Pigeon circoviruses (PiCV) had been identified worldwide and are responsible for immune suppression and a variety of diseases collectively referred to as young pigeon disease syndrome. Samples from racing pigeons were collected throughout Hungary and analyzed for the presence of PiCV by polymerase chain reaction. The capsid protein coding gene was amplified from ten PiCVs of different origins, and compared with known PiCV sequences. The results indicated that PiCV was highly variable, the viruses formed five distinct genetic groups. Differences of the 3' end of the gene suggested the possibility of genetic recombination among these groups.
Porcine hokovirus (PHoV), a newly discovered member of the family Parvoviridae and the proposed genus Hokovirus, is considered phylogenetically distinct from other parvoviruses. Here, we report a comprehensive spatio-temporal study of PHoV infection in Romanian wild boars. The prevalence of PHoV differed significantly in samples from 2006/2007 (22.76%) and 2010/2011 (50.54%), and also increased with age. Sequence analysis of PHoVs from 2006/2007 showed a close relationship to PHoVs from pigs from England and wild boars from Germany, while the PHoVs from 2010/2011 were mostly similar to isolates from Hong Kong. The most variable regions were detected in the NS1 gene and proved to be suitable for analysis of the genetic diversity of the virus. It was observed that PHoVs from older wild boar samples differed from those collected recently. These results suggested that porcine hokovirus could be a newly emerging virus of both domestic and wild pigs with yet unknown implications.
Novel porcine parvoviruses showing the genetic characteristics of bocaviruses have recently been identified. The first such porcine bocavirus (PoBoV1), described as boca-like virus (PBo-likeV), was discovered in PMWS affected pigs in Sweden. Later, several other bocaviruses with divergent genomes were reported under various names in domestic pigs. This is the first report of the presence of bocaviruses in European wild boars. 842 wild boar samples originating from the Western region of Romania (Transylvania) were collected during the 2006/2007 and the 2010/2011 hunting seasons and tested for the presence of PoBoV1 by polymerase chain reaction and sequencing. The results showed 12.94% (109/842) overall positivity, with an increasing prevalence from the 2006/2007 (9.14%, 43/470) to the 2010/2011 (17.74%, 66/372) season (P < 0.01). Differences between the prevalence of the virus in 6-12-month-old-animal (77.06%, 84/109) and 12-36-month-old-animal (22.94%, 25/109) (P < 0.01) indicated that the infection occurred mainly in younger pigs. Comparative sequence analysis of partial VP1/2 genes from wild boars and those available in the GenBank showed only minor differences, indicating that PoBoV1 circulating within the wild boar populations and domestic pigs from different geographic regions were highly similar.
Potential porcine circovirus type 2 (PCV2) capsid protein epitopes, suitable for expression on the surface of cucumber mosaic virus (CMV) particles were determined by a thorough analysis of the predicted PCV capsid protein structure. The ab initio protein structure prediction was carried out with fold recognition and threading methods. The putative PCV epitopes were selected on the basis of PCV virion models and integrated into the plant virus coat protein, after amino acid position 131. The recombinants were tested for infectivity and stability on different Nicotiana species and stable recombinant virus particles were purified. The particles were tested for their ability to bind to PCV induced porcine antibodies and used for specific antibody induction in mice and pigs. The results showed that PCV epitopes expressed on the CMV surface were recognized by the porcine antibodies and they were also able to induce PCV specific antibody response. Challenge experiment with PCV2 carried out in immunized pigs showed partial protection against the infection. Based on these results it was concluded that specific antiviral vaccine production for the given pathogen was feasible, offering an inexpensive way for the mass production of such vaccines.
PRRs are sentinels of the innate immune system, with TLRs being the most important. Assays for TLR ligand interactions have been used to gain insights into their function and signaling pathways. As significant differences exist between species with regard to ligand recognition, it is necessary to adapt these tools for TLRs of other species. In the present work, we describe a species-specific cell-based assay adapted for the analysis of single PRRs. Human embryonic kidney 293T cells were stably transfected with the NF-κB-inducible reporter gene secreted embryonic alkaline phosphatase (SEAP) together with bovine TLR2. We compared the SEAP response with an existing luciferase NF-κB reporter assay for correlation with IL-8 production. A dose-dependent response was detected upon stimulation using both methods with good correlation to IL-8 secretion. Lower stimulant concentrations were detected by SEAP assay than IL-8 secretion. The luciferase assay produced high non-specific background for all ligand concentrations. Of all assays tested, we found the bovine-specific SEAP reporter assay to be the most convenient and delivered results in the shortest time. The developed reporter cell line would lend well to rapid, high-throughput TLR ligand screening for cattle.
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