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Open AccessArticle

Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy

1
Istituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, 64100 Teramo, Italy
2
NeuroCenter, Department of Clinical Research and Veterinary Public Health, University of Bern, 3012 Bern, Switzerland
3
Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland
4
Unità Operativa Complessa Servizio di Sanità Animale, ASL Pescara, 65100 Pescara, Italy
5
Dipartimento di Medicina Veterinaria, Università degli Studi di Bari, 70010 Valenzano, Italy
*
Author to whom correspondence should be addressed.
Viruses 2020, 12(5), 530; https://doi.org/10.3390/v12050530
Received: 23 April 2020 / Revised: 7 May 2020 / Accepted: 9 May 2020 / Published: 11 May 2020
(This article belongs to the Special Issue Gastroenteritis Viruses)
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.
Keywords: bovine astrovirus; nanopore technology; brain; neurological signs; cattle; phylogeny; ISH; Italy bovine astrovirus; nanopore technology; brain; neurological signs; cattle; phylogeny; ISH; Italy
MDPI and ACS Style

Zaccaria, G.; Lorusso, A.; Hierweger, M.M.; Malatesta, D.; Defourny, S.V.; Ruggeri, F.; Cammà, C.; Ricci, P.; Domenico, M.D.; Rinaldi, A.; Decaro, N.; D’Alterio, N.; Petrini, A.; Seuberlich, T.; Marcacci, M. Detection of Astrovirus in a Cow with Neurological Signs by Nanopore Technology, Italy. Viruses 2020, 12, 530.

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