Genomic characterization and high prevalence of bocaviruses in swine.
ABSTRACT Using random PCR amplification followed by plasmid subcloning and DNA sequencing, we detected bocavirus related sequences in 9 out of 17 porcine stool samples. Using primer walking, we sequenced the nearly complete genomes of two highly divergent bocaviruses we provisionally named porcine bocavirus 1 isolate H18 (PBoV1-H18) and porcine bocavirus 2 isolate A6 (PBoV2-A6) which differed by 51.8% in their NS1 protein. Phylogenetic analysis indicated that PBoV1-H18 was very closely related to a ∼2 Kb central region of a porcine bocavirus-like virus (PBo-LikeV) from Sweden described in 2009. PBoV2-A6 was very closely related to the porcine bocavirus genomes PBoV-1 and PBoV2 from China described in 2010. Among 340 fecal samples collected from different age, asymptomatic swine in five Chinese provinces, the prevalence of PBoV1-H18 and PBoV2-A6 related viruses were 45-75% and 55-70% respectively, with 30-47% of pigs co-infected. PBoV1-A6 related strains were highly conserved, while PBoV2-H18 related strains were more diverse, grouping into two genotypes corresponding to the previously described PBoV1 and PBoV2. Together with the recently described partial bocavirus genomes labeled V6 and V7, a total of three major porcine bocavirus clades have therefore been described to date. Further studies will be required to elucidate the possible pathogenic impact of these diverse bocaviruses either alone or in combination with other porcine viruses.
Article: Animal bocaviruses: a brief review.[show abstract] [hide abstract]
ABSTRACT: The recently discovered human Bocavirus has been preliminary classified into the genus Bocavirus of the family Parvoviridae. Animal bocaviruses have been known in veterinary medicine since the early 1960s. This paper reviews the current knowledge about the two eponymous members of the genus: bovine parvovirus (BPV) and canine minute virus (CnMV). In contrast to other parvoviruses, bocaviruses contain a third open reading frame in the mid of the genome encoding for a highly phosphorylated non-structural protein, NP1, whose function has not yet been determined. The VP1-unique region of BPV and CnMV contains a phospholipase A(2) sequence motif. Both viruses cause diseases of the gastrointestinal and respiratory tract and are known to infect fetuses and to cause reproductive disorders. Especially young animals suffer from disease, whereas in adults subclinical infection is common. Clinical signs include diarrhea, vomiting, dyspnea, embryonic/fetal death after transplacental infection early and abortion late in gestation. Both viruses have in common that they are widespread in their host species as worldwide serosurveys indicate. For BPV it has been shown that sialated glycoproteins mediate hemagglutination reaction and function as receptor for virus attachment on permissive cells.Intervirology 12/2008; 51(5):328-34. · 1.89 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: We determined the complete nucleotide sequence of bovine parvovirus (BPV), an autonomous parvovirus. The sequence is 5,491 nucleotides long. The terminal regions contain nonidentical imperfect palindromic sequences of 150 and 121 nucleotides. In the plus strand, there are three large open reading frames (left ORF, mid ORF, and right ORF) with coding capacities of 729, 255, and 685 amino acids, respectively. As with all parvoviruses studied to date, the left ORF of BPV codes for the nonstructural protein NS-1 and the right ORF codes for the major parts of the three capsid proteins. The mid ORF probably encodes the major part of the nonstructural protein NP-1. There are promoterlike sequences at map units 4.5, 12.8, and 38.7 and polyadenylation signals at map units 61.6, 64.6, and 98.5. BPV has little DNA homology with the defective parvovirus AAV, with the human autonomous parvovirus B19, or with the other autonomous parvoviruses sequenced (canine parvovirus, feline panleukopenia virus, H-1, and minute virus of mice). Even though the overall DNA homology of BPV with other parvoviruses is low, several small regions of high homology are observed when the amino acid sequences encoded by the left and right ORFs are compared. From these comparisons, it can be shown that the evolutionary relationship among the parvoviruses is B19 in equilibrium with AAV in equilibrium with BPV in equilibrium with MVM. The highly conserved amino acid sequences observed among all parvoviruses may be useful in the identification and detection of parvoviruses and in the design of a general parvovirus vaccine.Journal of Virology 01/1987; 60(3):1085-97. · 5.08 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: Four antigenically related transmissible agents were recovered from canine fecal specimens. The agents produced cytopathic effects in a continuous dog cell line developed in this laboratory. Increased antibody titers were demonstrated in three of the four dogs which provided the isolates. The virus did not produce cytopathic effects in primary canine kidney or thymus cell cultures, or in cell cultures of human, simian, porcine, bovine, feline, and murine origin. The agent is resistant to ether, chloroform, and heat treatment, and the growth of the virus is inhibited by 5-iodo-2-deoxyuridine. After acridine orange staining, green fluorescent intranuclear inclusions are seen in infected cell cultures. By electron microscopy, the virions measure approximately 20 to 21 nm in overall diameter and are present in the nuclei of infected cells. These properties are consistent with membership in the parvovirus or picodnavirus group. The agent hemagglutinates rhesus red blood cells at 5 C and by hemagglutination-inhibition tests could be readily distinguished from H-1, rat virus, and the minute virus of mice. Canine gamma globulin contains high titers of neutralizing antibody and neutralizing antibody was found in a high percentage of military dogs and in beagles of a breeding colony.Infection and Immunity 06/1970; 1(5):503-8. · 4.07 Impact Factor
Genomic Characterization and High Prevalence of
Bocaviruses in Swine
Tongling Shan1,2,3., Daoliang Lan1., Linlin Li2,3, Chunmei Wang4, Li Cui1, Wen Zhang5, Xiuguo Hua1*,
Caixia Zhu1, Wei Zhao1, Eric Delwart2,3
1Zoonosis and Comparative Medicine Group, Shanghai Jiao Tong University, Shanghai, People’s Republic of China, 2Blood Systems Research Institute, San Francisco,
California, United States of America, 3Department of Laboratory Medicine, University of California San Francisco, San Francisco, California, United States of America,
4College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Huibei, People’s Republic of China, 5School of Medical Science and Laboratory Medicine,
Jiangsu University, Zhenjiang, Jiangsu, People’s Republic of China
Using random PCR amplification followed by plasmid subcloning and DNA sequencing, we detected bocavirus related
sequences in 9 out of 17 porcine stool samples. Using primer walking, we sequenced the nearly complete genomes of two
highly divergent bocaviruses we provisionally named porcine bocavirus 1 isolate H18 (PBoV1-H18) and porcine bocavirus 2
isolate A6 (PBoV2-A6) which differed by 51.8% in their NS1 protein. Phylogenetic analysis indicated that PBoV1-H18 was
very closely related to a ,2 Kb central region of a porcine bocavirus-like virus (PBo-LikeV) from Sweden described in 2009.
PBoV2-A6 was very closely related to the porcine bocavirus genomes PBoV-1 and PBoV2 from China described in 2010.
Among 340 fecal samples collected from different age, asymptomatic swine in five Chinese provinces, the prevalence of
PBoV1-H18 and PBoV2-A6 related viruses were 45–75% and 55–70% respectively, with 30–47% of pigs co-infected. PBoV1-
A6 related strains were highly conserved, while PBoV2-H18 related strains were more diverse, grouping into two genotypes
corresponding to the previously described PBoV1 and PBoV2. Together with the recently described partial bocavirus
genomes labeled V6 and V7, a total of three major porcine bocavirus clades have therefore been described to date. Further
studies will be required to elucidate the possible pathogenic impact of these diverse bocaviruses either alone or in
combination with other porcine viruses.
Citation: Shan T, Lan D, Li L, Wang C, Cui L, et al. (2011) Genomic Characterization and High Prevalence of Bocaviruses in Swine. PLoS ONE 6(4): e17292.
Editor: Daniel G. Bausch, Tulane School of Public Health and Tropical Medicine, United States of America
Received August 24, 2010; Accepted January 27, 2011; Published April 15, 2011
Copyright: ? 2011 Shan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by Key Project of Shanghai Science and Technology Committee of China under Grant No. 063919121. The funders had no
role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests: The authors have declared that no competing interests exist.
* E-mail: email@example.com
. These authors contributed equally to this work.
Members of the Parvoviridae family are small non-lipid enveloped
viruses with a diameter of 18–26 nm, icosahedral symmetry
(T=1), encoded by a single-stranded linear DNA genome of
approximately 4,000 to 6,000 nucleotides (nt) . The family
includes two subfamilies: Densovirinae, Parvovirinae. The subfamily of
Densovirinae contains four genera: Densovirus, Iteravirus, Brevidensovirus
and Pefudensovirus, which infect only invertebrates . The
Parvovirinae subfamily is currently subdivided into five genera:
Parvovirus, Erythrovirus, Dependovirus (adeno-associated virus), Amdovirus,
and Bocavirus, infecting vertebrates .
The bocavirus genus was recently assigned by the International
Committee on Taxonomy of Viruses (ICTV)  to parvovirus
genomes containing a third ORF (labeled NP1) between the NS1
and VP1/VP2 genes . Bocaviruses were first identified in
bovine and canine [3,4], samples from which it derives its genus
name [1,5]. Presently, the bocavirus genus contains eight
members: bovine parvovirus, canine minute virus (CnMV), human
bocavirus 1–4 (HBoV1-4), a gorilla bocavirus and a partially
sequenced chimpanzee bocavirus [1,6,7].
The first human bocavirus (HBoV) was found in the
nasopharyngeal secretion of a child with respiratory problems
using a methodology closely related to that used here . HBoV
has been associated with lower respiratory tract symptoms and
possibly diarrhea [5,9–22], and shows a very low degree of genetic
variability worldwide [6,23,24]. HBoV2 was first reported in the
stool of Pakistani children with non-polio acute flaccid paralysis
(AFP) , and then in Australian children and Chinese children
with diarrhea [9,25]. HBoV3 was first reported in the stool of
Australian children with diarrhea  and then in stool from
Nigerian, Tunisian, Nepalese and US children . HBoV4 was
reported in the stool of children with AFP from Nigeria and
Tunisia . HBoV 1/2/3 were also detected in untreated sewage
water from throughout the US . Recently, a novel bocavirus
was identified in the feces of captive gorillas with diarrhea  and
from wild gorillas and chimpanzees . In 2009, a porcine boca-
like virus (PBo-likeV) was reported in swine feces with postweaning
multisystemic wasting syndrome in Sweden and 1854 bp of its
partial genome sequenced . In 2010, the nearly complete
genomes of distinct porcine bocaviruses provisionally named
PBoV1 and PBoV2 were characterized from feces of swine in
China . Finally, partial genome sequences of 2.4 Kb from
another clade of porcine bocaviruses labeled 6V and 7V where
also identified yielding three major bocavirus groups in swine
(PBo-likeV, PBoV1/PBoV2, and 6V/7V).
PLoS ONE | www.plosone.org1 April 2011 | Volume 6 | Issue 4 | e17292
Random amplification and sequencing has been used to
discover novel virus in human and animal [8,10,29–53]. In this
study, we found in swine feces highly distinct bocavirus whose
genome we tentatively named porcine bocavirus 1 (PBoV1-H18),
and porcine bocavirus 2 (PBoV 2-A6). The nearly complete
genomes of both viruses were acquired and are described here.
PBoV1-H18 and PBoV2-A6 were also screened for in 340 stool
samples of asymptomatic swine from five provinces of China.
Materials and Methods
A total of 340 porcine stool samples from different aged swine
were collected from 17 middle or large-scale porcine farms (200–
2,000 sows each) in five provinces of China from April 2008 to
October 2009, of which 80 were collected from four farms located
in Shanghai, 60 from three farms located in Jiangsu province, 120
from six farms located in Anhui province, 20 from a farm located
in Shandong province, and 60 from three farms located in
Guizhou province and stored at 280uC (Table 1).
Viral particle purification and RT-PCR
One stool sample was randomly selected from each of the 17
farms. The samples were suspended in PBS (0.01 M phosphate,
pH7.2–7.4, 0.15NaCl), vortexed, centrifuged at 15000 g, and
filtered through a 0.22-mm filter to remove eukaryotic- and
bacterial-cell-sized particles [45,46,54,55]. The filtrates were then
treated with benzonase, DNase and RNase to digest non-particle-
protected nucleic acid as reported [54,55]. Viral nucleic acids were
then extracted using the TIANamp virus DNA/RNA Kit
(TIANGEN BIOTECH, BEIJING, CO., Ltd.). Viral cDNA
synthesis was performed by incubation of the extracted viral
RNA/DNA with 100 pmol of primer K-8N  with a
degenerate 39 end and the use of Superscript reverse transcriptase,
and the opposite strand of the cDNA was generated after melting
and reannealing and primer extension using Klenow DNA
polymerase [45,46,54–56]. PCR of extension products was
performed as reported previously using the K-8 primer (K-8N
without the degenerate 39 end) . This protocol amplifies both
viral RNA and DNA genomes [54,55].
Novel virus identification and complete genome
Random RT-PCR DNA products ran as smears on agarose gel
and were gel purified (Axygen, CA, USA), then subcloned into
pMD-18T plasmid vector (TaKaRa, Japan) for sequencing. The
sequences were then screened for sequence similarities using
tBLASTx and BLASTn against the nr database in GenBank.
DNA and RNA were also directly extracted from centrifuged
stool supernatant using the TIANamp virus DNA/RNA Kit
(TIANGEN BIOTECH, BEIJING, CO., Ltd.). For PBoV1-H18
related sequences screening, primers PBoV1-L1 (59-CTGTGG-
TGTGTATTTGTGG-39) were used for the first round of nested
PCR, and the primers PBoV1-L2 (59-ACAGGAATTAACAGAC-
GAAG-39) and PBoV1-R2 (59-TATCGGCACGTACCATT-
GAC-39) were used for the second round of nested PCR, resulting
in the amplification of a 530-bp fragment over the NP1 and VP1
genes. PBoV2-A6 related sequences were detected using primers
PBoV2-L1 (59-CAAGGGCGCTACACACACAA-39) and PBoV2-
R1 (59-TTAATTCCGCACTTAGTTGG-39) for the first round of
nested PCR; primers PBoV2-L2 (59-TCCAGTAACCAAAACA-
TACC-39) and PBoV2-R2 (59- TCTCGTGTTGATTGTAG-
CTC-39) for the second round of nested PCR, amplifying a 439-
bp fragment of the VP1 gene.
Sequence and phylogenetic analysis
Sequences of each PCR product were assembled using SeqMan
II program (DNASTAR, Inc). The identification of open reading
Table 1. High prevalence of PBoV1-H18 and PBoV2-A6 detected by nested–PCR assay in fecal samples from different aged pigs in
five provinces of China.
(province)Number of farms
Positive/tested samples for
Positive/tested samples for
49/80 (61.3%)47/80 (58.8%) 31/80 (38.8%)
35/60 (58.3%) 41/60 (68.3%)22/60 (36.7%)
77/120 (64.2%)81/120 (70.1%) 46/120 (38.3%)
9/20 (45%) 11/20 (55%)6/20 (30%)
45/60 (75%)39/60 (65%)28/60 (46.7%)
Total 17 215/340 (63.2%)219/340 (64.4%)133/340 (39.1%)
A New Bocavirus Species in Swine
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frames (ORFs) was performed by a translated BLAST search
(BLASTx at http://www.ncbi.nlm.nih.gov/blast/Blast.cgi) and an
ORF finder at the website (http://www.ncbi.nlm.nih.gov/gorf/
gorf.html). Sequences used for the comparison were comprised the
GQ867667, FJ973561, FJ375129, FJ170278 and NC_012042),
GBoV1 (HM145750), CnMV (AB158475, FJ899734, AF495467
and FJ214110), BPV1 (DQ335247 and NC_001540), canine
parvovirus (D26079, NC_001539 and EU310373), porcine
parvovirus (EU790642), mice minute virus (J02275), mouse
porcine parvovirus 4 (HM031135, GQ387500, GQ387499),
human parvovirus B19 (FJ591158), simian parvovirus (U26342),
human parvovirus 4 (AY622943), goose parvovirus (NC_001701),
Muscovy duck parvovirus (NC_006147), PBo-likeV (FJ872544),
PBoV1 and PBoV2 (HM053693 and HM053694), and 6V and 7V
(HM053672 and HM053673). Multiple sequence alignment was
performed using CLUSTAL W. Protein amino acid distances were
calculated using the MegAlign program (DNASTAR, Inc).
Phylogenetic trees were generated using the neighbor joining
(NJ) method with bootstrap of 1,000 replicates with MEGA 4.1
Nucleotide sequence accession numbers
The near-full genomes of PBoV1-H18, PBoV2-A6 and the
partial NS1 and VP1 sequences from the diagnostic nPCR have
been deposited in GenBank under accession numbers HQ291308-
HQ291309 and HQ291310-HQ291343.
Novel porcine bocavirus sequence
Seventeen porcine samples stool supernatants from 17 farms
were analyzed using a generic viral particle-protected nucleic acid
enrichment procedure followed by random amplification of
extracted RNA and DNA (see materials and methods) [46,54–
56]. Amplified DNA was then subcloned and 1190 plasmid inserts
were sequenced (70 for each of 17 samples). Nine samples (totaling
82 subclones) showed the presence of fragments whose virtual
translation products were related to canine and human boca-
viruses using BLASTx. Twenty-four clones from pig sample H18
and 26 clones from pig sample A6 showed significant similarity
with bocaviruses. H18 derived sequences showed high (.99%)
identity with the recently described porcine bocavirus-like virus
(PBo-LikeV), the only porcine bocavirus reported at the time of
these experiments (GenBank GU902971) . Sequences from
porcine sample A6 showed low identity with those of H18 and
PBo-likeV. The H18 and A6 samples were selected for targeted
viral genome amplification and sequencing.
Nearly complete genomes of PBoV1 and PBoV2
The 24 sequences from H18 were assembled to form a
continuous sequence of approximately 2700 nucleotides that
appeared to lack .1300 and 1000 nucleotides from the 59and 39
ends of its genome. PCR primers based on the available H18
sequences and regions highly conserved between HBoV2
(FJ170278) and canine bocavirus (FJ214110) were used to amplify
the nearly complete bocavirus genome we provisionally called
PBoV1-H18 (5267 nt). To confirm this genome sequence, this
sequence was re-amplified using 6 sets of PCR primers generating
overlapping fragments of the genome which were directly
sequenced. Using the same method, the nearly complete bocavirus
genome (5117 nt) from sample A6 was also sequenced and was
provisionally labeled PBoV2-A6.
Using an open reading frame (ORF) finder (http://www.ncbi.
nlm.nih.gov/gorf/gorf.html), three ORF were found in both
genomes (Figure 1). The ORFs of PBoV1-H18 were 636 aa for
NS1, 219 aa for NP1 and 621 aa for VP1/VP2. The ORFs of
PBoV2-A6 were 703 aa for NS1, 221 aa for NP1 and 704 aa for
VP1/VP2. The possible splicing of bocavirus NS1 transcripts
recently shown to extend the length of NS1 proteins was not
investigated here [6,57].
Figure 1. The PBoV1 and PBoV2 genomes. Diagrammatic representation of PBoV1-H18 and PBoV2-A6 sequences showing the position of ORFs
for NS1, NP1, VP1 and VP2, compared with other bocaviruses. The black boxes represent the known spliced exon of NS1 transcripts.
A New Bocavirus Species in Swine
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A New Bocavirus Species in Swine
PLoS ONE | www.plosone.org4April 2011 | Volume 6 | Issue 4 | e17292
Prevalence of PBoV1 and PBoV2
Nucleic acids were extracted from 340 porcine stool samples.
PBoV1-H18 related sequences were screened for using nested
PCR with primers amplifying a 530-bp fragment of the NP1/VP1
region. The electrophoretic bands of the expected size were
subcloned and sequenced. The results showed that the prevalence
of PBoV1-H18 related viruses was high in China with 215 out of
340 (63.2%) porcine samples positive (Table 1). All PBoV1-H18
related sequences showed .99% identity with each other. For
PBoV2-A6 nested PCR, 219 out of 340 (64.4%) samples were
positive (Table 1), with the amplicons showing 90% to 100%
identity with each other. 133 out of 340 (39.1%) samples were co-
infected with both PBoV1-H18 and PBoV2-A6 related viruses
Phylogenetic analysis of PBoV1 and PBoV2
To determine the genetic relationship of PBoV1-H18 and
PBoV2-A6 with recently described porcine bocaciruses and
bocaviruses from other host species, both nucleotide and amino
acid alignments were generated and used for phylogenetic
analyses. When the whole genomes were considered porcine
bocaviruses as a group (except for the V6/V7 variants with only
NP1 sequences available), were most closely related to the canine
bocavirus CnMV (Figure 2A). Phylogenetic analyses of the 3
ORFs – NS1, NP1 and VP1/VP2 – were also performed
(Figure 2B–G). In all three regions, PBoV1-H18 was most closely
related to the Chinese PBoV1 and PBoV2 recently reported by
Cheng et al . PBoV2-A6 was closely related in NP1 to the first
reported partial porcine bocavirus sequence PBo-likeV from
Sweden, the only region available for comparison (Fig. 2C, F)
. Table 2 numerically shows the protein similarities between
PBoV1-H18 and PBoV2-A6 and other porcine and non-porcine
The partial VP1 sequence of PBoV2-A6 related variants from
different farms was also phylogentically analyzed and fell into two
major clades we named PBoV2 genotype 1 and 2 (PBoV2-G1 and
PBoV2-G2). The two previously described Chinese PBoV1 and
PBoV2 ‘‘species’’ grouped within these two genotypes.
PBo-likeV was originally found in swine with postweaning
multisystemic wasting syndrome (PMWS) in 2009 when approx-
imately 35% of its genome sequence was reported . The nearly
full genomes of two distantly related porcine bocaviruses labeled
PBoV1 and PBoV2 as well as two partial genomes labeled V6 and
V7 were then reported in 2010 . These bocaviruses grouped
into 3 phylogenetic clades containing PBo-likeV, PBoV1/PBoV2,
and V6/V7. In the present study, we characterize two nearly
complete bocavirus genomes, one of which (PBoV1-H18) grouped
with the PBo-likeV clade while the second (PBoV2-A6) fell with
the PBoV1/PBoV2 clade. We provisionally named the genome
from the H18 sample PBoV1-H18 since its closest homologue,
PBo-likeV, was the first reported porcine bocavirus . The virus
from sample A6 was provisionally named PBoV2-A6 since it
phylogenetically groups with the second reported set of porcine
bocaviruses containing both PBoV1 and PBoV2 . No close
homologues of the V6/V7 clade were identified in this study.
Under this proposed classification scheme, the viruses labeled
PBoV1 and PBoV2 by Cheng et al, therefore both belong to the
PBoV2 clade, the second reported clade of porcine bocaviruses
. Under this proposed taxonomic classification, the V6 and V7
bocaviruses  belong to the still only partially characterized
Sequence analysis of the PBoV2 clade showed that their VP1/
VP2 genes were highly diverse and could be classified into two
genotypes (Figure 3). The partial NP1 and VP1 genes of PBoV1-
H18 related viruses detected in this study were more highly
conserved (99–100% identity), consistent with a recent report by
Zhai et al reporting PBoV1 in Chinese pigs using partial VP1/
VP2 nested PCR and sequencing . Zhai et al also found a high
prevalence of PBoV1 (69% in weanling piglets) with a higher
frequency of PBoV1 in animals also infected with PCV2, PRRSV,
PTTV or CSFV and in pigs with respiratory symptoms versus
healthy pigs .
Members of the bocavirus genus contain an 3rdORF of
unknown function labeled NP1 gene. Recently another parvovirus
(PPV4) was identified in porcine feces that also contained a central
Figure 2. Phylogenetic tree constructed by the neighbor-joining method with 1,000 bootstrap replicates using MEGA4.0 software.
Bootstrap values are indicated at each branching point. Scale bar indicates estimated genetic distance. Phylogenetic analysis of the nearly full-length
genomes of PBoV1 and PBoV2 and 33 representative bocavirus species/strains (A). Phylogenetic analysis of nucleic acid and amino acid of NS1 (B and
E), NP1 (C and F) and VP1 (D and G) ORFs of PBoV1-H18 and PBoV2-A6 and representative bocavirus species.
Table 2. Comparison of pairwise amino acid distances (p-distance) of three genes between PBoV1-H18, PBoV2-A6 and other
NS1- 51.8-52.9 51.7-- 59.6 58.659.4 59.2 59.755.865.8
NP1- 62.30 64.262.2-- 67.967.668.766.2 68.8 64.8 67.6
VP1-52.3- 52.052.7 63.163.4 61.862.8 62.4 62.762.353.663.2
NS151.8--6.8 5.8-- 58.758.3 60.158.362.2 55.966.7
NP162.3- 62.37.9 10.1-- 54.758.155.1 56.757.551.9 58.1
VP152.3--2.7 7.159.259.552.1 53.153.752.753.546.955.0
A New Bocavirus Species in Swine
PLoS ONE | www.plosone.org5April 2011 | Volume 6 | Issue 4 | e17292
3rdORF, although unrelated in sequence to the bocaviruses NP1
. None of the ORFs of PPV4 clustered phylogenetically with
the bocaviruses (data not shown) but instead clustered with
members of the parvovirus genus . PPV4 is therefore
unrelated to the bocaviruses reported here.
HBoV1 has been associated with respiratory symptoms while
other HBoV may be associated with diarrhea and acute flaccid
paralysis [5,8–10,11–22,25]. A gorilla bocavirus was detected in
captive animals in the US experiencing severe diarrhea .
PBoV1 was found in pigs with PMWS in Sweden  and in pigs
with respiratory tract symptoms in China . In the present
study, both PBoV1 and PBoV2 were highly prevalent in both
asymptomatic swine from five provinces of China. Further studies
are needed to examine possible associations between infections
with these different porcine bocaviruses, the viral loads excreted,
the presence of co-infections and various porcine diseases.
Conceived and designed the experiments: TS DL CW XH ED. Performed
the experiments: TS DL CW LC CZ WZ. Analyzed the data: TS DL CW
LL CZ WZ ED. Contributed reagents/materials/analysis tools: TS DL
CW LL LC CZ WZ XH ED. Wrote the paper: TS.
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