes and, based on typing methods, to investigate probable causes of contamination. A total 2180 samples of ready-to-eat food (meat, dairy, fish, delicatessen and confectionery products and fresh fruit and vegetables) were analysed qualitatively and quantitatively. L. monocytogenes isolates were characterised by serotyping and macrorestriction analysis after digestion with the restriction enzyme AscI. In 2004-2008 L. monocytogenes was most often detected in delicatessen (5.2%), meat (3.4%) and dairy products (1.8%). In the analysed samples, L. monocytogenes was mostly present at counts lower than 102 CFU/g. Only in 2004, higher counts of L. monocytogenes were found in two heat-processed meat products (103 CFU/g). The obtained macrorestriction patterns helped in tracing the source of contamination and routes of the spread of L. monocytogenes in the manufacturing plant and retail market. Keywords: L. monocytogenes, foodstuff, typing methods, hygiene Journal: Czech Journal of Food Sciences Pages: SII3-SII7 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/210/2009-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/210/2009-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0004.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:210-2009-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Zora Šťástková Author-Workplace-Name: University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Author-Name: Sylva Karpíšková Author-Workplace-Name: Czech Collection of Microorganisms, Institute of Experimental Biology, Masaryk University, Brno, Czech Republic Author-Name: Renáta Karpíšková Author-Workplace-Name: University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Author-Workplace-Name: National Institute of Public Health Prague, Brno, Czech Republic Title: Findings of methicillin-resistant strains of Staphylococcus aureus in livestock Abstract: The aim of our study was to determine the occurrence of methicillin resistant Staphylococcus aureus (MRSA) at dairy farms in the Czech Republic. Altogether 1061 samples from 95 farms were examined. The samples analysed were milk (individual and bulk tank milk samples), animal swabs and swabs from the farm environment. In total, 299 S. aureus isolates were obtained, of which 23 were MRSA. These MRSA isolates originated from three farms (13 isolates from farm A and 5 isolates from each of farms B and C). All MRSA isolates carried the mecA gene while none of them carried the genes for PVL, TSST-1 and exfoliatins. Only the isolates from goat farm C were positive for the genes encoding enterotoxins. By SCCmec typing, the strains were classified as community-associated MRSA carrying SCCmec IV or V. This study revealed that animals can be an important source of methicillin resistant staphylococci and represent a potential hazard of further spread. Keywords: MRSA, mecA, Panton-Valentine leukocidin, toxic-shock syndrome toxin-1, staphylococcal enterotoxins, exfoliative toxins, SCCmec, resistance Journal: Czech Journal of Food Sciences Pages: SII36-SII41 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/209/2009-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/209/2009-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0005.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:209-2009-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Jan Hodek Author-Workplace-Name: Department of Molecular Biology, Crop Research Institute, Prague-Ruzyně, Czech Republic Author-Name: Jaroslava Ovesná Author-Workplace-Name: Department of Molecular Biology, Crop Research Institute, Prague-Ruzyně, Czech Republic Author-Name: Ladislav Kučera Author-Workplace-Name: Department of Molecular Biology, Crop Research Institute, Prague-Ruzyně, Czech Republic Title: Interferences of PCR effectivity: importance for quantitative analyse Abstract: Importance of the Polymerase chain reaction (PCR) have already crossed the border of mere target DNA sequence present or absence analysis. For number analyses e.g. Genetically Modified Organisms (GMOs) or gene expression assesment the DNA quantification is demanded. Real-time (or quantitative) PCR is the most used tool for nucleic acids quantification. PCR efficiency has relevant importance on DNA quantification - it should be almost same for each PCR and its value should varied between 90-100%. There are a lot of PCR enhancers and inhibitors well known. We described impact of used DNA solvent and used laboratory plastic on real-time PCR efficiency. Keywords: real-time PCR, DNA quantification, PCR efficiency, GMO analysis Journal: Czech Journal of Food Sciences Pages: SII42-SII49 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/677-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/677-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0006.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:677-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Kateřina Kučerová Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Hana Svobodová Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Štěpán Tůma Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Iva Ondráčková Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Milada Plocková Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Title: Production of biogenic amines by Enterococci Abstract: Enterococci were presented in all tested samples of raw cow milk (six samples) at the level 103-105 CFU/ml, fresh cheeses (five samples) at the level 102-106 CFU/g and semi-hard cheeses (five samples) at the level 103-105 CFU/g. All 33 isolated Enterococcus strains were screened for decarboxylase activity by usage differential growth medium and 20 of them possessed tyrosine decarboxylase activity. A collection of eight strains with the strongest decarboxylase activity were identified by species specific PCR as E. faecium (Z3, Z4, Br4 and 6/4D strains) and E. faecalis (Ž4, 3/3C and 4/1A strains). Enterococcus spp. Z1 strain was not identified at the species level by used methods, but the genus was confirmed by PCR method. Their tyrosin decarboxylase activity was confirmed by TLC and detection of tdc gene. Z1, Z3 and Z4 strains showed also histidine decarboxylase activity on the differential growth medium with histidine, but this activity was evaluated by TLC as a false positive reaction of medium. Keywords: Enterococcus, decarboxylase, tyrosine, histidine, TLC, PCR Journal: Czech Journal of Food Sciences Pages: SII50-SII55 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/673-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/673-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0007.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:673-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Lukáš Valihrach Author-Workplace-Name: Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Kateřina Demnerová Author-Workplace-Name: Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Renata Karpíšková Author-Workplace-Name: National Institute of Public Health Prague, Brno, Czech Republic Author-Name: Ivana Melenová Author-Workplace-Name: Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Title: The expression of selected genes encoding enterotoxins in Staphylococcus aureus strains Abstract: Staphylococcus aureus is an important food-borne pathogen, which produces many toxic substances that cause a variety of illnesses. Some strains of S. aureus produce thermostable enterotoxins that can be responsible for alimentary intoxication. The aim of this work was to establish a protocol for the study of 9 enterotoxin genes expression (sea-sej). First, a method for the detection of genes encoding enterotoxins was established and then a method for the determination of the expression of these genes was optimised, using a range of the PCR techniques (multiplex, touchdown and real-time). The expression of staphylococcal enterotoxin genes was evaluated both qualitatively and quantitatively. In present study were used S. aureus strains from culture collections as well as those newly-isolated from raw milk samples. The obtained results indicate the various expression of the different genes for enterotoxin. However the main benefit of this work is the established protocol for the study of enterotoxin gene expression, which can provide a better understanding of the conditions for the enterotoxins production. Keywords: staphylococcal enterotoxin, gene expression, multiplex PCR, real-time PCR, mRNA analysis Journal: Czech Journal of Food Sciences Pages: SII56-SII65 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/208/2009-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/208/2009-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0008.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:208-2009-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Igor Hochel Author-Workplace-Name: Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Jiří Škvor Author-Workplace-Name: Department of Medical Biophysics, 1 st Medical Faculty, Charles University in Prague, Prague, Czech Republic Title: Characterisation of antibodies for the immunochemical detection of Enterobacter sakazakii Abstract: An indirect competitive enzyme immunoassay of Enterobacter sakazakii has been developed. The rabbit polyclonal antibodies to heat-labile or heat-stable antigen of the type strain E. sakazakii CNCTC 5739T were prepared for these purposes. The detection limits of enzyme immunoassays were within the range 0.6-14.4 × 105 cells/ml. Antibodies raised to heat-labile antigen were serotype-specific. Although they contain non-specific IgG fractions binding periplasmatic and cytosol proteins, the interactions of these immunoglobulins are not manifested under conditions of ELISA developed. Keywords: Enterobacter sakazakii, heat-labile antigen, IgG, ELISA, cross-reactivity Journal: Czech Journal of Food Sciences Pages: SII66-SII74 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/206/2009-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/206/2009-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0009.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:206-2009-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Eva ŠVIRÁKOVÁ Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Ivana SLOŽILOVÁ Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Petr TICHOVSKÝ Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Milada Plocková Author-Workplace-Name: Department of Dairy and Fat Technology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Title: Effect of Lactococcus sp. on the growth of Listeria sp. in the model UHT milk system Abstract: The work was aimed at the growth suppression of cultured listerias strains by cultured lactococci strains or commercial mesophilic cheese cultures during common cultivations in the model UHT milk system (0.5% w/w of milk fat content) at 30°C during 18 h aerobically. Milk was primarily fermented by lactococci at the level of 108 CFU/ml and secondarily contaminated by listerias at the level of 103 CFU/ml. The most intensive growth suppressions of both Listeria innocua (CCM 5884 or Ln-03) strains were caused by Lactococcus lactis subsp. lactis (LCC 416 or CHCC 2281) strains or DELVO-ADD® 100-X DSF cheese culture; the listerias growth reductions was from the level of 103 CFU/ml to 100 CFU/ml. Obtained results should be applied to dairy industry provided that HACCP, GHP and GMP systems must be observed. Keywords: Lactococcus, antilisterial activity, lactic acid, nisin, Listeria, UHT milk Journal: Czech Journal of Food Sciences Pages: SII8-SII11 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/672-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/672-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0010.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:672-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Irena Němečková Author-Workplace-Name: Dairy Research Institute Ltd., Prague, Czech Republic Author-Name: Marta Pechačová Author-Workplace-Name: MILCOM Inc., Prague, Czech Republic Author-Name: Petr Roubal Author-Workplace-Name: Dairy Research Institute Ltd., Prague, Czech Republic Title: Problems with detection of proteolytic microorganisms and their undesirable activities in milk Abstract: Occurrence of proteolytic enzymes in milk is often associated with technological problems and sensory, rheological and functional defects of final dairy products. Thus, the simple, cost-effective and available laboratory method for evaluation of undesirable proteolysis risk is needed. In our work we have tested cultivation plate methods and chemical methods (formol titration, ammonium reflectometric determination, the Kjeldahl method, the agar-well diffusion assay and spectrofotometry after cleavage of azo-casein) to choose the proper ones which can provide information on undesirable proteolytic changes especially in raw milk. Although the microbiological analyses cannot detect enzymes indigenous to milk, but only the quantity of producers of microbial enzymes, they seem to be the most acceptable, particularly usage of the Glucose-Trypton-Yeast Extract agar with 10% vol. of sterile milk added before pouring onto plates (incubation at 30°C for 72 h). The chemical methods are not sensitive enough to analyse the real milk samples. Keywords: proteolysis, spoilage risk assessment, cultivation method, simple chemical methods, raw milk, dairy products, Bacillus cereus Journal: Czech Journal of Food Sciences Pages: SII82-SII89 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/666-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/666-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0011.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:666-CJFS Template-Type: ReDIF-Article 1.0 Author-Name: Iva Lacmanová Author-Workplace-Name: Department of Biochemistry and Microbiology and Author-Name: Jarmila Pazlarová Author-Workplace-Name: Department of Biochemistry and Microbiology and Author-Name: Marta Kostelanská Author-Workplace-Name: Department of Food Analysis, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Author-Name: Jana Hajšlová Author-Workplace-Name: Department of Food Analysis, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague, Czech Republic Title: PCR-based identification of toxinogenic Fusarium species Abstract: Aim of this study was to develop sensitive PCR assay for mycotoxin producing Fusarium species. Strains Fusarium oxysporum 4199 and Fusarium culmorum 4044 were used as representatives of this group. Primers JB chosen to demonstrate the affiliation to genus Fusarium were derived from ITS region of rDNA. Gene from trichothecene pathway Tri4 was employed to design primers for toxin biosynthesis. Specificity of PCR based on JB primers was tested on DNA isolated from F. culmorum 4044, F. oxysporum 4199, Aspergillus oryzae 4002 and Mucor circinelloides 4018, Trichoderma sp. Both Fusarium species gave positive reaction, while the later ones did not react. Primers based on Tri4 highly specific sequences were giving positive reaction only with DNA from F. culmorum 4044 and F. oxysporum 4199. DNA isolated from six samples of contaminated wheat grains gave positive result on the presence of genus Fusarium and mycotoxines by optimised PCR protocol using JB and Tri4 primers. The results corresponded to LC/MS analysis that was established quantitatively in all samples to ascertain the amount and type of fusarious mycotoxines. Keywords: toxinogenic Fusarium, trichothecenes, PCR, JB primers, Tri4 primers Journal: Czech Journal of Food Sciences Pages: SII90-SII94 Volume: 27 Issue: SpecialIssue2 Year: 2009 DOI: 10.17221/634-CJFS File-URL: http://cjfs.agriculturejournals.cz/doi/10.17221/634-CJFS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjf-200911-0012.txt Handle: RePEc:caa:jnlcjf:v:27:y:2009:i:SpecialIssue2:id:634-CJFS