The phylogenetic trees were constructed by using the Maximum Likelihood method based on the Poisson correction model, with the amino acid sequences of the nonstructural protein (ORF1, NS1) and the capsid protein (ORF2, VP1).

<p>The percentage of the tree in which the associated sequences clustered together is shown above the branches (only >70% are shown). The trees are drawn to scale, with branch lengths measured in the number of substitutions per site. Evolutionary analyses were conducted in MEGA5 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0065312#pone.0065312-Tamura1" target="_blank">[34]</a>. The analyses involved 39 sequences of vertebrate parvoviruses with their GenBank accession numbers marked in the tree. PPV5 sequences characterized in the present study are in bold and marked with “♦”. The sequence of <i>Mythimna loreyi</i> densovirus (AY461507) was used as outgroup to root the tree. The asterisk indicates the new genus proposed by ICTV. (A) Tree constructed based on the amino acid sequences of NS1. All positions containing gaps and missing data except ambiguous positions were included. There were a total of 900 positions in the final dataset. (B) Tree constructed based on the amino acid sequences of VP1. All positions containing gaps and missing data except ambiguous positions were included. There were a total of 1,177 positions in the final dataset.</p

Summary of the history (age, states, farms) and prevalence of PPV4, PPV5 or concurrent PPV4/5 DNA in 185 fecal samples from pigs in the U.S.

<p>Summary of the history (age, states, farms) and prevalence of PPV4, PPV5 or concurrent PPV4/5 DNA in 185 fecal samples from pigs in the U.S.</p

The phylogenetic trees were constructed by using the Maximum Likelihood method based on the Kimura 2-parameter model, with the nucleotide sequences the nonstructural protein (ORF1, NS1) and the capsid protein (ORF2, VP1).

<p>The percentage of the tree in which the associated sequences clustered together is shown above the branches (only those >70% are shown). The trees are drawn to scale, with branch lengths measured in the number of substitutions per site. Evolutionary analyses were conducted in MEGA5. The analyses involved 39 sequences of vertebrate parvovirus with their GenBank accession numbers marked in the tree. PPV5 sequences characterized in the present study are in bold and marked with “♦”. The sequence of <i>Mythimna loreyi</i> densovirus (AY461507) is used as outgroup to root the tree. The asterisk indicates the new genus proposed by ICTV. (A) Tree constructed based on the nucleotide sequence of NS1. All positions containing gaps and missing data except ambiguous positions were included. There were a total of 2,690 positions in the final dataset. (B) Tree constructed based on the nucleotide sequence of VP1. All positions containing gaps and missing data except ambiguous positions were included. There were a total of 3,386 positions in the final dataset.</p

Sequence alignment of the putative phospholipase A₂ motif of PPV5 with those of other PPVs, PBoV, and BPV2.

<p>The Ca²⁺ binding loop is indicated by filled squares and the catalytic residues by filled circles. The positions of the amino acids and the GenBank numbers of the sequences are indicated.</p

Primers and probes used for PPV4 and PPV5 detection.

a<p>Nucleotide position according to PPV4 (GenBank accession no. GQ387500).</p>b<p>Nucleotide position according to PPV5 (GenBank accession no. JX896321).</p

Primers used for the PPV5 genome sequencing.

<p>Primers used for the PPV5 genome sequencing.</p

Schematic representation of the genomes of PPV5 compared with PPV4 and BPV2.

<p>The position and size of putative ORFs are indicated. The GenBank accession numbers of the reference sequences are JX896321 (PPV5), GQ387499 (PPV4), AF406966 (BPV2). aa: amino acid. NS: nonstructural gene.</p

ASFV recombinant p30 antibody ELISA serum-to-positive (S/P) responses in serum (mean, SE) and oral fluid (mean, SE) samples from 17 pigs (experiments 1 and 2) inoculated with ASFV NHV/P68.

<p>ASFV recombinant p30 antibody ELISA serum-to-positive (S/P) responses in serum (mean, SE) and oral fluid (mean, SE) samples from 17 pigs (experiments 1 and 2) inoculated with ASFV NHV/P68.</p

African swine fever virus (ASFV) antigen-specific serum antibody responses detected by multiplex fluorescent microbead-based immunoassay (FMIA) in pigs inoculated with replicon particle (RP) vaccines expressing p30, p54, and/or p72 genes.

<p>African swine fever virus (ASFV) antigen-specific serum antibody responses detected by multiplex fluorescent microbead-based immunoassay (FMIA) in pigs inoculated with replicon particle (RP) vaccines expressing p30, p54, and/or p72 genes.</p

ASFV multiplex fluorescent microbead-based immunoassay (FMIA) sample-to-positive (S/P) serum antibody response (mean, SE) against three recombinant antigens (p30, p54, p72) in 9 pigs inoculated with ASFV NHV/P68 (experiment 1).

<p>ASFV multiplex fluorescent microbead-based immunoassay (FMIA) sample-to-positive (S/P) serum antibody response (mean, SE) against three recombinant antigens (p30, p54, p72) in 9 pigs inoculated with ASFV NHV/P68 (experiment 1).</p