20.500.12556/DKUM-42814 IZOLACIJA BAKTERIJ IZ RODU Clostridium IZ VZORCEV PRSTI Isolation of bacteria from genus Clostridium from soil samples Klostridiji so paličaste, obligatno anaerobne, po Gramu pozitivne bakterije, ki tvorijo izjemno odporne spore. Zaradi slednjega so klostridiji ubikvitarni, saj jih lahko najdemo v okoljih kot so prst, okoljske vode, prisotni pa so tudi v človeškem in živalskem prebavilu. Ena izmed pomembnih vrst tega rodu je bakterija Clostridium difficile, ki je trenutno v svetu eden izmed pomembnejših povzročiteljev bolnišničnih okužb. V zadnjem času pa narašča tudi število izvenbolnišničnih okužb. Bolezen povzročajo le toksigeni sevi, ki izdelujejo toksin A in/ali toksin B, ki sta glavna dejavnika virulence C. difficile. V diplomskem delu smo iz 6 različnih vrst/tipov prsti osamili več klostridijskih vrst, tudi C. difficile. Izolate smo identificirali s komercialnim sistemom MALDI Biotyper (Bruker Daltonics), ki je komercialna fenotipska metoda za identifikacijo in temelji na analizi proteinov s pomočjo laserske svetlobe. Izolate C. difficile smo še karakterizirali, in sicer z dvema dobro razširjenima genotipizacijskima metodama: toksinotipizacijo in PCR ribotipizacijo. Skupno smo osamili 34 klostridijskih izolatov, ki so pripadali 7 različnim vrstam. Med njimi so prevladovale vrste Clostridium glycolicum (n=11), Clostridium bifermentans (n=5) in Clostridium mayombei (n=4). C. difficile (n=10) smo zaznali le v vzorcu sedimenta reke Drave, s selektivno gojitveno metodo. Skupno deset izolatov iz tega vzorca je bilo netoksigenih in PCR ribotipa 010. Največje število in raznolikost klostridijev smo določili v vzorcu komposta. Z raziskavo v diplomskem delu smo potrdili, da lahko v različnih okoljih pričakujemo različne predstavnike rodu Clostridium, raznolikost vrst pa je odvisna od tipa okolja nabranih vzorcev. Bakterije C. difficile sicer nismo našli v različnih vzorcih prsti, pač pa v rečnem sedimentu, kjer do sedaj v Sloveniji še ni bil opisan. Clostridia are obligatory anaerobic, Gram-positive rods, which form very resistant spores. The latter makes them ubiquitous and could therefore be detected in soil, surface waters, and are also present in human and animal gut. One of the important species from the genus Clostridium is Clostridium difficile, now considered to be one of very important nosocomial pathogen. However, in recent years a significant increase in number of community-acquired C. difficile infections was observed. Disease is caused only by toxigenic strains that produce toxin A and/or toxin B, which are the main C. difficile virulence factors. In Graduation Thesis different Clostridia species, including C. difficile were isolated from 6 different soil samples. Isolates have been identified by commercial MALDI Biotyper system (Bruker Daltonics), which is phenotypic identification method, based on the protein analysis using laser light. Furthermore, C. difficile isolates have been characterized by two widely used genotyping methods: toxinotyping and PCR ribotyping. Altogether 34 clostridia isolates were obtained and belonged to 7 different species, where Clostridium glycolicum (n=11), Clostridium bifermentans (n=5) and Clostridium mayombei (n=4) predominated. C. difficile (n=10) was isolated only from one sample; sediment of river Drava by selective cultivation method. All 10 isolates characterized as non-toxigenic strain of PCR ribotype 010. In compost sample the highest number and strains diversity of isolated clostridia was determined. Within this research, multiple Clostridium species from soil were determined. Diversity and the number depended on the soil type. We didn't find C. difficile in any of the tested soil samples, however, it was detected in the river sediment, where up to now in Slovenia has not yet been described. klostridiji prst Clostridium difficile identifikacija toksinotipizacija PCR ribotipizacija raznolikost vrst Clostridia soil Clostridium difficile identification toxinotyping PCR ribotyping species diversity true true false [T. Rikanović] Slovenski jezik Angleški jezik Diplomsko delo 2013-10-20 16:56:22 2013-11-13 13:13:55 2022-05-08 03:06:04 2021-04-27 03:39:24 2013 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 579.852.13(043.2) 20168968 URN:SI:UM:DK:UVIYBD0R UNI_Rikanovic_Tanja_2013.pdf UNI_Rikanovic_Tanja_2013.pdf 1 D72F1E8DD7B915512F9BFF4E6759E9C8 41ba18ca273c59c209c62287a289057f20fe1ddbb6a4423bea83859e9ea442dd 7e7de655-7c0a-11eb-bb7a-00155d0001ca https://dk.um.si/Dokument.php?lang=slv&id=60231 Fakulteta za naravoslovje in matematiko 0 0 0