Ugotavljanje in molekularna opredelitev virusov ORF pri gamsu (Rupicapra rupicapra) ter papilomavirusov pri srni (Capreolus capreolus) in navadnem jelenu (Cervus elaphus)
| Autor: | Kmetec, Jernej |
|---|---|
| Přispěvatelé: | Vengušt, Gorazd |
| Jazyk: | slovinština |
| Rok vydání: | 2021 |
| Předmět: |
contagious
sekvenciranje celotnega genoma – metode orf virus – genetika papilomavirusne okužbe – virologija – patologija orf virus – genetics Deer – virology Divjad – virologija phylogeny papillomaviridae – genetics – classification viral – blood virusni genom papilomaviridae – genetika – klasifikacija filogenija antibodies ecthyma papillomavirus infection – virology – pathology protitelesa proti virusom – kri genome whole genome sequencing – methods kužni ektim viral |
| Popis: | Kužni ektim povzroča virus Orf, ki spada v družino Poxviridae, rod Parapoxvirus. V doktorski nalogi opisujemo analizo dveh genomov izolatov virusov Orf pri gamsih (sev ORFV_82/13 in sev ORFV_12/17). Z metodo sekvenciranja naslednje generacije (angl. next-generation sequencing, NGS) smo prvi določili skoraj celotno nukleotidno zaporedje genomov (angl. nearly complete genome) virusov Orf pri gamsih. Analiza genomov virusov Orf pri gamsih je pokazala visoko stopnjo sorodnosti z virusi Orf pri ovcah in kozah. Genoma sta na nivoju nukleotidnega zaporedja med seboj 99,93-odstotno podobna in se od genomov drugih virusov Orf, dostopnih v podatkovni zbirki NCBI GenBank, razlikujeta od 7,61 % (ovčji sev NZ2) do 11,62 % (kozji sev NP). Filogenetska analiza genov osrednje regije je pokazala, da sta virusa Orf pri gamsih tvorila svojo filogenetsko podskupino. V doktorski nalogi smo z ugotavljanjem prisotnosti protiteles proti virusom Orf s serum nevtralizacijskim testom ugotavljali prekuženost populacije gamsov z virusom Orf. Testirali smo 40 vzorcev serumov gamsov. Rezultati testa so bili negativni pri 38 vzorcih, dveh rezultatov ni bilo mogoče odčitati zaradi citotoksičnega učinka vzorca seruma na celično kulturo. S pregledom arhivskih vzorcev zadnjih dveh desetletij, upoštevajoč posredovane podatke lovskih družin ter izkušnje Inštituta za patologijo, divjad, ribe in čebele, smo ugotovili ciklično pojavljanje kužnega ektima pri gamsih. Fibropapilomi so benigni tumorji kože, ki jih povzročajo virusi iz družine Papillomaviridae. Bolezen z izraženimi kliničnimi znaki kaže v Evropi tendenco širjenja med populacijo srn, medtem ko je pri navadnem jelenu opažena redko. V nalogi opisujemo primere fibropapilomatoze pri srnah in navadnem jelenu z značilno klinično in histopatološko sliko. Z metodo NGS smo določili nukleotidno zaporedje celotnih genomov sedmih papilomavirusov (PV), in sicer šestim sevom PV pri srnah (CcaPV1 sev 31-16, CcaPV1 sev 32-16, CcaPV1 sev 37-14, CcaPV1 sev 63-15, CcaPV1 sev 103-16 in CcaPV1 sev 84-14) in enemu sevu PV pri navadnem jelenu (CePV1v sev 64-14). Pri srnah smo potrdili okužbo s tipom CcaPV1 in pri navadnem jelenu s podtipom CePV1v. Primerjava virusnih genomov z že znanimi genomi iz NCBI GenBank je pokazala visoko genetsko stabilnost genoma v času in prostoru. S pregledom arhivskih vzorcev zadnjih dveh desetletij smo ugotovili pojavljanje fibropapilomatoze pri srnah na letni ravni, pri navadnem jelenu pa izredno redko. Contagious ectyma is a disease caused by Orf virus, a member of the family Poxviridae, genus Parapoxvirus. In the doctoral dissertation, we describe the analysis of two genomes of Orf virus isolates in chamois (strain ORFV_82/13 and strain ORFV_12/17). Using next-generation sequencing (NGS), we were the first ones to identify nearly complete genome of Orf viruses in chamois. The analyses revealed a high degree of similarity to Orf virus found in sheep and goats. At the nucleotide sequence level, the genomes are 99.93 % identical to each other and differ from other Orf viruses from 7.61 % (sheep strain NZ2) to 11.62 % (goat strain NP). Phylogenetic analysis of genes in the central region has shown that chamois Orf viruses form a distinct phylogenetic group. In the doctoral dissertation, the presence of antibodies against Orf viruses by serum neutralization assay was used to determine the seroprevalence of chamois population with Orf virus. Forty serum samples from chamois were tested. Test results were negative for 38 samples, and two results could not be read due to the cytotoxic effect of the serum sample on the cell culture. By reviewing the archival samples of the last two decades, considering the data provided by hunting organisations and the experience of the Institute of Pathology, wild animals, fish and bees, we found cyclic occurrence of contagious ecthyma in chamois. Fibropapillomas are benign skin tumours caused by viruses of the Papillomaviridae family. The disease with pronounced clinical signs shows a tendency to spread among the roe deer population in Europe, whereas it is rarely observed in red deer. The dissertation describes cases of fibropapillomatosis in roe deer and red deer with a characteristic clinical and histopathological picture. NGS method was used to determine the nucleotide sequence of the whole genome of seven papillomaviruses (PV), namely the sixth PV strain in roe deer (CcaPV1 strain 31-16, CcaPV1 strain 32-16, CcaPV1 strain 37-14, CcaPV1 strain 63-15, CcaPV1 strain 103-16 and CcaPV1 strain 84-14) and one PV strain in red deer (CePV1v strain 64-14). Infections with the CcaPV1 type were confirmed in roe deer and with the CePV1v subtype in red deer. Comparison of the viral genomes with previously known genomes from NCBI GenBank showed high genetic stability of the genome in time and space. Reviewing archival samples from the last two decades, it was found that fibropapillomatosis occurs annually in roe deer but extremely rarely in red deer. |
| Databáze: | OpenAIRE |
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