Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy
In this study, starting from nucleic acids purified from the brain tissue, Nanopore technology was used to identify the etiological agent of severe neurological signs observed in a cow which was immediately slaughtered. Histological examination revealed acute non-suppurative encephalomyelitis affect...
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doaj-cb06c1ee1caa48228c5b7fd6b1e511582020-11-25T02:10:14ZengMDPI AGViruses1999-49152020-05-011253053010.3390/v12050530Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, ItalyGuendalina Zaccaria0Alessio Lorusso1Melanie M. Hierweger2Daniela Malatesta3Sabrina VP Defourny4Franco Ruggeri5Cesare Cammà6Pasquale Ricci7Marco Di Domenico8Antonio Rinaldi9Nicola Decaro10Nicola D’Alterio11Antonio Petrini12Torsten Seuberlich13Maurilia Marcacci14Istituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyNeuroCenter, Department of Clinical Research and Veterinary Public Health, University of Bern, 3012 Bern, SwitzerlandIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyUnità Operativa Complessa Servizio di Sanità Animale, ASL Pescara, 65100 Pescara, ItalyIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyUnità Operativa Complessa Servizio di Sanità Animale, ASL Pescara, 65100 Pescara, ItalyIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyDipartimento di Medicina Veterinaria, Università degli Studi di Bari, 70010 Valenzano, ItalyIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyGraduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, SwitzerlandIstituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, ItalyIn this study, starting from nucleic acids purified from the brain tissue, Nanopore technology was used to identify the etiological agent of severe neurological signs observed in a cow which was immediately slaughtered. Histological examination revealed acute non-suppurative encephalomyelitis affecting the brainstem, cerebrum, cerebellum, and medulla oblongata, while by using PCR-based assays, the nucleic acids of major agents for neurological signs were not detected. By using Nanopore technology, 151 sequence reads were assigned to Bovine Astrovirus (BoAstV). Real-time RT-PCR and in situ hybridization (ISH) confirmed the presence of viral RNA in the brain. Moreover, using the combination of fluorescent ISH and immunofluorescence (IF) techniques, it was possible to detect BoAstV RNA and antigens in the same cells, suggesting the active replication of the virus in infected neurons. The nearly whole genome of the occurring strain (BoAstV PE3373/2019/Italy), obtained by Illumina NextSeq 500, showed the highest nucleotide sequence identity (94.11%) with BoAstV CH13/NeuroS1 26,730 strain, an encephalitis-associated bovine astrovirus. Here, we provide further evidence of the role of AstV as a neurotropic agent. Considering that in a high proportion of non-suppurative encephalitis cases, which are mostly indicative of a viral infection, the etiologic agent remains unknown, our result underscores the value and versatility of Nanopore technology for a rapid diagnosis when the PCR-based algorithm gives negative results.https://www.mdpi.com/1999-4915/12/5/530bovine astrovirusnanopore technologybrainneurological signscattlephylogeny |
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language |
English |
format |
Article |
sources |
DOAJ |
author |
Guendalina Zaccaria Alessio Lorusso Melanie M. Hierweger Daniela Malatesta Sabrina VP Defourny Franco Ruggeri Cesare Cammà Pasquale Ricci Marco Di Domenico Antonio Rinaldi Nicola Decaro Nicola D’Alterio Antonio Petrini Torsten Seuberlich Maurilia Marcacci |
spellingShingle |
Guendalina Zaccaria Alessio Lorusso Melanie M. Hierweger Daniela Malatesta Sabrina VP Defourny Franco Ruggeri Cesare Cammà Pasquale Ricci Marco Di Domenico Antonio Rinaldi Nicola Decaro Nicola D’Alterio Antonio Petrini Torsten Seuberlich Maurilia Marcacci Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy Viruses bovine astrovirus nanopore technology brain neurological signs cattle phylogeny |
author_facet |
Guendalina Zaccaria Alessio Lorusso Melanie M. Hierweger Daniela Malatesta Sabrina VP Defourny Franco Ruggeri Cesare Cammà Pasquale Ricci Marco Di Domenico Antonio Rinaldi Nicola Decaro Nicola D’Alterio Antonio Petrini Torsten Seuberlich Maurilia Marcacci |
author_sort |
Guendalina Zaccaria |
title |
Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy |
title_short |
Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy |
title_full |
Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy |
title_fullStr |
Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy |
title_full_unstemmed |
Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy |
title_sort |
detection of astrovirus in a cow with neurological signs by nanopore technology, italy |
publisher |
MDPI AG |
series |
Viruses |
issn |
1999-4915 |
publishDate |
2020-05-01 |
description |
In this study, starting from nucleic acids purified from the brain tissue, Nanopore technology was used to identify the etiological agent of severe neurological signs observed in a cow which was immediately slaughtered. Histological examination revealed acute non-suppurative encephalomyelitis affecting the brainstem, cerebrum, cerebellum, and medulla oblongata, while by using PCR-based assays, the nucleic acids of major agents for neurological signs were not detected. By using Nanopore technology, 151 sequence reads were assigned to Bovine Astrovirus (BoAstV). Real-time RT-PCR and in situ hybridization (ISH) confirmed the presence of viral RNA in the brain. Moreover, using the combination of fluorescent ISH and immunofluorescence (IF) techniques, it was possible to detect BoAstV RNA and antigens in the same cells, suggesting the active replication of the virus in infected neurons. The nearly whole genome of the occurring strain (BoAstV PE3373/2019/Italy), obtained by Illumina NextSeq 500, showed the highest nucleotide sequence identity (94.11%) with BoAstV CH13/NeuroS1 26,730 strain, an encephalitis-associated bovine astrovirus. Here, we provide further evidence of the role of AstV as a neurotropic agent. Considering that in a high proportion of non-suppurative encephalitis cases, which are mostly indicative of a viral infection, the etiologic agent remains unknown, our result underscores the value and versatility of Nanopore technology for a rapid diagnosis when the PCR-based algorithm gives negative results. |
topic |
bovine astrovirus nanopore technology brain neurological signs cattle phylogeny |
url |
https://www.mdpi.com/1999-4915/12/5/530 |
work_keys_str_mv |
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