Analiza genetičke raznolikosti gljivice Malassezia pachydermatis izolirane iz smeđeg medvjeda (Ursus arctos) na području Republike Hrvatske

master's thesis

• Title (english): A Genetic Diversity Analysis of the Brown Bear (Ursus Arctos) Malassezia Pachydermatis Yeast Isolates on the Territory of the Republic of Croatia
• Author: Ivana Ivanković
• Mentor: Suzana Hađina (mentor)
• Committee member: Zoran Milas (predsjednik povjerenstva)
• Committee member: Ljiljana Pinter (član povjerenstva)
• Committee member: Suzana Hađina (član povjerenstva)
• Granter: University of Zagreb; Faculty of Veterinary Medicine; (Department of Microbiology and Infectious Diseases with Clinic); Zagreb
• Defense date and country: 2018-01-19, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Veterinary Medicine

Abstract

Smeđi medvjed (Ursus arctos) najveća je kopnena zvijer koja nastanjuje područja Gorskog kotara i Like te je uvršten na popis ugroženih biljnih i životinjskih vrsta Hrvatske. Gljivica Malassezia pachydermatis, koja se smatra oportunističkim patogenom, uslijed određenih predisponirajućih čimbenika često uzrokuje različite patološke promjene na koži kućnih ljubimaca, ali i domaćih i divljih životinja. Do danas postoji vrlo malo podataka o prisutnosti i ulozi gljivice M. pachydermatis na koži medvjeda. Opći cilj ovog istraživanja bio je izolirati genomsku DNK i optimizirati izolaciju genomske DNK iz kultura M. pachydermatis naraslih na dvije različite hranjive podloge. Specifičan cilj bio je umnožavanje specifičnog odsječka DNK uz pomoć početnica za dva genetička biljega, ITS1 i LSU, u svrhu ustanovljavanja genetičke raznolikosti gljivice M. pachydermatis u smeđeg medvjeda (Ursus arctos) na području Republike Hrvatske. Genomska DNK izolirana je iz narasle kulture gljivice M. pachydermatis komercijalno dostupnim kitom, specifični odsječci DNK (ITS1 i LSU regija) umnoženi su lančanom reakcijom polimeraze te je analiziran slijed nukleotida. Na osnovu dobivenih rezultata ustanovljeno da umnoženi odsječci DNK svojom veličinom odgovaraju objavljenim veličinama ITS1 i LSU regije. Osim toga uočeno je da u analiziranim regijama postoje određene specifičnosti u slijedu nukleotida te da je LSU regija u odnosu na ITS1 znatno bolje očuvana. Navedeni preliminarni rezultati ukazuju na potencijalnu evolucijsku očuvanost koju je potrebno potvrditi analizom većeg broja uzoraka i uključivanjem dodatnih molekularnih metoda.

Abstract (english)

The brown bear (Ursus arctos) is the largest terrestrial carnivore that inhabits the Mountain District of Gorski kotar and the geographical area of Lika. It is included in the red list of the endangered species in Croatia. Malassezia pachydermatis yeast, which is considered an opportunistic pathogen, causes various pathological skin disorders in pets under certain predisposing conditions, as well as in domestic and wild animals. There is very little data about the presence and role of M. pachydermatis on the bear skin. The first aim of the study was to optimize a genomic DNA extraction protocol from the M. pachydermatis culture growing on two different culture media. The specific aim was the amplification of particular DNA fragments using ITS1 and LSU primer sequences in order to describe the genetic variation of Malassezia pachydermatis yeast in the brown bear (Ursus arctos) on the territory of Croatia. The frozen archived yeast strains were grown on two different culture media, the genomic DNA was isolated using a commercially available kit and the specific DNA fragments (ITS1 and LSU regions) were amplified by a polymerase chain reaction. In addition, the nucleotide sequences were analyzed. The results have shown that the isolated yeast colonize both the epithelium of the ear canal and the perianal area of the brown bear. The multiplied DNA fragments correspond to the previously published length of the ITS1 and LSU regions. It has been noticed that the analyzed regions contain certain varieties of the nucleotide sequence and that the LSU region is much better preserved than the ITS1 region. These preliminary results indicate possible evolutionary preservation, which needs to be further explored by analyzing a larger number of samples and by introducing additional molecular research methods.

• Keywords: genetička raznolikost; Malassezia pachydermatis; smeđi medvjed; Republika Hrvatska
• Keywords (english): genetic diversity; Malassezia pachydermatis; brown bear; Republic of Croatia
• Language: croatian
• URN:NBN: urn:nbn:hr:178:173283
• 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
• Repository: Repository of Faculty of Veterinary Medicine
• Created on: 2018-07-25 07:26:38