[Show abstract][Hide abstract] ABSTRACT: Knowledge of bornaviruses has expanded considerably during the last decade. A possible reservoir of mammalian Borna disease virus has been identified, divergent bornaviruses have been detected in birds and reptiles, and endogenous bornavirus-like elements have been discovered in the genomes of vertebrates of several species. Previous sequence comparisons and alignments have indicated that the members of the current family Bornaviridae are phylogenetically diverse and are not adequately classified in the existing bornavirus taxonomy supported by the International Committee on Taxonomy of Viruses (ICTV). We provide an update of these analyses and describe their implications for taxonomy. We propose retaining the family name Bornaviridae and the genus Bornavirus but reorganizing species classification. PAirwise Sequence Comparison (PASC) of bornavirus genomes and Basic Local Alignment Search Tool (BLAST) comparison of genomic and protein sequences, in combination with other already published phylogenetic analyses and known biological characteristics of bornaviruses, indicate that this genus should include at least five species: Mammalian 1 bornavirus (classical Borna disease virus and divergent Borna disease virus isolate No/98), Psittaciform 1 bornavirus (avian/psittacine bornaviruses 1, 2, 3, 4, 7), Passeriform 1 bornavirus (avian/canary bornaviruses C1, C2, C3, LS), Passeriform 2 bornavirus (estrildid finch bornavirus EF), and Waterbird 1 bornavirus (avian bornavirus 062CG). This classification is also in line with biological characteristics of these viruses and their vertebrate hosts. A snake bornavirus, proposed to be named Loveridge’s garter snake virus 1, should be classified as a member of an additional species (Elapid 1 bornavirus), unassigned to a genus, in the family Bornaviridae. Avian bornaviruses 5, 6, MALL, and another “reptile bornavirus” (“Gaboon viper virus”) should stay unclassified until further information becomes available. Finally, we propose new virus names and abbreviations when necessary to achieve clear differentiation and unique identification.
Archives of Virology 12/2014; · 2.28 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In this study the first complete sequence of the West Nile virus (WNV) lineage 2 strain currently circulating in Romania was determined. The virus was detected in a Hyalomma marginatum marginatum tick collected from a juvenile song thrush (Turdus philomelos) in the Romanian Danube Delta close to the city of Tulcea, end of August 2013. Our finding emphasizes the role of ticks in introduction and maintenance of WNV infections. Sequence analyses revealed close genetic relationship of the Romanian WNV strain to strain Reb_Volgograd_07_H, which was isolated from human brain tissue during an outbreak of West Nile neuroinvasive disease (WNND) in Russia in 2007. In 2010 the Eastern European lineage 2 WNV caused an outbreak of human WNND in Romania. Partial sequences from subsequent years demonstrated that this WNV strain became endemic in Eastern Europe and has been causing outbreaks of varying sizes in southern Russia since 2007 and in Romania since 2010.
PLoS ONE 10/2014; 9(10):e109905. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Borna disease (BD) is a sporadic neurologic disease of horses and sheep caused by mammalian Borna disease virus (BDV). Its unique epidemiological features include: limited occurrence in certain endemic regions of central Europe, yearly varying disease peaks, and a seasonal pattern with higher disease frequencies in spring and a disease nadir in autumn. It is most probably not directly transmitted between horses and sheep. All these features led to the assumption that an indigenous virus reservoir of BDV other than horses and sheep may exist. The search for such a reservoir had been unsuccessful until a few years ago five BDV-infected shrews were found in a BD-endemic area in Switzerland. So far, these data lacked further confirmation. We therefore initiated a study in shrews in endemic areas of Germany. Within five years 107 shrews of five different species were collected. BDV infections were identified in 14 individuals of the species bicolored white-toothed shrew (Crocidura leucodon, HERMANN 1780), all originating from BD-endemic territories. Immunohistological analysis showed widespread distribution of BDV antigen both in the nervous system and in epithelial and mesenchymal tissues without pathological alterations. Large amounts of virus, demonstrated by presence of viral antigen in epithelial cells of the oral cavity and in keratinocytes of the skin, may be a source of infection for natural and spill-over hosts. Genetic analyses reflected a close relationship of the BDV sequences obtained from the shrews with the regional BDV cluster. At one location a high percentage of BDV-positive shrews was identified in four consecutive years, which points towards a self-sustaining infection cycle in bicolored white-toothed shrews. Analyses of behavioral and population features of this shrew species revealed that the bicolored white-toothed shrew may indeed play an important role as an indigenous host of BDV.
PLoS ONE 04/2014; 9(4):e93659. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The primary structures of host-defense peptides present in frog skin secretions constitute useful molecular markers for establishing taxonomic classifications and investigating phylogenetic relationships between species within a particular genus. Peptidomic analysis has led to the characterization of multiple host-defense peptides in norepinephrine-stimulated skin secretions of three species of frogs from the family Ranidae: Lithobates forreri (Boulenger, 1883), Hylarana luctuosa (Peters, 1871), and Hylarana signata (Günther, 1872). The L. forreri secretions contain ranatuerin-2 (2 peptides), brevinin-1 (4 peptides), and temporin (1 peptide). The H. luctuosa secretions contain brevinin-2 (4 peptides), esculentin-1 (1 peptide), esculentin-2 (1 peptide), palustrin-2 (2 peptides), and temporin (2 peptides). The H. signata secretions contain brevinin-2 (4 peptides), brevinin-1 (5 peptides), palustrin-2 (1 peptide), and temporin (2 peptides). Cladistic analysis based upon the primary structures of 44 ranatuerin-2 peptides from 20 Lithobates species indicates a close phylogenetic relationship between L. forreri, Lithobates onca, and Lithobates yavapaiensis. A similar cladistic analysis based upon the primary structures of 27 brevinin-2 peptides from 8 Hylarana species provides support for a close phylogenetic relationship between H. signata and Hylarana picturata, while showing that the species are not conspecific, with H. luctuosa more distantly related.
Comparative Biochemistry and Physiology Part D Genomics and Proteomics 01/2014; 9C:49-57. · 2.82 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To date the influence of herpesviruses on the development of equine ocular diseases has not been clearly determined.
The purpose of this study was to illustrate the course of equine ocular findings over a period of 18 months at 6 month intervals, in correlation with the results of herpesvirus detection.
266 Lipizzaners in 3 federal states of Austria underwent complete ophthalmologic examination 4 times. Blood samples, nasal- and conjunctival swabs were obtained at the same time and used for the detection of the equid gammaherpesviruses EHV-2 and EHV-5 using consensus herpesvirus PCR and type-specific qPCRs. Ophthalmic findings and results of herpesvirus PCRs were recorded and statistically analysed using one-way ANOVA, and multiple logistic regression analysis to determine the influence of herpesvirus infections and other contributing factors on the presence of ophthalmic findings.
In the first, second, third and fourth examination period 266, 261, 249 and 230 horses were included, respectively. Ophthalmic findings consistent with herpesvirus infections included conjunctival- and corneal pathologies. Statistical analysis revealed that the probability of positive herpesvirus PCR results decreased with progressing age; however the presence of corneal findings increased over time. At the time of each examination 45.1%, 41.8%, 43.0%, and 57.0% of horses with conjunctival or corneal findings, respectively, were positive for EHV-2 and/or EHV-5. However, 31.6%, 17.6%, 20.1%, and 13.0% of clinically sound horses were positive for these herpesviruses at each examination period, too.
Based on the results of our study there is a significant influence of young age on EHV-2 and/or EHV-5 infection. Corneal pathologies increased over time and with progressing age. Whether the identified findings were caused by herpesviruses could not be unequivocally determined.
PLoS ONE 11/2013; 8(11):e79888. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to assess the extent of asinine herpesvirus (AsHV) type 5 infection in 'closed' populations of clinically normal Lipizzaner horses. Peripheral blood mononuclear cells plus nasal and conjunctival swabs were obtained on four occasions over an 18month period from 266 animals as part of a health surveillance programme. Sequence analysis of samples that were positive by nested consensus herpesvirus PCR but negative using quantified equid herpesvirus (EHV) type 2 and 5 PCR, revealed a total of 51 samples from 39 horses positive for AsHV-5. No statistically significant association between animal age, gender or geographical location and infection status was identified. The findings suggest sub-clinical AsHV-5 infection may be encountered more frequently than previously reported.
The Veterinary Journal 11/2013; · 2.17 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Barkedji virus, named after the area of its first identification in Senegal, is a newly discovered flavivirus, for which we propose the abbreviation BJV. In the present study, we report the first-time detection of BJV in Culex perexiguus mosquitoes in Israel in 2011 and determination of its almost complete polyprotein gene sequence. We characterized the BJV genome and defined putative mature proteins, conserved structural elements, and potential enzyme motifs along the polyprotein precursor. By comparing polyproteins and individual proteins of BJV with several other flaviviruses, a distant relationship of BJV to Nounane virus, a recently described African flavivirus, is demonstrated. Phylogenetic analysis of 55 selected flaviviral polyprotein gene sequences exhibits two major clusters, one made up of the classical three clades of flaviviruses: mosquito-borne, tick-borne, and such without known vectors. The other cluster exclusively contains so-called 'insect-specific' flaviviruses, which do not replicate in vertebrate cells. Based on our phylogenetic analysis, BJV is related to other members of the mosquito-borne clade with yet unknown vertebrate hosts, such as Nounane virus, Donggang virus, Chaoyang virus, and Lammi virus. However, with a maximum identity of only 54% to Nounane virus BJV represents a distinct new virus species.
Journal of General Virology 08/2013; · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Sedlec virus (SEDV) was isolated from the blood of a reed warbler (Acrocephalus scirpaceus) in July 1984 in South Moravia, Czech Republic. In this study first genetic data of SEDV are presented which allow an estimate on its phylogenetic and taxonomic positioning within the genus Orthobunyavirus. The phylogenetic analysis of a 369 nt long stretch within the S segment (nucleocapsid protein gene and non-structural S protein gene) indicates genetic relatedness of SEDV to Leanyer virus and Simbu group viruses, while the phylogenetic tree based on 1796 nt long sequences of the L segment (RNA-dependent RNA polymerase gene) demonstrates genetic relationship of SEDV to two yet unclassified orthobunyaviruses: I612045 virus (isolated in India in 1961) and Oyo virus (isolated in Nigeria in 1964). Considering the genetic distances and the phylogenetic analyses, SEDV might represent a novel serogroup of the Orthobunyavirus genus.
Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 08/2013; · 3.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The prevalence of linear keratopathy with progressing age in a closed population of a single horse breed is reported. All Lipizzaners in three federal states in Austria underwent complete ophthalmic examination four times over a period of 18 months, with six-month intervals. Findings consistent with linear keratopathy were recorded, and associated with factors such as sex, location, boarding system and level of performance throughout the study period. Logistic regression was applied to determine the influence of age on ophthalmic findings. On the first, second, third and fourth examinations, 0.8 per cent, 3.1 per cent, 4.4 per cent and 4.8 per cent (of 266, 261, 249 and 230 horses, respectively) of the study population, were diagnosed with linear keratopathy. This finding was consistently identified in the same horses, and once identified, no further progression was noted. Horses with this finding had no history of previous ocular problems or concurrent ocular abnormalities. Statistical analysis did not reveal any influence of sex, location, boarding, or level of performance; however the prevalence of linear keratopathy was found to increase with progressive age (P<0.5). The results of this study indicate that linear keratopathy was not congenital and was non-progressive in the Lipizzaner over a period of 18 months.
[Show abstract][Hide abstract] ABSTRACT: On July 15, 2012, adult Anopheles hyrcanus (Pallas 1771) mosquitoes were caught next to a farm barn near Rust, Burgenland, close to Lake Neusiedl National Park in eastern Austria. Six weeks later, adults of this invasive species were also found in a sheep shelter outside the village of Oggau and another 2 weeks later, in a horse barn in Mörbisch. The morphological typing was confirmed genetically by amplification and sequencing of a 1,404-bp-long fragment within the 5.8S ribosomal RNA gene, the internal transcribed spacer 2, and the 28S ribosomal RNA gene. Out of two A. hyrcanus pools analyzed, one was found positive for Plasmodium sp. A 460-bp-long sequence within the mitochondrial cytochrome b region revealed 100 % identity to a sequence of a Plasmodium parasite identified in a New Zealand bellbird (Anthornis melanura). The Austrian finding sites are close to the Hungarian border. In Hungary, the occurrence of A. hyrcanus was already reported in 1963. A. hyrcanus is considered the most important potential vector of malaria in southern France today. In Austria, sporadic autochthonous malaria cases could emerge, caused by immigration from malaria-endemic countries and heavy tourism. However, the broad population coverage of the Austrian health care system makes the reestablishment of endemic areas for malaria unlikely.
Wiener klinische Wochenschrift 02/2013; · 0.79 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A herd of Lipizzaners (n=266) in three locations [Federal stud Piber (Styria), Heldenberg (Lower Austria) and the Spanish Riding School Vienna (Vienna)] were examined for ocular diseases and tested for herpesviruses (HVs) in PBMCs, nasal- and conjunctival swabs. In 167 (62.8%) horses equid herpesvirus-2 (EHV-2) nucleic acid was identified in at least one of the investigated samples, and 136 (51.1%) horses were positive for equid herpesvirus-5 (EHV-5) nucleic acid in at least one of the samples. One hundred and five (39.5%) of the horses mentioned above were identified with EHV-2/EHV-5 double infections. The results were correlated with potential influence factors such as age, gender, location, and housing. Statistical analysis demonstrated a significantly higher percentage of horses diagnosed with HV infections stabled in one location (Piber); also, housing in free stalls was associated with a higher rate of HV infections. Horses carrying EHV-2 or EHV-5 nucleic acid were significantly younger than the negative group. Statistical analysis of PCR results and ophthalmic findings did not prove a significant association between any disease pattern and presence of the equid gammaherpesviruses EHV-2 and EHV-5. The results of this herd survey indicate that horses younger than eight years of age, housed in free stalls with high population densities have a significantly increased risk for infection with EHV-2 and EHV-5. However no prediction can be made whether this results in ophthalmic disease. This is the first ophthalmic-virologic survey of this magnitude in a single horse breed.
[Show abstract][Hide abstract] ABSTRACT: Proventricular dilatation disease (PDD) is a fatal, progressive neurological disorder of psittacine birds, which is caused by a single-stranded RNA virus, the avian bornavirus (ABV). The disease pattern includes lymphoplasmacytic inflammation of the central, peripheral and autonomic nervous system. Seven avian bornavirus genotypes have been identified during the last years. So far only monoinfections with a single genotype of ABV have been attributed to PDD cases. However, after a recent survey discovered a case of a double infection with two different ABV genotypes, this seemed to indicate the need for a more systematic search for mixed infections. Brain specimens from 21 psittacine birds affected with PDD were examined. Aim of the investigation was to generate partial ABV sequences of a part of the matrix protein (M) gene and to evaluate whether sequences of more than one ABV genotype were present. RNA was extracted, and subjected to reverse transcriptase PCR with primer pairs generating a partial sequence of the matrix protein (M) gene, followed by a cloning procedure. Ten clones per case were sequenced in order to elucidate whether sequences characteristic for one or more than one genotype were present. In 19 of 21 cases clear M gene sequences could be generated; in two cases nucleic acid amplification failed. Seven birds were infected with ABV 2 and nine with ABV 4, representing the predominant genotypes in Europe. Two cases showed a mixed infection with ABV 2 and ABV 4, and one case a mixed infection with ABV 2 and ABV 6. These results suggest that the molecular cloning method is a useful tool for distinguishing between single and multiple infection events by different ABV genotypes.
[Show abstract][Hide abstract] ABSTRACT: Aujeszky's disease (AD), caused by Suid herpesvirus type 1 (SuHV-1), is an economically important disease in domestic swine. Thus, rigorous control programmes have been implemented and consecutively AD in domestic swine was successfully eradicated in many countries, including Austria. However, SuHV-1 continues to thrive in wild boar populations, as indicated by high seroprevalences in a number of European countries and by occasional cases of AD in hunting dogs. For the first time, SuHV-1 was detected in Austrian wild boar and a molecular characterization of SuHV-1 isolated from wild boar and hunting dogs was performed. Results of preliminary serological analyses suggest a regional SuHV-1 seroprevalence of over 30% in free-living and almost 70% in fenced wild boar from Eastern Austria. Molecular typing of Austrian SuHV-1 isolates of wild boar origin revealed the presence of two genetically distinct variants of SuHV-1, both capable of infecting dogs that have been exposed to infected wild boar during hunting.
[Show abstract][Hide abstract] ABSTRACT: Cowpox virus (CPXV) is described as the source of the first vaccine used to prevent the onset and spread of an infectious disease. It is one of the earliest described members of the genus Orthopoxvirus, which includes the viruses that cause smallpox and monkeypox in humans. Both the historic and current literature describe "cowpox" as a disease with a single etiologic agent. Genotypic data presented herein indicate that CPXV is not a single species, but a composite of several (up to 5) species that can infect cows, humans, and other animals. The practice of naming agents after the host in which the resultant disease manifests obfuscates the true taxonomic relationships of "cowpox" isolates. These data support the elevation of as many as four new species within the traditional "cowpox" group and suggest that both wild and modern vaccine strains of Vaccinia virus are most closely related to CPXV of continental Europe rather than the United Kingdom, the homeland of the vaccine.
PLoS ONE 08/2011; 6(8):e23086. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Populations of the Chiricahua leopard frog Lithobates chiricahuensis (Ranidae) occupying regions in southern Arizona (southern range) are morphologically distinct from those from the Mogollon Rim of central Arizona (northern range) and a comparison of DNA sequences of mitochondrial genes has suggested that they may represent separate species. Peptidomic analysis of norepinephrine-stimulated skin secretions has led to the identification of six peptides with antimicrobial activity in samples from specimens from both groups. The primary structure of the peptides (esculentin-2 CHa, ranatuerin-2 CHa, -CHb, and -CHc, and brevinin-1 CHa and -CHb) isolated from both southern and northern range frogs are identical consistent with the proposal that the two populations are conspecific. However, palustrin-2CHa and the atypical brevinin-1 CHc (FFPTIAG*****LTKLFCA ITKKC), containing a five amino acid residue deletion, were identified only in secretions from southern range specimens. Consequently, there is some support for the proposal that the two populations are closely related but separate species but this support is relatively weak. Esculentin-2 CHa (GFSSIFRGVAKFASKGLG KDLAKLGVDLVACKISKQC) displayed the highest antimicrobial potency (MIC ≤ 10μM) against a variety of microorganisms and was only moderately hemolytic (LC(50) = 150 μM). Cladistic analysis based upon the primary structures of brevinin-1 peptides indicates a close phylogenetic relationship between L. chiricahuensis, L. onca, and L. yavapaiensis.