Klasična i molekularna identifikacija termotolerantnih kampilobaktera iz piletine

Poultry meat samples from Slovenian retail market were examined for the presence of thermotolerant campylobacters. The isolates were identified by phenotypic and genotypic methods. ISO 10272 recommendations were followed for phenotypic identification. Different PCR assays, targeting species specific DNA regions in C. jejuni and C. coli, were checked for their applicability in identification. High degree of tested samples was positive (27/33), with significant proportion of C. coli (32 %) among identified strains. High percentage of C. jejuni strains (54 %) were hippurate negative. Phenotypic identification was therefore found to be inconvenient because of the presence of the strains with atypical phenotype and possible misinterpretation of test results. Multiplex PCR, targeting hippuricase gene in C. jejuni and species specific region in C. coli, was found to be an efficient method that allowed fast, simple and accurate identification of C. jejuni and C. coli. FlaA PCR is a reliable method to identify the group C. jejuni/C. coli, but it does not differentiate between the two species. CdtB PCR is inconvenient because of many false negative and some false positive results.Ispitana je prisutnost termotolerantnih bakterija roda Campylobacter u uzorcima piletine sa slovenskog tržišta. Izolirani su sojevi identificirani fenotipskim i genotipskim postupcima. Fenotipska je identifikacija provedena klasičnim postupkom ISO 10272. Da bi se identificirali određeni sojevi, ispitana je primjenjivost različitih PCR postupaka, korištenjem specifičnih DNA regija za vrste C. jejuni i C. coli. Veliki broj ispitivanih uzoraka bio je pozitivan (27/33), sa značajnim udjelom C. coli u tim dvama identificiranim sojevima (32 %). Veliki udjel sojeva C. jejuni (54 %) bio je hipurat negativan. Fenotipska je identifikacija stoga neprikladan postupak zbog prisutnosti sojeva s atipičnim fenotipom, a i zbog mogućnosti pogrešne interpretacije rezultata testova. Multipleks PCR-postupak, koristeći hipurikazni gen u C. jejuni i u specifičnoj regiji za vrstu C. coli, pokazao se uspješnim jer omogućava brzu, jednostavnu i točnu identifikaciju C. jejuni i C. coli. Postupak PCR s lokusom flagelarnoga gena (flaA PCR) pouzdan je postupak za identifikaciju C. jejuni/C. coli, ali se njime ne mogu razlikovati te dvije vrste. Postupak PCR s lokusom gena citoletalnog toksina (cdtB PCR) nije prikladan jer daje puno lažno negativnih, a i nešto lažno pozitivnih rezultata

Klasična i molekularna identifikacija termotolerantnih kampilobaktera iz piletine

Poultry meat samples from Slovenian retail market were examined for the presence of thermotolerant campylobacters. The isolates were identified by phenotypic and genotypic methods. ISO 10272 recommendations were followed for phenotypic identification. Different PCR assays, targeting species specific DNA regions in C. jejuni and C. coli, were checked for their applicability in identification. High degree of tested samples was positive (27/33), with significant proportion of C. coli (32 %) among identified strains. High percentage of C. jejuni strains (54 %) were hippurate negative. Phenotypic identification was therefore found to be inconvenient because of the presence of the strains with atypical phenotype and possible misinterpretation of test results. Multiplex PCR, targeting hippuricase gene in C. jejuni and species specific region in C. coli, was found to be an efficient method that allowed fast, simple and accurate identification of C. jejuni and C. coli. FlaA PCR is a reliable method to identify the group C. jejuni/C. coli, but it does not differentiate between the two species. CdtB PCR is inconvenient because of many false negative and some false positive results.Ispitana je prisutnost termotolerantnih bakterija roda Campylobacter u uzorcima piletine sa slovenskog tržišta. Izolirani su sojevi identificirani fenotipskim i genotipskim postupcima. Fenotipska je identifikacija provedena klasičnim postupkom ISO 10272. Da bi se identificirali određeni sojevi, ispitana je primjenjivost različitih PCR postupaka, korištenjem specifičnih DNA regija za vrste C. jejuni i C. coli. Veliki broj ispitivanih uzoraka bio je pozitivan (27/33), sa značajnim udjelom C. coli u tim dvama identificiranim sojevima (32 %). Veliki udjel sojeva C. jejuni (54 %) bio je hipurat negativan. Fenotipska je identifikacija stoga neprikladan postupak zbog prisutnosti sojeva s atipičnim fenotipom, a i zbog mogućnosti pogrešne interpretacije rezultata testova. Multipleks PCR-postupak, koristeći hipurikazni gen u C. jejuni i u specifičnoj regiji za vrstu C. coli, pokazao se uspješnim jer omogućava brzu, jednostavnu i točnu identifikaciju C. jejuni i C. coli. Postupak PCR s lokusom flagelarnoga gena (flaA PCR) pouzdan je postupak za identifikaciju C. jejuni/C. coli, ali se njime ne mogu razlikovati te dvije vrste. Postupak PCR s lokusom gena citoletalnog toksina (cdtB PCR) nije prikladan jer daje puno lažno negativnih, a i nešto lažno pozitivnih rezultata

Effects of Low Temperature, Starvation and Oxidative Stress on the Physiology of Campylobacter jejuni Cells

Campylobacter jejuni is a food-borne pathogen that causes diarrheal diseases in humans and animals. Still unknown mechanisms allow the cells to overcome significant stresses despite the absence of several traditional stress response genes. C. jejuni is problematic because of its ability to adapt and survive in various conditions of extraintestinal environment, common to food production and supply chain. The authors demonstrated that preliminary exposure to 25 °C did not influence the heat resistance of C. jejuni cells at 55 °C. In addition, changes of morphology, culturability and viability were noticeable for fed and starved cultures exposed to low temperature and oxidative stresses. All physiology changes were influenced by starvation and, to a lesser extent, by other stresses. Starvation was the most significant factor in C. jejuni survival, coccoid cell formation and especially culturability. Temperature downshift, oxidative stress and starvation, usually in combination with other factors, can trigger transition of bacteria to a viable but nonculturable state. Results showed substantial variability in responses to stresses applied, indicating no correlation between low temperature and oxidative stress response of campylobacters. Understanding of microbial physiology after exposure to various stresses is essential for food safety

The Prevalence of Multiple Antibiotic Resistance in Campylobacter spp. From Retail Poultry Meat

Macrolides and fluoroquinolones are regarded as drugs of choice for the treatment of human Campylobacter infections. The use of antimicrobials for this purpose as well as in food animal production has resulted in the resistance of Campylobacter spp. to selected antibiotics. Since poultry is one of the most important sources of human Campylobacter infections the use of antibiotics in animal production can shorten the effective therapeutic life of antibiotics for human use. During 2001–2003, over 220 strains of C. jejuni and C. coli were isolated from 60 poultry meat samples from the retail market in Slovenia and further characterized by phenotypic and molecular methods. In this study, 55 sample-representative strains were tested for susceptibility to eight different antibiotics (ampicillin, amoxycillin/ clavulanic acid, ciprofloxacin, erythromycin, gentamicin, nalidixic acid, pefloxacin and tetracycline). Phenotypic procedures (disc diffusion test, E-test) as well as molecular detection of mutations (mismatch amplification mutation assay (MAMA) polymerase chain reaction (PCR)) in case of ciprofloxacin resistance were used. When assuming the results about antibiotic resistance, only 38.2 % of strains tested were susceptible to all antibiotics tested. Regarding ciprofloxacin, 58.2 % of tested strains were found to be resistant (minimal inhibitory concentration, MIC>4 mg/mL). The occurrence of resistance was much higher in C. coli (75.9 %) than in C. jejuni (38.5 %) isolates. The resistance rates to pefloxacin, nalidixic acid, erythromycin and tetracycline were 58.2, 49.1, 14.5 and 12.7 %, respectively. Eleven percent of strains were resistant to erythromycin and ciprofloxacin and 12.7 % of strains were resistant to tetracycline and quinolones. The results show the need for monitoring the prevalence and antibiotic resistance of zoonotic bacteria such as Campylobacter as well as the multiresistance phenomenon of Campylobacter isolates from food in our country

Pan-Cancer Analysis of lncRNA Regulation Supports Their Targeting of Cancer Genes in Each Tumor Context

Long noncoding RNAs (lncRNAs) are commonly dys-regulated in tumors, but only a handful are known toplay pathophysiological roles in cancer. We inferredlncRNAs that dysregulate cancer pathways, onco-genes, and tumor suppressors (cancer genes) bymodeling their effects on the activity of transcriptionfactors, RNA-binding proteins, and microRNAs in5,185 TCGA tumors and 1,019 ENCODE assays.Our predictions included hundreds of candidateonco- and tumor-suppressor lncRNAs (cancerlncRNAs) whose somatic alterations account for thedysregulation of dozens of cancer genes and path-ways in each of 14 tumor contexts. To demonstrateproof of concept, we showed that perturbations tar-geting OIP5-AS1 (an inferred tumor suppressor) andTUG1 and WT1-AS (inferred onco-lncRNAs) dysre-gulated cancer genes and altered proliferation ofbreast and gynecologic cancer cells. Our analysis in-dicates that, although most lncRNAs are dysregu-lated in a tumor-specific manner, some, includingOIP5-AS1, TUG1, NEAT1, MEG3, and TSIX, synergis-tically dysregulate cancer pathways in multiple tumorcontexts

Comparison of Slovenian and Estonian folk tales

In the first part of graduation thesis Comparison of slovenian and estonian folk tales a fairy tale is presented as a literary genre, along with its definition and types. The importance of fairy tale is explained according to various researchers in the field of folk tradition (Jack Zipes, Max Lüthi, Bruno Bettelheim, Alenka Goljevšček, etc.). In the following part children's literature is presented as a literary genre. Also, Slovenian and Estonian children's literature is explored. Finally, comparative folk-tale analysis (folklore theory, the structuralistic theory, literary theory, psychoanalytic theory, sociological theory, feminist theory, post-structural theory) are compared. The central part of the thesis contains a presentation of work by Hans-Jörg Uther and Vladimir Propp along with theoretical background. One of the notions in Uther’s theory is ATU index, acording to which fairy tales are selected. Propp’s theory includes the baseline situation, followed by presentation of thirty-one features characters. In the second part ATU types are presented. Each of the five types is illustrated with two folk tales – one Slovenianand one Estonian. The fifth slovenian fairy tale is authors fairy tale. Each of the stories is then analyzed according to Propp’s theory. Tales of individual types are compared with each other. The aim of the thesis is a comparative analysis meant to show the similarities and differences between folk tales. Despite different cultural and spatial influences, models of folk tales are similar with each other and yet, on the other hand, still different

Classical and Molecular Identification of Thermotolerant Campylobacters from Poultry Meat

Poultry meat samples from Slovenian retail market were examined for the presence of thermotolerant campylobacters. The isolates were identified by phenotypic and genotypic methods. ISO 10272 recommendations were followed for phenotypic identification. Different PCR assays, targeting species specific DNA regions in C. jejuni and C. coli, were checked for their applicability in identification. High degree of tested samples was positive (27/33), with significant proportion of C. coli (32 %) among identified strains. High percentage of C. jejuni strains (54 %) were hippurate negative. Phenotypic identification was therefore found to be inconvenient because of the presence of the strains with atypical phenotype and possible misinterpretation of test results. Multiplex PCR, targeting hippuricase gene in C. jejuni and species specific region in C. coli, was found to be an efficient method that allowed fast, simple and accurate identification of C. jejuni and C. coli. FlaA PCR is a reliable method to identify the group C. jejuni/C. coli, but it does not differentiate between the two species. CdtB PCR is inconvenient because of many false negative and some false positive results