Studies on porcine circovirus type 2 vaccination of 5-day-old piglets

Porcine circovirus type 2 (PCV2) vaccines have become widely used since they became available in 2006. It is not uncommon for producers to use PCV2 vaccines in pigs younger than what is approved by manufacturers. The objective of this study was to determine the efficacy of a chimeric and a subunit PCV2 vaccine administered at 5 or 21 days of age. Forty-eight PCV2-naïve piglets were randomly divided into six groups of eight pigs each. Vaccination was done at day 5 or day 21, followed by triple challenge with PCV2, porcine parvovirus (PPV), and porcine reproductive and respiratory syndrome virus (PRRSV) at day 49. Vaccinated pigs seroconverted to PCV2 approximately 14 days postvaccination and had a detectable neutralizing antibody response by 21 days postvaccination regardless of age at vaccination. At day 49, the pigs vaccinated with the chimeric vaccine had significantly higher levels of neutralizing antibodies than the pigs vaccinated with the subunit vaccine. After challenge, vaccinated pigs had significantly decreased levels of PCV2 viremia and a decreased prevalence and severity of microscopic lesions compared to the positive-control group, which had severe lymphoid lesions associated with abundant PCV2 antigen, compatible with PCV-associated disease. The results of this study indicate that, under the conditions of this study, vaccination of PCV2-naïve pigs at day 5 or day 21 resulted in development of a detectable humoral immune response and provided reduction or complete protection against PCV2 viremia and PCV2-associated lesions after triple challenge with PCV2, PPV, and PRRSV.     

Cell tropism and entry of porcine circovirus 2

Porcine circovirus 2 (PCV2) may induce reproductive failure (return to oestrus, embryonic death, mummification, weak- and stillborn piglets) and postweaning multisystemic wasting syndrome (PMWS). Furthermore, it may modulate the immunity in such a way that it aggravates the outcome of many bacterial and viral infections. In the present paper, the cellular tropism and entry of PCV2 are described and linked with the pathological and clinical consequences.     

Prevalence of porcine hokovirus and its co-infection with porcine circovirus 2 in China

Here, we report the frequency of porcine hokovirus (PHoV) infection and its co-infection with porcine circovirus 2 (PCV2) in China. A total of 485 domestic pig samples were tested for both PHoV and PCV2, and NS1 gene sequences from 11 PHoV strains were used for phylogenetic analysis. The prevalence of PHoV and PCV2 was 51.3 % and 36.3 %, respectively, and co-infection occurred in 20.2 %. PHoVs from the Chinese mainland showed a close relationship to those isolated in Hong Kong. Co-infection with both viruses was prevalent, and PHoV may contribute to the induction of postweaning multisystemic wasting syndrome (PMWS).     

Recombination of two porcine circovirus type 2 strains

Pigs can be concurrently infected with different PCV2 strains. In this study, a cell-culture-adapted PCV2 strain, originating from a PMWS-affected pig, was purified by limiting dilution. Three different strains were obtained, and one of them was a perfect mosaic of the other two, with recombination breakpoints in ORF1 and ORF2. Incongruence was observed between phylogenetic trees constructed with the whole genome, ORF1 and ORF2. Amplification of ORF1 and ORF2 from original material, followed by cloning and sequencing, resulted in sequences corresponding to the parental strains, but not with the mosaic strain. These results demonstrate that PCV2 can undergo recombination. The GenBank/EMBL/DDBJ accession numbers of the sequences from clones II11A (‘strain 1’), 4D4 (‘strain 3’) and II9F (‘strain 2’) determined in this study are EU909686, EU909687 and EU909688, respectively.     

Immunologic features of porcine circovirus type 2 infection

Clinical expression of porcine circovirus 2 (PCV-2) infection in swine may result in two distinct high mortality disease syndromes. In North America, postweaning multisystemic wasting syndrome (PMWS), while still sporadic in incidence, predominates. In Europe and elsewhere, both PMWS and a second syndrome, porcine dermatitis and nephropathy syndrome (PDNS), occur in endemic and epidemic forms. PMWS but not PDNS has been reproduced in piglets by inoculations with PCV-2 alone or in PCV-2-infected swine co-infected with porcine parvovirus (PPV) or porcine respiratory and reproductive syndrome (PRRS) virus and also if PCV-2-infected piglets are immunostimulated by injections with an immunogen emulsified in an oil-based macrophage-targeted adjuvant. Subclinical but active infection has been achieved by direct inoculation of piglets with cloned PCV-2 DNA and/or progeny virus derived from cloned DNA. Morphologic changes in lymphoid tissues and preliminary functional data suggest that immunosuppression may occur in PMWS-affected swine. This phenomenon appears to be mediated by generalized lymphoid depletion and replacement by infiltrating and proliferating histiocytes and macrophages. Accumulation of virus in both mononuclear phagocytes and follicular dendritic cells is characteristic of PCV-2 infection. Exogenous immunosuppression of PCV-2-infected gnotobiotic piglets with cyclosporine (Cys), but not corticosteroid (St), potentiates PCV-2 replication and promotes productive virus infection of hepatocytes in Cys-treated piglets, a tropism not previously apparent in experimentally induced PMWS in gnotobiotic swine. In the Cys-treated piglets, inflammatory lesions characteristic of PMWS are absent, even though tissues contain high titers of infectious virus, a finding which suggests that the granulomatous inflammatory lesions characteristic of PMWS are immune mediated.     

Studies on pathogenicity of turkey ureaplasma

Turkeys and chickens were experimentally infected with a ureaplasma strain T-1001 isolated from turkey semen. Following infection, sero-fibrinous airsacculitis and serological responses developed in turkeys and chickens of different ages.     

First analysis of synonymous codon usage in porcine circovirus

Porcine circovirus (PCV) is grouped into two types: PCV1 and PCV2. PCV1 is isolated from cultured cells and usually causes no clinical diseases in pigs. PCV2 is a pathogen of severe pig disease and a great threat to swine health and production. In our study, to investigate the codon usage bias of PCV, the genomic sequences of PCV1 and PCV2 were analyzed. The results showed that the codon usage bias of PCV was very low. An effective number of codons (ENC) plot analysis indicated that mutational pressure influences the codon usage bias of PCV. Neutrality plot analysis showed that mutation bias dominated over natural selection in shaping the codon usage bias of PCV1, but mutation bias and natural selection contributed equally to the codon usage bias of PCV2. Principal component analysis showed that different ORFs and dinucleotide patterns were also factors influencing the codon usage bias of PCV. Our study is helpful in understanding the codon usage pattern of PCV and the evolution of PCV.     

T lymphocyte epitope mapping of porcine circovirus type 2

Immunoreactive T lymphocyte epitopes within the ORF1, ORF2, and ORF 3 products of porcine circovirus type 2 (PCV2) were mapped. For this, overlapping linear 20-mer peptides were synthesized and tested for their ability to induce T lymphocyte proliferation in porcine peripheral blood mononuclear cells (PBMCs) isolated from experimentally PCV2-infected pigs. After a preliminary screening of 31 (ORF1), 23 (ORF2), and 10 (ORF3) peptides using PBMCs from 4 PCV2-infected pigs, none of the peptides appeared to be immunoreactive (stimulation index [SI] : 2) in all four pigs. Only 14 peptides appeared to be immunoreactive in 3 of the 4 pigs. These peptides were designated as immunodominant in the preliminary screening and selected for further analysis. The immunodominant peptides were resynthesized and purified by high-performance liquid chromatography and tested for their ability to induce T lymphocyte proliferation in PBMCs from another three PCV2-infected pigs. None of the immunodominant peptides appeared to be immunoreactive in all three pigs of the second screening. Only three peptides appeared to be immunoreactive in two of three pigs, two encoded by PCV2 ORF1 (amino acid residues 81-100 and 201-220) and one encoded by PCV2 ORF3 (amino acid residues 31-50), and were therefore considered to be immunodominant in both screenings. Although peptides encoded by ORF2 appeared to show the highest immunoreactivity in some pigs, none of these peptides displayed immunodominance in both screenings. In summary, the present study indicates that the T lymphocyte responses to PCV2 are primarily directed toward epitopes of the nonstructural proteins of ORF1 and ORF3.     

Distinction between porcine circovirus type 2 enteritis and porcine proliferative enteropathy caused by Lawsonia intracellularis

The presence of porcine circovirus type 2 (PCV2) was studied immunohistochemically in formalin-fixed, paraffin wax-embedded samples of intestinal tissue from 80 pigs with a clinical history suggestive of Lawsonia intracellularis-associated diarrhoea. Histopathologically, enteritis of varying intensity was diagnosed in 64 of the pigs. Of these 64 animals, 34 (18%) were infected with both PCV2 and L. intracellularis. Of the remaining 30 cases of enteritis, 23 (77%) were attributed to PCV2 infection alone. The PCV2-associated enteritis cases showed necrotizing ileitis and colitis, indistinguishable macroscopically from proliferative enteritis (PE) due to L. intracellularis. Histopathologically, L. intracellularis-positive intestines showed adenomatous proliferation of crypt enterocytes, whereas PCV2 enteritis was characterized by histiocytosis of varying intensity, with PCV2-positive cells in the submucosa, lamina propria and crypt epithelium, as well as in the lymphoid tissue of the ileum and colon. Multinucleated giant cells, however, were seen in both infections. PCV2 was about three times more likely to be detected in L. intracellularis-negative than in L. intracellularis-positive samples (P<0.001). There was no association between PCV2 and other intestinal bacterial pathogens. The study demonstrated that PCV2 enteritis should be borne in mind in the differential diagnosis of L. intracellularis infection in pigs aged 2-4 months with a clinical history of diarrhoea.     

Current understanding of genomic DNA of porcine circovirus type 2

Porcine circovirus type 2 (PCV2) is the primary causative agent of porcine circovirus-associated diseases in swine and is also described as the modulator of host immunity that exacerbates the clinical outcome of many bacterial and viral infections. To date, it has caused increasingly larger losses in the pig industry worldwide. The genomic DNA of PCV2 is predicted to contain 11 open reading frames (ORFs) and at least seven potential ORFs-encoding proteins larger than 5 kDa. Currently, however, only five virally encoded proteins (Rep, Rep′, Cap, ORF3, and ORF4 protein) have been identified in PCV2 replication. In the present review, we strive to discuss the current understanding of the genomic DNA of PCV2 with the purpose of providing insight into the scientific basis of the pathogenesis of PCV2 and the prevention of its infection.     

Molecular epidemiology of Korean porcine sapeloviruses

To evaluate the prevalence and genetic diversity of porcine sapeloviruses (PSVs) in Korea, a total of 100 diarrhea fecal samples from pigs were analyzed by RT-PCR and nested PCR assays with primer pairs specific for the VP1 gene. Overall, 34 % of the diarrhea samples tested positive for PSV, and a high proportion of infections occurred along with a variety of other enteric viruses and bacteria. Genomic and phylogenetic analysis of the VP1 genes revealed pronounced genetic diversities between PSVs from Korean and elsewhere. Our results indicate that PSV infections are very common in Korean pigs with diarrhea. The infecting strains are genetically diverse.     

Isolation and characterization of porcine circovirus 2 from cases of sow abortion and porcine dermatitis and nephropathy syndrome

Summary.  We report the isolation and characterisation of porcine circovirus 2 (PCV2) from cases of sow abortion and porcine dermatitis and nephropathy syndrome. The results suggest that the clinical scope of PCV2 infections requires continuous re-evaluation. Received September 13, 2000 Accepted December 19, 2000