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      Changes on the viral capsid surface during the evolution of porcine circovirus type 2 (PCV2) from 2009 till 2018 may lead to a better receptor binding

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          Abstract

          Porcine circovirus type 2 (PCV2) is the primary causative agent of porcine circovirus-associated diseases (PCVAD). Three major PCV2 genotypes (PCV2a, PCV2b, and PCV2d) have been identified globally. Despite their worldwide distribution, the prevalence and genetic evolution of PCV2 in Belgium has not previously been determined. In this study, 319 samples from animals suffering from diseases likely to be associated with PCV2 were collected from 2009 to 2018 and analysed by virus titration. The overall prevalence of PCV2 in PCVAD-suspected cases was 15.7 per cent (50/319). The phylogenetic analysis demonstrated that at least three genotypes (PCV2a, PCV2b, and PCV2d) circulated in Belgium from 2009 till 2018, and that PCV2 evolved from PCV2a to PCV2b and from PCV2d-1 to PCV2d-2. Sequence comparison among the forty-three PCV2 isolates showed that they had 89.7–100 per cent nucleotide-sequence and 88.5–100 per cent amino-acid-sequence identities. Three amino acid sites were under positive selection. Three-dimensional analysis of genotype-specific amino acids revealed that most of the mutations were on the outside of the cap protein with a few conserved mutations present on the inner side. Mutations toward more basic amino acids were found on the upper and tail parts of two connecting capsid proteins which form one big contact region, most probably involved in receptor binding. The lower part was relatively conserved. This polarity change together with the formation of an extruding part drive the virus to a more efficient GAG receptor binding. Taken together, these results showed a genotype shift from PCV2a to PCV2b and later on from PCV2d-1 to PCV2d-2, and a PCV2 evolution toward a better receptor binding capacity.

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          Multiple diverse circoviruses infect farm animals and are commonly found in human and chimpanzee feces.

          Circoviruses are known to infect birds and pigs and can cause a wide range of severe symptoms with significant economic impact. Using viral metagenomics, we identified circovirus-like DNA sequences and characterized 15 circular viral DNA genomes in stool samples from humans in Pakistan, Nigeria, Tunisia, and the United States and from wild chimpanzees. Distinct genomic features and phylogenetic analysis indicate that some viral genomes were part of a previously unrecognized genus in the Circoviridae family we tentatively named "Cyclovirus" whose genetic diversity is comparable to that of all the known species in the Circovirus genus. Circoviridae detection in the stools of U.S. adults was limited to porcine circoviruses which were also found in most U.S. pork products. To determine whether the divergent cycloviruses found in non-U.S. human stools were of dietary origin, we genetically compared them to the cycloviruses in muscle tissue samples of commonly eaten farm animals in Pakistan and Nigeria. Limited genetic overlap between cycloviruses in human stool samples and local cow, goat, sheep, camel, and chicken meat samples indicated that the majority of the 25 Cyclovirus species identified might be human viruses. We show that the genetic diversity of small circular DNA viral genomes in various mammals, including humans, is significantly larger than previously recognized, and frequent exposure through meat consumption and contact with animal or human feces provides ample opportunities for cyclovirus transmission. Determining the role of cycloviruses, found in 7 to 17% of non-U.S. human stools and 3 to 55% of non-U.S. meat samples tested, in both human and animal diseases is now facilitated by knowledge of their genomes.
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            Molecular evolution of porcine circovirus type 2 genomes: phylogeny and clonality.

            Porcine circoviruses (PCVs) type 1 (PCV1) and type 2 (PCV2) show high levels of nucleotide similarity, but PCV1 is considered non-pathogenic and PCV2 has been associated with several disease outcomes in pigs, mainly postweaning multisystemic wasting syndrome (PMWS). After exploring different topologies of the origin of PCVs, it was concluded that PCV1 and PCV2 seem to have a common origin. On the other hand, PCV2 could be divided into two groups (1 and 2) and eight clusters (1A to 1C and 2A to 2E), but none of those was apparently associated with disease status or geographic area. When phylogenetic trees constructed with the whole PCV2 genome, the cap or the rep genes were compared, some incongruence was identified. The possible existence of recombination was evaluated and cluster 1B was found to have a possible recombinant origin. Selective pressure was detected in all parts of the PCV2 genome, especially in the rep gene. Finally, the cap gene was the more suitable phylogenetic and epidemiological marker for PCV2, despite the fact that the virus can undergo recombination mainly within the first part of the rep region.
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              Genomic analysis of PCV2 isolates from Danish archives and a current PMWS case-control study supports a shift in genotypes with time.

              Porcine circovirus type 2 (PCV2) is the primary cause of Postweaning Multisystemic Wasting Syndrome (PMWS) in pigs. PCV2, however, is found in both PMWS-affected herds and non-affected herds. The objective of this study was to clarify if PCV2 genome nucleotide sequences isolated from pigs from PMWS-affected herds and non-affected herds cluster phylogenetically in two separate groups. All isolates (45) belonged to PCV2 group 1 and shared a nucleotide sequence identity of 99.4-100% indicating a very homogeneous PCV2 population in Denmark. Phylogenetic analysis of the PCV2 isolates revealed no distinctive clustering of case- and control-herds suggesting that there is no link between PCV2 sequences and herd disease status. The appearance of only PCV2 group 1 isolates in this study (isolates from 2003/2004) led us to determine if PCV2 nucleotide sequences had changed in Denmark over time. Interestingly, all PCV2 isolates from before the first outbreak of PMWS (2001) belonged either to a new PCV2 group identified for the first time in this study and named group 3 (isolates from 1980, 1987 and 1990) or PCV2 group 2 (isolates from 1993 and 1996). The shift from PCV2 group 2 to 1 was confirmed on a more global scale by placing all full genome PCV2 sequences submitted to GenBank from 1997 to 2006 in either of the groups by phylogenetic analysis. The analysis showed that the shift happened in 2003 or even earlier. This may indicate that PCV2 group 1 is a more adapted form of PCV2 and possibly could be more pathogenic.
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                Author and article information

                Journal
                Virus Evol
                Virus Evol
                vevolu
                Virus Evolution
                Oxford University Press
                2057-1577
                July 2019
                01 August 2019
                01 August 2019
                : 5
                : 2
                : vez026
                Affiliations
                Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, Merelbeke, Belgium
                Author notes
                Corresponding author: E-mail: hans.nauwynck@ 123456ugent.be
                Author information
                https://orcid.org/0000-0002-2648-3077
                https://orcid.org/0000-0002-3374-6605
                Article
                vez026
                10.1093/ve/vez026
                6676070
                31392030
                18098277-9060-43ee-bcd5-c0e66909d49a
                © The Author(s) 2019. Published by Oxford University Press.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

                History
                Page count
                Pages: 12
                Funding
                Funded by: China Scholarship Council 10.13039/501100004543
                Award ID: 201406300044
                Categories
                Research Article

                porcine circovirus type 2,phylogenetic analysis,genetic diversity,deduced amino acid sequence

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