Razvoj molekularnih metoda identifikacije psihoaktivnih gljiva i njihovih primjena u forenzici

doctoral thesis

• Title (english): Development of molecular methods for identification of psychoactive fungi and their implementation in forensic
• Author: Ivana Horjan
• Mentor: Višnja Besendorfer (mentor)
• Mentor: Zdenko Tkalčec (sumentor)
• Committee member: Zlatko Liber (predsjednik povjerenstva)
• Committee member: Domagoj Đikić (član povjerenstva)
• Committee member: Armin Mešić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2016, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology
• Universal decimal classification (UDC): 577 - Biochemistry. Molecular biology. Biophysics

Abstract

Većina psihoaktivnih vrsta gljiva sadrži psilocibin, psilocin, beocistin ili nor-beocistin, a zbog djelovanja psilocina sličnog LSD-u koriste se kao rekreacijska droga i zabranjene su. Dokazivanje aktivnih tvari iz forenzičkih uzoraka toksikološkim metodama GC-MS i LC-MS nije moguće iz otrusine spora ili micelija prije procesa fruktificiranja. Glavni cilj ovog rada bio je ukazati na potencijal primjene molekularnih i morfoloških metoda u forenzičkoj analizi psihoaktivnih gljiva. Morfološkom analizom 63 forenzička uzorka uspješno je identificirano 5 vrsta, a najzastupljenija je vrsta Psilocybe cubensis. Sekvenciranjem cijele regije ITS i dijela gena LSU forenzičkih uzoraka potvrđeni su rezultati morfološke analize i uspješno je identificirana šesta vrsta iz otrusine spora. Obje metode mogu se implementirati u rutinsku forenzičku analizu uzoraka gljiva. Potencijal istraživanog modela preliminarne identifikacije psihoaktivnih gljiva korištenjem specifičnih sondi TaqMan®MGB u reakciji Real Time PCR nije potvrđen. Analizom LC-MS otkriven je psihoaktivni status vrsta Stropharia semiglobata i Galerina badipes.

Abstract (english)

Most of psychoactive fungi contain psilocybin, psilocin, baeocystin or nor- baeocystin, and due to effects of psilocin, which are similar to LSD, they are illegal. Proving active substances from forensic samples through toxicological methods is not possible from spore print or from mycelia. Main goal of this thesis was to emphasize the potential of using molecular and morphological methods in forensic analysis of psychoactive fungi. Morphological analysis of 63 forensic samples resulted with 5 successfully identified species with Psilocybe cubensis as the most common. Through sequencing the ITS region and the part of LSU gene of forensic samples, morphological analysis results were confirmed and sixth species was successfully identified from spore print. Both methods can be implemented in the routine forensic workflow for fungi. Potential of the researched model for preliminary identification of psychoactive fungi by using specific TaqMan®MGB probes in RT-PCR reaction, was not confirmed. LC-MS analysis established the psychoactive status of species Stropharia semiglobata and Galerina badipes.

• Keywords: psihoaktivne gljive; morfologija; sekvenciranje DNA; sonde TaqMan®MGB; analiza LC-MS
• Keywords (english): sychoactive fungi; morphology; DNA sequencing; probes TaqMan®MGB; LC-MS analysis
• Language: croatian
• URN:NBN: urn:nbn:hr:217:183152
• Study programme: Title: Biology; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje prirodnih znanosti, polje biologija (doktor/doktorica znanosti, područje prirodnih znanosti, polje biologija)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Closed access
• Created on: 2017-05-12 10:39:33