| 1. |
- Belak, Sandor, et al.
(författare)
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New viruses in veterinary medicine, detected by metagenomic approaches
- 2013
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Ingår i: Veterinary Microbiology. - : Elsevier BV. - 0378-1135 .- 1873-2542. ; 165, s. 95-101
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Tidskriftsartikel (refereegranskat)abstract
- In our world, which is faced today with exceptional environmental changes and dramatically intensifying globalisation, we are encountering challenges due to many new factors, including the emergence or re-emergence of novel, so far “unknown” infectious diseases. Although a broad arsenal of diagnostic methods is at our disposal, the majority of the conventional diagnostic tests is highly virus-specific or is targeted entirely towards a limited group of infectious agents. This specificity complicates or even hinders the detection of new or unexpected pathogens, such as new, emerging or re-emerging viruses or novel viral variants. The recently developed approaches of viral metagenomics provide an effective novel way to screen samples and detect viruses without previous knowledge of the infectious agent, thereby enabling a better diagnosis and disease control, in line with the “One World, One Health” principles (www.oneworldonehealth.org). Using metagenomic approaches, we have recently identified a broad variety of new viruses, such as novel bocaviruses, Torque Teno viruses, astroviruses, rotaviruses and kobuviruses in porcine disease syndromes, new virus variants in honeybee populations, as well as a range of other infectious agents in further host species. These findings indicate that the metagenomic detection of viral pathogens is becoming now a powerful, cultivation-independent, and useful novel diagnostic tool in veterinary diagnostic virology.
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| 2. |
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| 3. |
- Berg, Mikael, et al.
(författare)
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Single primer isothermal amplification (SPIA) combined with next generation sequencing provides complete bovine coronavirus genome coverage and higher sequence depth compared to sequence-independent single primer amplification (SISPA)
- 2017
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 12
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Tidskriftsartikel (refereegranskat)abstract
- Coronaviruses are of major importance for both animal and human health. With the emergence of novel coronaviruses such as SARS and MERS, the need for fast genome characterisation is ever so important. Further, in order to understand the influence of quasispecies of these viruses in relation to biology, techniques for deep-sequence and full-length viral genome analysis are needed. In the present study, we compared the efficiency of two sequence-independent approaches [sequence-independent single primer amplification (SISPA) and single primer isothermal amplification (SPIA, represented by the Ovation kit)] coupled with high throughput sequencing to generate the full-length genome of bovine coronavirus (BCoV) from a nasal swab. Both methods achieved high genome coverage (100% for SPIA and 99% for SISPA), however, there was a clear difference in the percentage of reads that mapped to BCoV. While approximately 45% of the Ovation reads mapped to BCoV (sequence depth of 169-284 944), only 0.07% of the SISPA reads (sequence depth of 0-249) mapped to the reference genome. Although BCoV was the focus of the study we also identified a bovine rhinitis B virus (BRBV) in the data sets. The trend for this virus was similar to that observed for BCoV regarding Ovation vs. SISPA, but with fewer sequences mapping to BRBV due to a lower amount of this virus. In summary, the SPIA approach used in this study produced coverage of the entire BCoV (high copy number) and BRBV (low copy number) and a high sequence/genome depth compared to SISPA. Although this is a limited study, the results indicate that the Ovation method could be a preferred approach for full genome sequencing if a low copy number of viral RNA is expected and if high sequence depth is desired.
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| 4. |
- Björnström, Agnes, et al.
(författare)
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Sindbis virus neutralising antibodies detected in Swedish horses
- 2021
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Ingår i: One Health. - : Elsevier. - 2352-7714. ; 12
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Tidskriftsartikel (refereegranskat)abstract
- A number of viruses transmitted by mosquitoes are well known to cause disease in both humans and horses, ranging from mild fevers to mortal neurological disease. A recently discovered connection between the alphavirus Sindbis virus (SINV) and neurological disease in horses in South Africa initiated this serological study in northern Europe, where the same genotype of SINV (SINV-I) is also highly endemic. We tested 171 serum samples, originally obtained from horses for other reasons from April to October 2019, for presence of SINV neutralising antibodies using a plaque reduction neutralisation test (PRNT). The serum from six horses reduced the plaque count more than 80%, and two out of these reduced the plaque count more than 90%. These horses were sampled in six different regions of Sweden, and included individuals sampled from April to August. This study shows that horses in Sweden have become infected with SINV and developed neutralising antibodies. Potential connections between infection and development of disease are important questions for future studies.
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| 5. |
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| 6. |
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| 7. |
- Blomström, Anne-Lie
(författare)
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Applications of viral metagenomics in the veterinary field : looking for the unknown
- 2010
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Viral metagenomics provide a powerful technology to investigate the viral flora of healthy and sick animals. Using these methodologies, we gain a better understanding in the etiology of diseases, as well as deepen our knowledge into the viruses circulating in nature and the complex interaction between virus and host. The aim of this thesis was to utilize viral metagenomics into different areas of interest for veterinary science. One multifactorial disease complex was studied, as were a disease of unknown etiology, where traditional methods had failed to identify a causative agent. The final study was on viruses in an arthropod vector. Postweaning multisystemic wasting syndrome (PMWS) was the multifactorial disease chosen here. Though, porcine circovirus type 2 (PCV-2) has been found the causative agent other factors, such as viruses, are believed to influence the disease. Using multiple displacement amplification followed by large-scale sequencing we discovered, apart from PCV-2 and torque teno virus, a novel porcine bocavirus in the background of PCV-2 in lymph nodes collected from pigs suffering from PMWS. This co-infection was seen in a high percentage (71%) of pigs suffering from PMWS compared to only 33% in pigs without PMWS. Shaking mink syndrome (SMS) was used to investigate a disease of unknown etiology. By random amplification and large-scale sequencing we found an astrovirus in the brain of minks experimentally infected with brain homogenate from diseased animals. Astrovirus was also detected in the brain of naturally infected minks. Vectors are important transmitters of disease, and in the final study, soft ticks (Ornithodoros) collected from a warthog burrow in Uganda were investigated for the presence of viruses. Among others, we discovered a possible novel RNA virus that showed a distant relationship to hepatitis E virus. In conclusion, viral metagenomics have successfully been applied to investigate three important areas for veterinary science and through these studies three novel viruses were discovered and genetically characterized.
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| 8. |
- Blomström, Anne-Lie, et al.
(författare)
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Astrovirus as a possible cause of congenital tremor type AII in piglets?
- 2014
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Ingår i: Acta Veterinaria Scandinavica. - : Springer Science and Business Media LLC. - 0044-605X .- 1751-0147. ; 56
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Tidskriftsartikel (refereegranskat)abstract
- Background: Congenital tremor is associated with demyelination of the brain and spinal cord and is clinically noted as outbreaks of trembling and shaking in newborn piglets during a limited time-period. Six forms of the disease have been described, where form AII may be caused by an, as yet, unidentified viral infection. This study aimed to investigate the presence of astrovirus and circovirus by sequencing and polymerase chain reaction (PCR) analysis and by relating the findings to the occurrence of disease and lesions in the brain, in 4-6 days-old piglets obtained from a clinical outbreak of congenital tremor.Results: In piglets with congenital tremor, there were mild to moderate vacuolar changes of the white matter in the cerebrum, brain stem and cerebellum. In healthy piglets, less conspicuous vacuolar changes were detected. One healthy and one diseased piglet were positive for porcine circovirus type 2. The nested pan-PCR showed the presence of astrovirus in at least one brain region in all piglets and by sequencing, two different porcine astrovirus lineages were identified.Conclusions: The results do not support previous studies identifying porcine circovirus type 2 as the cause of congenital tremor. The demonstration of astrovirus in the brain of piglets suffering from congenital tremor is interesting. However, astrovirus was demonstrated in both healthy and diseased individuals and therefore, further studies are warranted to determine the possible involvement of astrovirus in the pathogenesis of congenital tremor in pigs.
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| 9. |
- Blomström, Anne-Lie, et al.
(författare)
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Characterisation of the Virome of Tonsils from Conventional Pigs and from Specific Pathogen-Free Pigs
- 2018
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Ingår i: Viruses. - : MDPI AG. - 1999-4915. ; 10
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Tidskriftsartikel (refereegranskat)abstract
- Porcine respiratory disease is a multifactorial disease that can be influenced by a number of different microorganisms, as well as by non-infectious factors such as the management and environment of the animals. It is generally believed that the interaction between different infectious agents plays an important role in regard to respiratory diseases. Therefore, we used high-throughput sequencing combined with viral metagenomics to characterise the viral community of tonsil samples from pigs coming from a conventional herd with lesions in the respiratory tract at slaughter. In parallel, samples from specific pathogen-free pigs were also analysed. This study showed a variable co-infection rate in the different pigs. The differences were not seen at the group level but in individual pigs. Some viruses such as adenoviruses and certain picornaviruses could be found in most pigs, while others such as different parvoviruses and anelloviruses were only identified in a few pigs. In addition, the complete coding region of porcine parvovirus 7 was obtained, as were the complete genomes of two teschoviruses. The results from this study will aid in elucidating which viruses are circulating in both healthy pigs and in pigs associated with respiratory illness. This knowledge is needed for future investigations into the role of viral-viral interactions in relation to disease development.
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| 10. |
- Blomström, Anne-Lie, et al.
(författare)
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Detection of a Novel Astrovirus in Brain Tissue of Mink Suffering from Shaking Mink Syndrome by Use of Viral Metagenomics
- 2010
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Ingår i: Journal of Clinical Microbiology. - 0095-1137 .- 1098-660X. ; 48, s. 4392-4396
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Tidskriftsartikel (refereegranskat)abstract
- In 2000, farmed mink kits in Denmark were affected by a neurological disorder. The characteristic clinical signs included shaking, staggering gait, and ataxia. The disease, given the name shaking mink syndrome, was reproduced by the inoculation of brain homogenate from affected mink kits into healthy ones. However, the etiology remained unknown despite intensive efforts. In this study, random amplification and large-scale sequencing were used, and an astrovirus was detected in the brain tissue of three experimentally infected mink kits. This virus also was found in the brain of three mink kits naturally displaying the disease but not in the six healthy animals investigated. The complete coding region of the detected astrovirus was sequenced and compared to those of both a mink astrovirus associated with preweaning diarrhea and to a recently discovered human astrovirus associated with a case of encephalitis in a boy with x-linked agammaglobulinemia. The identities were 80.4 and 52.3%, respectively, showing that the virus described in this study was more similar to the preweaning diarrhea mink astrovirus. For the nonstructural coding regions the sequence identity was around 90% compared to that of the astrovirus, which is associated with preweaning diarrhea in mink. The region coding for the structural protein was more diverse, showing only 67% sequence identity. This finding is of interest not only because the detected virus may be the etiological agent of the shaking mink syndrome but also because this is one of the first descriptions of an astrovirus found in the central nervous system of animals.
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