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Papillomaviruses (PVs) are an extremely large group of viruses that cause skin and mucosa infections in humans and various animals. In roe deer and red deer, most PVs belong to the Deltapapillomavirus genus and cause neoplastic changes that are generally described as fibropapillomas. Despite the wide distribution of roe and red deer throughout Europe and beyond, the data in the scientific literature regarding the widespread distribution of PVs and the genetic variability of PV genomes in these species are rather scarce. This study describes cutaneous fibropapillomatosis cases in roe and red deer with clinical manifestations that are typical of infections with PVs. In all cases, the presence of PV DNA was confirmed using PCR, followed by Sanger sequencing of the partial L1 gene. The complete PV genomes were determined in all the investigated samples using next-generation sequencing technology, revealing infections of roe deer with the CcaPV1-type and red deer with the CePV1v-type variant. A comparison of the complete CcaPV1-type and CePV1v-type variant genome sequences reported here with already available complete genome sequences in GenBank revealed their great genetic stability across time and space.

The aim of this research was to investigate use of an indirect enzyme-linked immunosorbent assay (ELISA) and Western blotting test with native excretory/secretory Fascioloides magna antigen (ES-Ag) in free-ranging red deer (Cervus elaphus). The research was performed on 48 red deer shot during routine culls from two different areas considering occurrence of fascioloidosis. After the coprological examination and gross pathology examination of the livers for F. magna, serum samples were divided into three groups as infected (n=32), uninfected (n=13) and previously infected deer (n=3). Indirect ELISA results were significantly higher for serum samples from infected deer (percentage of positivity (PP) 65.1±18.4) than uninfected (PP 11.6±13.7) and previously infected deer (PP 20.3±6.4). Samples from uninfected had fewer bands (30-33 and 104 kDa) on Western blotting than samples from infected deer (6, 17, 22, 27, 30-33, 40, 45 and 104 kDa). The number of alive flukes were positively correlated with ELISA results and the number of F. magna eggs per gram of faeces (EPG). The results indicated that ELISA and Western blotting test could be useful in the early diagnosis of fascioloidosis in red deer and in the control in a population of free-ranging red deer as well.

Background: In two last decades, Fascioloides magna caused significant production losses in open hunting grounds in several endemic areas in Croatia. In order to improve diagnostic measures we investigated the effects of Fascioloides magna infection on the serum biochemistry values in naturally infected Red deer population in the northern Croatia. Methods: Blood samples from 22 Fascioloides magna infected deer and from 19 non-infected were taken from the heart immediately after they were shot. The liver from each animal was sliced into 1.0 to 2.0 cm wide, parallel slices, and carefully examined. Fecal samples were collected from each deer, and fecal examination was performed using the sedimentation method. Results: Significantly higher values for aspartate aminotransferase (AST), lactate dehydrogenase (LDH), glutamate dehydrogenase (GDH) and globulin were recorded in Fascioloides magna infected deer, whereas glucose value and albumine/globuline ratio were significantly higher in deer without Fascioloides magna. Conclusions: It has been concluded that serum biochemistry could be used in diagnosis of Fascioloides magna infections in Red deer population.

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.