20.500.12556/RUL-152590 Analiza viroma klopov vrste Ixodes ricinus v Sloveniji Analysis of the Ixodes ricinus tick virome in Slovenia Številne raziskave nakazujejo, da se v klopih vrste I. ricinus v Evropi nahajajo še neodkriti virusi. V magistrskem delu smo analizirali virom klopov I. ricinus v Sloveniji. Zanimala nas je struktura viroma RNA v različnih razvojnih stadijih klopov (larva, nimfa in odrasel klop) in na določenih zemljepisnih območjih. Zanimalo nas je, ali se v skupinah klopov nahajajo potencialno patogeni virusi. V raziskavo smo vključili 1056 klopov I. ricinus, ki smo jih vzorčili z metodo zastave, v 7 statističnih regijah Slovenije od meseca aprila do septembra 2022. Klope smo homogenizirali in združili v skupine. Osamili smo RNA in jo prepisali v dvoverižno cDNA po metodi SISPA. Pripravili smo knjižnice, knjižnice sekvencirali in izvedli bioinformatsko analizo, sestavljeno iz dveh stopenj. Večino odčitkov je se uvrstilo redovoma Bunyavirales in Reovirales. Soseske so se najpogosteje uvrstile družinam Nairoviridae, Phenuiviridae in Partitiviridae. Stopnja okužbe družin Nairoviridae, Phenuiviridae in Partitiviridae je bila najvišja pri odraslih klopih in se je med različnimi vzorčnimi mesti razlikovala. Odčitki so se v enem vzorcu kartirali na referenčni genom virusa Uukuniemi. Z odčitki in soseskami smo potrdili tudi virus klopnega meningoencefalitisa. Rezultati magistrskega dela kažejo, da so v viromu klopa I. ricinus v Sloveniji še neidentificirani virusi, ki jih moramo podrobneje raziskati, da ugotovimo ali lahko povzročijo bolezen pri človeku. Several studies indicate that I. ricinus ticks harbour undiscovered viruses in Europe. In this thesis, we aimed to analyse the virome of I. ricinus ticks in Slovenia. We were interested in the composition of the RNA virome, comparison between different developmental stages of ticks (larva, nymph and adult tick) and in specific geographical locations. We were interested in the presence of potentially pathogenic viruses in tick pools. We included 1056 I. ricinus ticks sampled by the flagging method in 7 statistical regions from April to September 2022. Ticks were homogenised and pooled. RNA was extracted and transcribed into ds cDNA by the SISPA method. Libraries were constructed, sequenced and a two-stage bioinformatic analysis was performed. Reads from each tick pool were classified within the orders Bunyavirales and Reovirales. Contigs were classified within the families Nairoviridae, Phenuiviridae and Partitiviridae. Infection rates within the families Nairoviridae, Phenuiviridae and Partitiviridae were the highest in adult ticks and varied between tick collection sites. Reads in one sample were mapped on the reference genome of the Uukuniemi virus. We also confirmed through reads and contigs presence of tick-borne meningoencephalitis virus. The results of the master's thesis indicate the presence of yet unidentified viruses in the I. ricinus tick virome in Slovenia. Further investigation is required to determine whether they could potentially cause diseases in humans. klopno prenosljivi virusi virom Ixodes ricinus metagenomika NGS tick-borne viruses virome Ixodes ricinus metagenomics NGS true false false [A. Pintar] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2023-11-30 07:15:16 2023-11-30 07:15:27 2023-12-04 07:43:58 0000-00-00 00:00:00 2023 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 579.61:595.421:575.111 235577 174631683 38192.pdf 38192.pdf 1 9449B4383358648A1ADF8F1C5744AD4C c5382442005f21bcf455922cf57009076fe03425d10ffbfba80933ad0799cf02 e03193a4-8f47-11ee-9a5d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=178150 Biotehniška fakulteta 0 0 0