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Granberg F.,Swedish University of Agricultural Sciences | Karlsson O.E.,Swedish University of Agricultural Sciences | Belak S.,Swedish University of Agricultural Sciences | Belak S.,The National Veterinary Institute SVA
Methods in Molecular Biology | Year: 2014

Metagenomic approaches have become invaluable for culture-independent and sequence-independent detection and characterization of disease-associated pathogens. Here, the sequential steps from sampling to verification of results are described for a metagenomic-based approach to detect potential pathogens in honeybees. The pre-sequencing steps are given in detail, but due to the rapid development of sequencing technologies, all platform-specific procedures, as well as subsequent bioinformatics analysis, are more generally described. It should also be noted that this approach could, with minor modifications, be adapted for other organisms and sample matrices. © Springer Science+Business Media New York 2015. Source

Cholleti H.,The National Veterinary Institute SVA | Cholleti H.,Uppsala University | Paidikondala M.,The National Veterinary Institute SVA | Paidikondala M.,University of Helsinki | And 3 more authors.
Virus Research | Year: 2013

Equine arteritis virus (EAV) causes a respiratory and reproductive disease in horses, equine viral arteritis. Though cell death in infection with EAV is considered to occur by apoptosis, the underlying molecular mechanism has not been extensively elucidated. We investigated the expression of mRNA of pro-apoptotic and caspase genes during EAV infection in BHK21 cells, a well-established cell type for EAV replication. Using a SYBR Green real-time PCR, mRNA of p53, Bax, caspase 3 and caspase 9 were found up-regulated in a time dependent manner in EAV infected cells. Western blot analysis for caspase 3 and caspase 9 showed expression of cleaved forms of these proteins during EAV infection. In addition, a luminescence-based cell assay for caspase 3/7 activation as a hallmark in apoptosis confirmed apoptotic cell death. The findings demonstrate that cell death in EAV infected BHK21 cells results from apoptosis mediated through the intrinsic signalling pathway. © 2012 Elsevier B.V. Source

Metreveli G.,Swedish University of Agricultural Sciences | Wagberg L.,Uppsala University | Emmoth E.,The National Veterinary Institute SVA | Belak S.,The National Veterinary Institute SVA | And 3 more authors.
Advanced Healthcare Materials | Year: 2014

This is the first time a 100% natural, unmodified nanofibrous polymer-based membrane is demonstrated capable of removing viruses solely based on the size-exclusion principle, with a log10 reduction value (LRV) ≥ 6.3 as limited by the assay lower detection limit and the feed virus titre, thereby matching the performance of industrial synthetic polymer virus removal filters. © 2014 The Authors. Source

Widen F.,The National Veterinary Institute SVA | Everett H.,Animal Health and Veterinary Laboratories Agency AHVLA | Blome S.,Institute of Diagnostic Virology | Fernandez Pinero J.,Research Center en Sanidad Animal | And 5 more authors.
Research in Veterinary Science | Year: 2014

Classical swine fever is one of the most important infectious diseases for the pig industry worldwide due to its economic impact. Vaccination is an effective means to control disease, however within the EU its regular use is banned owing to the inability to differentiate infected and vaccinated animals, the so called DIVA principle. This inability complicates monitoring of disease and stops international trade thereby limiting use of the vaccine in many regions. The C-strain vaccine is safe to use and gives good protection. It is licensed for emergency vaccination in the EU in event of an outbreak. Two genetic assays that can distinguish between wild type virus and C-strain vaccines have recently been developed. Here the results from a comparison of these two real-time RT-PCR assays in an interlaboratory exercise are presented. Both assays showed similar performance. © 2014 Elsevier Ltd. All rights reserved. Source

Van Borm S.,Veterinary and Agrochemical Research Center | Belak S.,Swedish University of Agricultural Sciences | Belak S.,The National Veterinary Institute SVA | Freimanis G.,The Pirbright Institute | And 7 more authors.
Methods in Molecular Biology | Year: 2014

The development of high-throughput molecular technologies and associated bioinformatics has dramatically changed the capacities of scientists to produce, handle, and analyze large amounts of genomic, transcriptomic, and proteomic data. A clear example of this step-change is represented by the amount of DNA sequence data that can be now produced using next-generation sequencing (NGS) platforms. Similarly, recent improvements in protein and peptide separation efficiencies and highly accurate mass spectrometry have promoted the identification and quantification of proteins in a given sample. These advancements in biotechnology have increasingly been applied to the study of animal infectious diseases and are beginning to revolutionize the way that biological and evolutionary processes can be studied at the molecular level. Studies have demonstrated the value of NGS technologies for molecular characterization, ranging from metagenomic characterization of unknown pathogens or microbial communities to molecular epidemiology and evolution of viral quasispecies. Moreover, high-throughput technologies now allow detailed studies of hostpathogen interactions at the level of their genomes (genomics), transcriptomes (transcriptomics), or proteomes (proteomics). Ultimately, the interaction between pathogen and host biological networks can be questioned by analytically integrating these levels (integrative OMICS and systems biology). The application of high-throughput biotechnology platforms in these fields and their typical low-cost per information content has revolutionized the resolution with which these processes can now be studied.The aim of this chapter is to provide a current and prospective view on the opportunities and challenges associated with the application of massive parallel sequencing technologies to veterinary medicine, with particular focus on applications that have a potential impact on disease control and management. © Springer Science+Business Media New York 2015. Source

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