Genska tipizacija arhiviranih uzoraka nedeterminiranih trakavica i njihovih larvalnih stadija iz domaćih i divljih životinja sekvenciranjem nad i cox gena

master's thesis

• Title (english): Gene Characterization by Nad and Cox Gene Sequence Analysis on the Undetermined Adult and Larval Cestodes Archived Samples from Domestic and Wild Animals
• Author: Monika Živković
• Mentor: Tatjana Živičnjak (mentor)
• Mentor: Relja Beck (komentor)
• Committee member: Dagny Stojčević Jan (predsjednik povjerenstva)
• Committee member: Tatjana Živičnjak (član povjerenstva)
• Committee member: Relja Beck (član povjerenstva)
• Granter: University of Zagreb; Faculty of Veterinary Medicine; (Department of Parasitology and Parasitic Diseases with Clinic); Zagreb
• Defense date and country: 2020-03-05, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Veterinary Medicine; Veterinary Clinical Sciences

Abstract

Morfološka identifikacija trakavica je složena i zahtjevna te podrazumijeva sve dobro očuvane dijelove za pravilnu determinaciju. Osnovni cilj je ovim diplomskim radom bilo genetičkim tipiziranjem odrediti vrstu trakavice ili larvalnog stadija iz arhiviranih uzoraka determiniranih nakon razudbe isključivo na osnovi morfologije.
Metodi lančane reakcije polimerazom sa ciljem genske tipizacije i određivanja vrste su podvrgnute trakavice ili larvalni oblici trakavica pronađeni kod 47 životinja. Izdvojena DNK je pretražena lančanom reakcijom polimerazom usmjerenom na umnažanje NAD i COX gena. Umnoženi odsječci su pročišćeni, sekvencionirani u oba smjera, te uspoređeni sa dostupnim sekvencama u banci gena GenBank®.
Molekularnim je metodama dokazano 10 vrsta trakavica. Po prvi puta u Hrvatskoj su na taj način dokazane vrste Moniezia expansa, Dipylidium caninum, Mesocestoides litteratus, Taenia arctos, Taenia cracciceps, Taenia hydatigena, Taenia krabbei, Taenia martis, Hydatigera taeniaeformis sensu stricto i genotip Echinococcus canadensis. U nositelja i posrednika dokazane su vrste Taenia krabbei i Taenia martis. Vrste Moniezia expansa, Dipylidium caninum, Mesocestoides litteratus, Tenia arctos, Taenia crassiceps i Hydatigera taeniaeformis sensu stricto su determinirane u nositelju, a vrste Taenia hydatigena i genotip Echinococcus canadensis u posredniku. Za 6 novih sekvenci su potrebna dodatna istraživanja.

Abstract (english)

The morphological identification of cestodes is complex and demanding and it requires well-preserved parasite for proper determination. The aim of the research was the molecular gene characterization of archived 47 samples of adult or larval cestodes that had been determined solely based on morphology after being found in animals during necropsy. Samples were subjected to DNA extractions, enzymatic amplifications and sequencing. The mtDNA regions were amplified using previously published NAD and COX primers. The replicated sections were purified, sequenced in both directions and compared with the available sequences in the GenBank®.
Finaly, 10 tapeworm species were molecularly detected. For the first time, Moniezia expansa, Dipylidium caninum, Mesocestoides litteratus, Taenia arctos, Taenia crassiceps, Taenia hydatigena, Taenia krabbei, Taenia martis, Hydatigera taeniaeformis s.s. and Echinococcus canadensis genotype were molecularly determined in Croatia. Taenia krabbei and Taenia martis were determined in definitive and intermediate host. Moniezia expansa, Dipylidium caninum, Mesocestoides litteratus, Tenia arctos, Taenia crassiceps and Hydatigera taeniaeformis s.s. were determined in the definitive host, and Taenia hydatigena and Echinococcus canadensis genotype in the intermediate host. Six new sequences showed less than 90% compatibility and require additional research.

• Keywords: Cestoda; PCR; NAD; COX; sekvenciranje
• Keywords (english): Cestoda; PCR; NAD; COX; sequencing
• Language: croatian
• URN:NBN: urn:nbn:hr:178:710707
• Study programme: Title: Veterinary Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica veterinarske medicine (doktor/doktorica veterinarske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-10-15 09:24:38