State-of-the-Art Animal Viruses Research in Slovenia

A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Animal Viruses".

Deadline for manuscript submissions: closed (31 March 2021) | Viewed by 25312

Special Issue Editor


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Guest Editor
1. Virology Unit, Institute of microbiology and parasitology, Gerbičeva ulica 60, SI-1000 Ljubljana, Slovenia
2. National Veterinary Institute, Veterinary Faculty, University of Ljubljana, Gerbičeva ulica 60, SI-1000 Ljubljana, Slovenia
Interests: virology; molecular epidemiology; diagnostics; transmission; sequencing

Special Issue Information

Dear Colleagues,

Slovenia is one of the smallest European countries, but very dynamic and with intensive trade and research in animal viruses. Along with the detection of emerging and re-emerging viral diseases, control programmes have been successfully in the running to eradicate several diseases. Despite great success over the last few decades, several viral pathogens remain challenging. Molecular epidemiology studies and novel technologies, such as next generation sequencing, provide good opportunities for better understanding special features of viral pathogens, transmission of viruses within and between countries, and development of new viral disease control strategies.

In this Special Issue, we will focus on detection, prevalence, control programmes, molecular epidemiology studies, transmission, eradication of endemically present, emerging, and re-emerging animal viruses in Slovenia.

Dr. Ivan Toplak
Guest Editor

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Keywords

  • emerging and re-emerging viruses
  • detection
  • prevalence
  • control programmes
  • molecular epidemiology
  • transmission
  • eradication

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Published Papers (8 papers)

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Research

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12 pages, 1488 KiB  
Article
First Report of Phodopus sungorus Papillomavirus Type 1 Infection in Roborovski Hamsters (Phodopus roborovskii)
by Grega Gimpelj Domjanič, Lea Hošnjak, Maja M. Lunar, Lucijan Skubic, Tomaž Mark Zorec, Joško Račnik, Blaž Cigler and Mario Poljak
Viruses 2021, 13(5), 739; https://doi.org/10.3390/v13050739 - 23 Apr 2021
Cited by 3 | Viewed by 2823
Abstract
Papillomaviruses (PVs) are considered highly species-specific with cospeciation as the main driving force in their evolution. However, a recent increase in the available PV genome sequences has revealed inconsistencies in virus–host phylogenies, which could be explained by adaptive radiation, recombination, host-switching events and [...] Read more.
Papillomaviruses (PVs) are considered highly species-specific with cospeciation as the main driving force in their evolution. However, a recent increase in the available PV genome sequences has revealed inconsistencies in virus–host phylogenies, which could be explained by adaptive radiation, recombination, host-switching events and a broad PV host range. Unfortunately, with a relatively low number of animal PVs characterized, understanding these incongruities remains elusive. To improve knowledge of biology and the spread of animal PV, we collected 60 swabs of the anogenital and head and neck regions from a healthy colony of 30 Roborovski hamsters (Phodopus roborovskii) and detected PVs in 44/60 (73.3%) hamster samples. This is the first report of PV infection in Roborovski hamsters. Moreover, Phodopus sungorus papillomavirus type 1 (PsuPV1), previously characterized in Siberian hamsters (Phodopus sungorus), was the only PV detected in Roborovski hamsters. In addition, after a detailed literature search, review and summary of published evidence and construction of a tanglegram linking the cladograms of PVs and their hosts, our findings were discussed in the context of available knowledge on PVs described in at least two different host species. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
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10 pages, 1323 KiB  
Article
Genetic Characterisation and Comparison of Three Human Coronaviruses (HKU1, OC43, 229E) from Patients and Bovine Coronavirus (BCoV) from Cattle with Respiratory Disease in Slovenia
by Monika Jevšnik Virant, Danijela Černe, Miroslav Petrovec, Tomislav Paller and Ivan Toplak
Viruses 2021, 13(4), 676; https://doi.org/10.3390/v13040676 - 15 Apr 2021
Cited by 5 | Viewed by 2838
Abstract
Coronaviruses (CoV) are widely distributed pathogens of human and animals and can cause mild or severe respiratory and gastrointestinal disease. Antigenic and genetic similarity of some CoVs within the Betacoronavirus genus is evident. Therefore, for the first time in Slovenia, we investigated the [...] Read more.
Coronaviruses (CoV) are widely distributed pathogens of human and animals and can cause mild or severe respiratory and gastrointestinal disease. Antigenic and genetic similarity of some CoVs within the Betacoronavirus genus is evident. Therefore, for the first time in Slovenia, we investigated the genetic diversity of partial 390-nucleotides of RNA-dependent-RNA polymerase gene (RdRp) for 66 human (HCoV) and 24 bovine CoV (BCoV) positive samples, collected between 2010 and 2016 from human patients and cattle with respiratory disease. The characterized CoV strains belong to four different clusters, in three separate human clusters HCoV-HKU1 (n = 34), HCoV-OC43 (n = 31) and HCoV 229E (n = 1) and bovine grouping only as BCoVs (n = 24). BCoVs from cattle and HCoV-OC43 were genetically the most closely related and share 96.4–97.1% nucleotide and 96.9–98.5% amino acid identity. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
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19 pages, 2441 KiB  
Article
Molecular and Phylogenetic Characterization of Novel Papillomaviruses Isolated from Oral and Anogenital Neoplasms of Japanese Macaques (Macaca fuscata)
by Lucijan Skubic, Lea Hošnjak, Jeannette P. Staheli, Michael R. Dyen, Rebecca M. Ducore, Lois M. A. Colgin, Anne D. Lewis and Mario Poljak
Viruses 2021, 13(4), 630; https://doi.org/10.3390/v13040630 - 7 Apr 2021
Viewed by 3444
Abstract
Papillomaviruses (PVs) are a diverse group of host species-specific DNA viruses, etiologically linked with various benign and malignant neoplasms of cutaneous and mucosal epithelia. Here, we describe the detection and characterization of the first two PVs naturally infecting Japanese macaques (Macaca fuscata [...] Read more.
Papillomaviruses (PVs) are a diverse group of host species-specific DNA viruses, etiologically linked with various benign and malignant neoplasms of cutaneous and mucosal epithelia. Here, we describe the detection and characterization of the first two PVs naturally infecting Japanese macaques (Macaca fuscata), including the determination of their etiological association(s) with the development of original neoplasms. The molecular and phylogenetic analyses were performed on complete genome sequences of Macaca fuscata PV types 1 (MfuPV1) and 2 (MfuPV2), which were completely sequenced in samples of a malignant oral tumor and benign anogenital neoplasm of Japanese macaques, respectively. Subsequently, two type-specific quantitative real-time PCRs were developed to estimate viral loads of MfuPV1 and MfuPV2 and to evaluate their etiological roles. The in silico molecular analyses revealed that both viral genomes encode characteristic PV proteins with conserved functional domains and have a non-coding genomic region with regulatory sequences to regulate and complete the viral life cycle. However, additional experimental evidence is needed to finally confirm the presence and biological functionality of the molecular features of both novel PVs. While MfuPV1, together with PVs identified in other macaques, is classified into the Alphapapillomavirus (Alpha-PV) species 12, MfuPV2 is most likely a representative of the novel viral species within the Alpha-PV genus. Their relatively high viral loads suggest that both PVs are etiologically linked with the development of the original neoplasms. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
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9 pages, 252 KiB  
Article
A Sentinel Serological Study in Selected Zoo Animals to Assess Early Detection of West Nile and Usutu Virus Circulation in Slovenia
by Pavel Kvapil, Joško Račnik, Marjan Kastelic, Eva Bártová, Miša Korva, Mateja Jelovšek and Tatjana Avšič-Županc
Viruses 2021, 13(4), 626; https://doi.org/10.3390/v13040626 - 6 Apr 2021
Cited by 7 | Viewed by 2496
Abstract
Monitoring infectious diseases is a crucial part of preventive veterinary medicine in zoological collections. This zoo environment contains a great variety of animal species that are in contact with wildlife species as a potential source of infectious diseases. Wild birds may be a [...] Read more.
Monitoring infectious diseases is a crucial part of preventive veterinary medicine in zoological collections. This zoo environment contains a great variety of animal species that are in contact with wildlife species as a potential source of infectious diseases. Wild birds may be a source of West Nile virus (WNV) and Usutu (USUV) virus, which are both emerging pathogens of rising concern. The aim of this study was to use zoo animals as sentinels for the early detection of WNV and USUV in Slovenia. In total, 501 sera from 261 animals of 84 animal species (including birds, rodents, lagomorphs, carnivores, ungulates, reptiles, equids, and primates) collected for 17 years (2002–2018) were tested for antibodies to WNV and USUV. Antibodies to WNV were detected by indirect immunofluorescence tests in 16 (6.1%) of 261 animals representing 10 species, which were sampled prior to the first active cases of WNV described in 2018 in Slovenia in humans, a horse, and a hooded crow (Corvus cornix). Antibodies to USUV were detected in 14 out of 261 animals tested (5.4%) that were positive prior to the first positive cases of USUV infection in common blackbirds (Turdus merula) in Slovenia. The study illustrates the value of zoological collections as a predictor of future emerging diseases. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
16 pages, 1366 KiB  
Article
Determination of Genetically Identical Strains of Four Honeybee Viruses in Bumblebee Positive Samples
by Ivan Toplak, Laura Šimenc, Metka Pislak Ocepek and Danilo Bevk
Viruses 2020, 12(11), 1310; https://doi.org/10.3390/v12111310 - 16 Nov 2020
Cited by 11 | Viewed by 2633
Abstract
In recent years, there has been growing evidence that certain types of honeybee viruses could be transmitted between different pollinators. Within a voluntary monitoring programme, 180 honeybee samples (Apis mellifera carnica) were collected from affected apiaries between 2007 and 2018. Also [...] Read more.
In recent years, there has been growing evidence that certain types of honeybee viruses could be transmitted between different pollinators. Within a voluntary monitoring programme, 180 honeybee samples (Apis mellifera carnica) were collected from affected apiaries between 2007 and 2018. Also from August 2017 to August 2018, a total 148 samples of healthy bumblebees (Bombus lapidarius, B. pascuorum, B. terrestris, B. lucorum, B. hortorum, B. sylvarum, B. humilis) were collected at four different locations in Slovenia, and all samples were tested by using RT-PCR methods for six honeybee viruses. Direct sequencing of a total 158 positive samples (acute bee paralysis virus (ABPV n = 33), black queen cell virus (BQCV n = 75), sacbrood bee virus (SBV n = 25) and Lake Sinai virus (LSV n = 25)) was performed from obtained RT-PCR products. The genetic comparison of identified positive samples of bumblebees and detected honeybee field strains of ABPV, BQCV, SBV, and LSV demonstrated 98.74% to 100% nucleotide identity between both species. This study not only provides evidence that honeybees and bumblebees are infected with genetically identical or closely related viral strains of four endemically present honeybee viruses but also detected a high diversity of circulating strains in bumblebees, similar as was observed among honeybees. Important new genetic data for endemic strains circulating in honeybees and bumblebees in Slovenia are presented. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
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14 pages, 2379 KiB  
Article
A Comprehensive Study of Cutaneous Fibropapillomatosis in Free-Ranging Roe Deer (Capreolus capreolus) and Red Deer (Cervus elaphus): from Clinical Manifestations to Whole-Genome Sequencing of Papillomaviruses
by Jernej Kmetec, Urška Kuhar, Aleksandra Grilc Fajfar, Diana Žele Vengušt and Gorazd Vengušt
Viruses 2020, 12(9), 1001; https://doi.org/10.3390/v12091001 - 8 Sep 2020
Cited by 4 | Viewed by 3064
Abstract
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. [...] Read more.
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. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
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Review

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14 pages, 2627 KiB  
Review
The Successful Elimination of Sylvatic Rabies Using Oral Vaccination of Foxes in Slovenia
by Danijela Černe, Peter Hostnik and Ivan Toplak
Viruses 2021, 13(3), 405; https://doi.org/10.3390/v13030405 - 4 Mar 2021
Cited by 7 | Viewed by 3387
Abstract
Sylvatic rabies was present in Slovenia between 1973 and 2013, with the red fox as the main reservoir of the rabies virus. The first oral rabies vaccination (ORV) control program in foxes started in 1988, using the manual distribution of baits. Significant improvement [...] Read more.
Sylvatic rabies was present in Slovenia between 1973 and 2013, with the red fox as the main reservoir of the rabies virus. The first oral rabies vaccination (ORV) control program in foxes started in 1988, using the manual distribution of baits. Significant improvement of fox vaccination was achieved with the aerial distribution of baits, starting in 1995 and successfully finished with the final, fifty-ninth vaccination campaign in 2019. Between 1979 and 2019, a total of 86,471 samples were tested, and 10,975 (12.69%) rabies-positive animals were identified. Within the ORV, two different vaccines were used, containing modified live virus strain Street Alabama Dufferin (SAD) B19 and SAD Bern, while the last ORV campaigns were completed in 2019, with a vaccine containing a genetically modified strain of SPBN GASGAS. Molecular epidemiological studies of 95 rabies-positive samples, originating from red foxes, badgers, cattle, dogs, martens, cats, and horses, revealed a low genetic diversity of circulating strains and high similarity to strains from neighboring countries. During the elimination program, few vaccine-induced rabies cases were detected: three in red foxes and one case in a marten, with no epidemiological relevance. Slovenia has been officially declared a country free of rabies since 2016. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
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Other

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12 pages, 1453 KiB  
Case Report
Detection of Laryngotracheitis Virus in Poultry Flocks with Respiratory Disorders in Slovenia
by Olga Zorman Rojs, Alenka Dovč, Uroš Krapež, Zoran Žlabravec, Joško Račnik and Brigita Slavec
Viruses 2021, 13(4), 707; https://doi.org/10.3390/v13040707 - 19 Apr 2021
Cited by 9 | Viewed by 3275
Abstract
Infectious laryngotracheitis (ILT) is an acute, highly contagious infectious disease of the upper respiratory tract in chickens and other poultry species that causes significant economic losses in countries worldwide. Between 2017 and 2019, seven outbreaks of mild to severe respiratory disorders with high [...] Read more.
Infectious laryngotracheitis (ILT) is an acute, highly contagious infectious disease of the upper respiratory tract in chickens and other poultry species that causes significant economic losses in countries worldwide. Between 2017 and 2019, seven outbreaks of mild to severe respiratory disorders with high suspicion of ILT occurred in commercial and backyard poultry flocks in Slovenia. In all submissions, infection with ILT virus (ILTV) was confirmed by PCR, which is the first report of ILT in Slovenia. Circulating ILT strains were characterized by the sequence and phylogenetic analysis of two fragments of the ICP4 gene. Four strains—three detected in non-vaccinated flocks and one in a flock vaccinated against ILT—were identical or very similar to the chicken embryo–origin live virus vaccines, and the other three were closely related to Russian, Chinese, Australian, and American field strains and to tissue culture origin vaccine strains. As in other diseases, coinfections with other respiratory pathogens in confirmed ILT cases may cause a more severe condition and prolong the course of the disease. In our study, coinfections with Mycoplasma synoviae (7/7 tested flocks), infectious bronchitis virus (5/5 tested flocks), Mycoplasma gallisepticum (4/7 tested flocks), Ornithobacterium rhinotracheale (3/4 tested flocks), and avian pox virus (1/2 tested flocks) were confirmed, indicating the importance of these pathogens in the occurrence of ILT infections. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Viruses Research in Slovenia)
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