A porcine parvovirus, designated as VRI-1, was isolated from a 30-day-old piglet. Replicative form of viral DNA from ST cells infected with VRI-1 was directly cloned into pUC19. The cloned DNA fragment contained the entire nonstructural and structural protein genes, covering approximately 85% of the viral genome. The nucleotide sequence of VRI-1 showed 99.4~99.5% identity in the nonstructural protein (NS) and 99.0~99.2% identity in the structural protein with previously reported PPV strains, respectively. Among the cloned genes, two types of defective genomes with deletion of 100 and 247 nucleotides at almost similar location of 3' region within NS gene were also identified in this study.
Base Sequence* ; Clone Cells ; DNA ; DNA, Viral ; Genome* ; Genome, Viral ; Nucleotides ; Parvovirus, Porcine*

Porcine parvovirus, Erysipelothrix (E.) rhusiopathiae, and Leptospira (L.) interrogans are considered major etiologic agents of reproductive failure in pigs, causing economic loss in the swine industry. In this study, the safety and immunogenicity of a new octavalent inactivated vaccine were evaluated. The vaccine contained inactivated porcine parvovirus, E. rhusiopathiae, and six L. interrogans serovars (Bratislava, Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, and Pomona). Safety test results showed no notable side effects or clinical signs after vaccination in mice, guinea pigs, and sows. In addition, we assessed immunogenicity of the vaccine in 25 sows under field conditions. The vaccinated group (n = 20) had a significantly higher antibody level than the non-vaccinated group (n = 5). Moreover, the stillbirth rate decreased in piglets born from vaccinated sows, resulting in an increased fertility rate. The results of this study demonstrate that the new octavalent inactivated vaccine can be applied safely and effectively to improve reproductive performance in sows.
Animals ; Birth Rate ; Erysipelas* ; Erysipelothrix ; Guinea Pigs ; Leptospira* ; Leptospirosis ; Mice ; Parvovirus, Porcine* ; Serogroup ; Stillbirth ; Swine ; Vaccination

Postweaning multisystemic wasting syndrome (PMWS), which was first identified in western Canada in 1991 and more recently in the United States, Europe and Asia, is an emerging disease in pigs. Porcine circovirus type 2 (PCV-2) is the primary infectious viral agent causing PMWS, but the full expression of the disease may require the presence of other agents. It is reported that there is apparent synergism between PCV-2 and porcine parvovirus (PPV) in increasing the severity of the clinical signs and lesions of PMWS. From January 2006 to May 2008, a total of the 154 lymph node samples were collected from 4~12 weeks old pigs which had been submitted to the College of Veterinary Medicine, Jeju National University, Korea. These pigs were diagnosed as PMWS on the basis of clinical and pathological examination from 48 commercial herds in Jeju Island. Based on the immunohistochemistry, porcine parvovirus was detected in 69 cases (44.8%) from 154 weaned or grower pigs. PPV antigens were detected in the cytoplasm of histiocytic cells multifocally infiltrated in the cortex and paracortex of lymph nodes. The results of this study clarify that PPV is prevalent in pigs with PMWS on Jeju Island. Therefore PPV is one of the most important co-agents in the development of naturally acquired PMWS. This study may be helpful to the control of this disease and to epidemiological aspects.
Asia ; Canada ; Circovirus ; Coinfection ; Cytoplasm ; Europe ; Immunohistochemistry ; Korea ; Lymph Nodes ; Parvovirus, Porcine ; Prevalence ; Swine ; United States ; Veterinary Medicine ; Wasting Syndrome

Porcine parvovirus 7 (PPV7) was first detected in Korean pig farms in 2017. The detection rate of PPV7 DNA was 24.0% (30/125) in aborted pig fetuses and 74.9% (262/350) in finishing pigs, suggesting that PPV7 has circulated among Korean domestic pig farms. Phylogenetic analysis based on capsid protein amino acid sequences demonstrated that the nine isolated Korean strains (PPV-KA1-3 and PPV-KF1-6) were closely related to the previously reported USA and Chinese PPV7 strains. In addition, the Korean strains exhibit genetic diversity with both insertion and deletion mutations. This study contributes to the understanding of the molecular epidemiology of PPV7 in Korea.
Aborted Fetus ; Agriculture ; Amino Acid Sequence ; Asian Continental Ancestry Group ; Capsid Proteins ; DNA ; Fetus ; Genetic Variation ; Humans ; Korea ; Molecular Epidemiology ; Parvovirus, Porcine ; Sequence Deletion ; Sus scrofa ; Swine

To study the proliferation characteristics of PPV in differently infected way and the variance of concentrations in different cells. A strain of porcine parvovirus(PPV) was adapted to PK-15 cells, and a Real-time fluorescent quantitative PCR (FQ-PCR) assay was developed based on the specific region of the NS1 gene of PPV to quantify the PPV. The FQ-PCR was used to measure the viral concentration of virus-infected cells by simultaneous or step by step inoculation and plot one-step growth curves. The proliferation characteristics of PPV strain in different cells lines (HeLa, MDBK, PK-15 ,ST, F81, BHK-21 and Marc-145) was also compared. The results showed the PK-15 cell -adapted strain of PPV produced CPE after 12 passages, and maintained stable CPE at the following 10 messages. The one-step growth curve showed that the virus concentration of simultaneous inoculation was higher than that of the step-by-step inoculation, and the proliferation cycle of step-by-step inoculation was shorter. The proliferation ability of PPV strain in different cells showed that CPE appeared first inPK-15, followed by ST, HeLa and MDBK, and the virus concentration was highest in ST, followed byPK-15, MDBK and HeLa. NO proliferation was observed in F81, BHK-21 and Marc-145 cells. These findings lay a material foundation for the basic researches on PPV and the development of vaccine.
Animals ; Cell Line ; Cricetinae ; Cytopathogenic Effect, Viral ; DNA, Viral ; genetics ; Female ; Haplorhini ; Humans ; Male ; Parvoviridae Infections ; virology ; Parvovirus, Porcine ; genetics ; physiology ; Real-Time Polymerase Chain Reaction ; Reproducibility of Results ; Sensitivity and Specificity ; Swine ; Viral Proteins ; genetics ; Virus Replication

Porcine reproductive and respiratory syndrome virus(PRRSV)0, porcine circovirus type 2(PCV-2) and porcine parvovirus (PPV)0 infections were investigated as possible causes of the postweaning multisystemic wasting syndrome(PMWS). Specific primers for RT-PCR and PCR were designed for the differential detection of PRRSV, PCV-2 and PPV. Using PCR, these viruses were detected in homogenized tissue samples from pigs that had respiratory of reproductive problems in the time period between 1998 and 2000; the overall prevalences were: PRRSV 31.4%, PCV-2 46.5%, and PPV 8.1%. PCV-2 was also detected in aborted fetal tissues.
Aborted Fetus/virology ; Animals ; Base Sequence ; Circoviridae Infections/diagnosis/epidemiology/*veterinary ; Circovirus/genetics/isolation&purification ; DNA Primers ; Diagnosis, Differential ; Korea/epidemiology ; Parvoviridae Infections/diagnosis/epidemiology/*veterinary ; Parvovirus, Porcine/genetics/isolation&purification ; Polymerase Chain Reaction/methods/veterinary ; Porcine Reproductive and Respiratory Syndrome/diagnosis/*epidemiology ; Porcine respiratory and reproductive syndrome virus/genetics/isolation & purification ; Prevalence ; Respiratory Tract Infections/veterinary/virology ; Reverse Transcriptase Polymerase Chain Reaction/methods/veterinary ; Sequence Homology ; Swine ; Swine Diseases/diagnosis/*epidemiology ; Wasting Syndrome/*veterinary/virology

Rats were administered zearalenone (ZEA) via gavage at dosages of 0, 1, 5, and 30 mg/kg for 36 days. On treatment day 8, inactivated porcine parvovirus vaccine (Vac) was injected intraperitoneally. Antibody production against porcine parvovirus was then measured as a function of ZEA treatment. Compared to the vaccine alone, ZEA treatment, with or without Vac, decreased the serum level of IgG. The level of IgM decreased in all ZEA groups at day 22, but the decrease was sustained only in the medium-dose ZEA group at day 36. The level of IgA was unchanged in the Vac only and ZEA groups at day 22, but was decreased in the 5 mg/kg ZEA plus Vac group compared to the Vac only group at day 36. The level of IgE was decreased by all doses of ZEA at day 22, but was unaffected in ZEA plus Vac groups compared to the Vac only group. The levels of IL-1 in the thymus and spleen; INF-gamma in serum; IL-2, IL-6, and IL-10 in the thymus; and IL-10 and IFN-gamma in the spleen decreased after ZEA administration. Furthermore, the levels of IL-1beta in the spleen and mesenteric lymph node, IL-1beta in the thymus, IL-2 in the thymus and spleen, IL-6 in the thymus, IL-10 and IFN-gamma in the spleen, and GM-CSF and TNF-alpha in the thymus decreased after vaccination in rats exposed to ZEA. In conclusion, these results suggest that ZEA exposure via drinking water can cause an immunosuppressive effect by decreasing immunoglobulins in serum and cytokines in lymphoid organs.
Animals ; Antibody Formation ; Cytokines ; Drinking Water ; Granulocyte-Macrophage Colony-Stimulating Factor ; Immunoglobulin A ; Immunoglobulin E ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Interleukin-1 ; Interleukin-10 ; Interleukin-2 ; Interleukin-6 ; Lymph Nodes ; Parvovirus, Porcine ; Rats ; Spleen ; Thymus Gland ; Tumor Necrosis Factor-alpha ; Vaccination ; Zea mays ; Zearalenone

In order to obtain a virus-like particle vaccine both for porcine parvovirus (PPV) prevention and growth-promotion, VP2 gene of PPV NJ-a strain was amplified with PCR, and four copies of synthetic somatostatin gene were fused to the N-terminal of VP2 gene. The fused gene was cloned into pFast-HT A to construct the recombinant plasmid pFast-SS4-VP2, then the pFast-SS4-VP2 was transformed into DH10Bac competent cells and recombined with shuttle vector Bacmid, followed by identification with blue-white screening and PCR analysis for three cycles, and the positive recombinant was named as rBacmid-SS4-VP2. The positive Sf-9 cells were transfected with rBacmid-SS4-VP2 by Lipofectamine to produce recombinant baculovirus. When the cytopathic effect (CPE) was obvious, the transfected Sf-9 cell was harvested, and the positive recombinant virus was named as rBac-SS4-VP2. The insertion for the target gene into baculovirus genome was confirmed with PCR. SDS-PAGE and Western blotting revealed that the calculated protein of approximately 68 kDa was in the expressed in the insect cells. The Sf-9 cells infected with rBac-SS4-VP2 were stained positive against PPV antibody using the indirect immunofluorescence assay (IFA). Moreover, the virus particle self-assembly was observed under electron microscopy. 90 four-week-old mice were immunized by the recombinant protein coupled with different adjuvants alhydrogel, IMS and oil. VP2-specific ELISA antibodies, PPV-specific neutralizing antibody, somatostatin antibody and growth hormone levels were examined to evaluate the immunogenicity of this virus like particle. Results indicated that mice groups immunized rSS4-VP2 protein with alhydrogel and IMS developed similar humoral immune response comparing with inactived PPV vaccine. Mice group immunized with rSS4-VP2 generated higher level of SS antibody and growth hormone comparing with negative control, mice receiving rSS4-VP2 with alhydrogel developed the highest antibody titre than all other groups, while the oil group developed the lowest antibody level. This study provides not only a new rout for production of safe and effective virus like particle subunit vaccine, but also the foundations for peptide presentation and multivalent subunit vaccine design.
Animals ; Antigens, Viral ; biosynthesis ; genetics ; Artificial Gene Fusion ; Baculoviridae ; genetics ; Capsid Proteins ; biosynthesis ; genetics ; Mice ; Parvoviridae Infections ; prevention & control ; Parvovirus, Porcine ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Somatostatin ; genetics ; Swine ; Vaccines, Virus-Like Particle ; biosynthesis ; immunology ; Viral Vaccines ; biosynthesis ; immunology ; Virion ; genetics ; immunology

To construct secretory expression vector of PPV NS1 gene, the fragment of PPV NS1 gene coding for major antigen region of the NS1 protein was amplified by PCR and inserted into multiple clone site of eukaryotic expression vector pPICZalpha-A. The recombinant pPICZalpha-A-NS1 plasmid was transferred into P. pastoris strain GS115 mediated by electro transform. Recombinant P. pastoris strain GS115 was induced to express the fusion protein by methanol. The expressed and purified protein was analyzed by SDS-PAGE and Western Blot. The recombinant protein was highly-expressed and showed a good immunoreactivity. The indirect ELISA method was developed for detecting antibodies against PPV by checkerboard titration assay. The result showed that the optimal concentration of coated antigen was 3.2 microg/mL and the best dilution of serum was 1 : 80. The positive cut-off value of the ELISA assay was OD450 > 0.4 and OD450 positive serum/OD450 negative serum > 2.0. Compared with HI and commercial ELISA kits, the assay revealed 94.2% and 92.1% agreement respectively. The assay demonstrates good specificity and sensitivity, and can be applied in the detection of porcine parvovirus.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Parvoviridae Infections ; diagnosis ; immunology ; veterinary ; virology ; Parvovirus, Porcine ; genetics ; immunology ; isolation & purification ; Recombinant Proteins ; genetics ; immunology ; Swine ; Swine Diseases ; diagnosis ; immunology ; virology ; Viral Nonstructural Proteins ; genetics ; immunology

To construct gene vaccine of PPV and to investigate the effects of interleukin 2 (IL-2) as an adjuvant on immune responses in mouse, the recombinant expression plasmid of pCIneo-IL2-VP2 was constructed and transfected into PK-15 cells by lipofectamine, the expressed product was detected by immunofluore assay. To study the immune effects of DNA vaccine in vitro and in vivo, mice were used as the animal model. The recombinant plasmid pCIneo-IL2-VP2, the control plasmid pCI-neo and the PPV live vaccine were immunized by intramuscular injection. Anti-PPV antibodies were measured by ELISA, lymphocyte proliferation activity was detected using MTT method, and the specific killing activities of CTL were assayed too. The results show that the immunized mice produced PPV antibody after one week, and reached to highest after four weeks. Compared with the control group, the pCIneo-IL2-VP2 immunized group produced significant differences in the antibody titers, the lymphocyte proliferation activity and the specific killing activities of CTL. The pCIneo-IL2-VP2 induced humoral and cellular immunity responses similarly to that the live vaccine induced. These results manifested that the PPV DNA vaccine successfully induced humoral and cellular immunity response in mice with the IL-2 gene as an adjuvant.
Adjuvants, Immunologic ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Capsid Proteins ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Immunization ; Interleukin-2 ; genetics ; immunology ; Mice ; Parvovirus, Porcine ; genetics ; immunology ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology