Porcine Circovirus Diseases Research Articles

Natural recombinants derived from different patterns of recombination between two PCV2b parental strains

Porcine circovirus diseases (PCVD) are globally emerging diseases that have huge economic impacts on swine industry. Porcine circovirus type 2 (PCV2) is considered to be the essential primary causative agent of PCVD. In the present study, recombination analyses of PCV2 identified two possible recombination events with high confidence using recombination detection program, phylogenetic analysis and base-by-base comparison. These recombination events occurred between strains 09CQ (HQ395024) and ZhuJi2003 ( AY579893), giving rise to two recombinants 09GS (HQ395028) and HN0907 (GU938303). Phylogenetic analyses of the parental strains at full length level suggest that natural recombination happened between PCV2b strains. Interestingly, recombination of the two parental strains yielded two recombinants through different recombination patterns with crossover regions mainly located in ORF1 and ORF2, respectively. These results demonstrate that recombination between PCV2b strains can occur both in non-structural protein coding region and structural protein coding region. Our study not only indicates that PCV2b strains can undergo recombination through a variety of patterns, but also suggests that recombination contributes to the genetic diversity of PCV2.

Development of an in situ assay for simultaneous detection of the genomic and replicative form of PCV2 using padlock probes and rolling circle amplification

BackgroundIn this study we utilized padlock probes and rolling circle amplification as a mean to detect and study the replication of porcine circovirus type 2 (PCV2) in cultured cells and in infected tissue. Porcine circovirus type 2 is a single-stranded circular DNA virus associated with several severe diseases, porcine circovirus diseases (PCVD) in pigs, such as postweaning multisystemic wasting syndrome. The exact reason and mechanisms behind the trigger of PCV2 replication that is associated with these diseases is not well-known. The virus replicates with rolling circle replication and thus also exists as a double-stranded replicative form.ResultsBy applying padlock probes and rolling circle amplification we could not only visualise the viral genome but also discriminate between the genomic and the replicative strand in situ. The genomic strand existed in higher numbers than the replicative strand. The virus accumulated in certain nuclei but also spread into the cytoplasm of cells in the surrounding tissue. In cultured cells the average number of signals increased with time after infection.ConclusionsWe have developed a method for detection of both strands of PCV2 in situ that can be useful for studies of replication and in situ detection of PCV2 as well as of DNA viruses in general.

Distribution of ORF2 and ORF3 genotypes of porcine circovirus type 2 (PCV-2) in wild boars and domestic pigs in Germany

Porcine circovirus 2 (PCV-2), the essential infectious agent in PCVD (porcine circovirus diseases) circulates at high rates among domestic pig and wild boar populations. Wild boars may be viremic and shed the virus with excretions and secretions, and thus serve as a reservoir for domestic pig PCV-2 infection. We hypothesize that PCV-2 strains circulating in wild boars and in domestic pigs are significantly different and thus, partially independent. To prove this hypothesis, the present study investigated by sequence analysis the distribution of ORF2 and ORF3 genotypes of the PCV-2 genome within wild boars ( n = 40) and domestic pigs ( n = 60) from overlapping greater areas of Germany. The genotypes were compared with PCV-2 sequences from the Genbank database. The dominating genotype in domestic pigs was PCV-2b (98.4% of infected pigs), while only 4.8% of them were infected with PCV-2a. The corresponding prevalences of PCV-2a and -2b genotypes in wild boars were 58% and 70%, respectively. When also ORF3 genotypes were taken into account, more than 50% of wild boar PCV-2 genotypes were rare among German and European domestic pigs. In conclusion, these data provide evidence for a certain independence of PCV-2 infections in both species and a low chance for domestic pigs to be infected with PCV-2 of wild boar origin. On the other hand, PCV-2 genotypes specific for domestic pigs are also common in wild boars, although at lower frequencies, suggesting the spread of domestic pig PCV-2 to the wild boar population.

Genetic characterization and phylogenetic analysis of porcine circovirus type 2 strains present in Cuban swine herds

Porcine circovirus type 2 (PCV2) is the essential infectious agent of postweaning multisystemic wasting syndrome (PMWS) considered as one of the most important swine diseases worldwide. One of the main risk factors reported for the development of PMWS is the PCV2 genotype. The presence of PCV2 in Cuban swine herds has been reported recently. However, genetic information about these viruses is not available yet. Hence, the objectives of this study were to classify the Cuban porcine circovirus type 2 sequences as well as to investigate the genetic diversity and the putative origins of the virus circulating in Cuban swine herds. PCV2 Cuban sequences appeared to be close related when an analysis of the entire viral genome sequences was performed. The main variations on amino acid sequences of the capsid protein were found within the immunoreactive areas. All the Cuban PCV2 sequences analyzed belonged to genotype 1 and were located within the same Cluster (1A). Interestingly, five of them were clustered with high confident values with those described as the PCV2 variants associated with severe porcine circovirus diseases reported in Canada from the late 2004 to 2006. Pigs imported from this source appeared to be the most probable origin of the viruses circulating in Cuban swine herds currently. The fact that one sequence was not clustered with any other group of PCV2 within genotype 1 might suggest that different introductions of the agent in the country from unknown sources have occurred.

Occurrence and Tissue Distribution of Porcine Circovirus Type 2 Identified by Immunohistochemistry in Danish Finishing Pigs at Slaughter

Infection with porcine circovirus type 2 (PCV2) may be subclinical or lead to the development of porcine circovirus disease (PCVD), which includes the entities of post-weaning multisystemic wasting syndrome (PMWS) and the porcine respiratory disease complex (PRDC). PCV2 infection and PMWS occur in the early post-weaning period and are also recognized in finishing pigs of 12-19 weeks of age. The aim of the present study was to assess the role of PCV2 infection in disease of finishing pigs. Accordingly, the occurrence and tissue distribution of PCV2 was examined in Danish finishing pigs at the time of slaughter. Multiple lymph nodes and the spleen, lungs and kidneys from 136 pigs with PRDC (case group) and 36 pigs without lung lesions (control group) were examined by immunolabelling for the presence of PCV2. Additionally, follicular dendritic cells (FDC) were identified immunohistochemically. One or more tissues of 61% of the pigs were positive for PCV2 antigen. Up to 78% of the pigs had mild lymphoid depletion, indistinct lymphoid follicles and/or histiocytic infiltration of the lymph nodes, but these lesions were not associated with PCV2. No association was found between the presence of lung or kidney lesions and detection of PCV2. Three distinct patterns of cellular PCV2 antigen labelling were recognized: (1) labelling of cells with stellate morphology and reticular distribution, (2) labelling of isolated non-epithelial, cells, and (3) epithelial labelling. The reticular pattern was most common and localized to the centres of lymphoid follicles, corresponding to the morphology and distribution of FDCs. This observation may be interpreted to suggest that PCV2 may interact with FDCs to cause depletion of B lymphocytes. Alternatively, the FDCs may be a reservoir of infective PCV2 in subclinically infected animals or represent a simple storage site for PCV2 antigen in pigs that have recovered from infection.

Porcine circovirus 2 (PCV2) induces a procoagulant state in naturally infected swine and in cultured endothelial cells

Porcine circovirus 2 (PCV2) is the primary causative agent of porcine circovirus disease (PCVD). PCVD is an emerging disease that has been reported worldwide, associated with wasting, lymphoid depletion, enteritis, pneumonia, vasculitis, ischemic lesions, and necrotizing dermatitis. Although PCVD causes considerable economic losses, the pathogenesis of PCV2 has not been fully understood. The aim of the present work was to study the participation of hemostatic system and of vascular endothelium in PCV2 infection, as well as their possible role in PCVD pathogenesis. Our results showed that naturally PCV2-infected swine displayed a prothrombotic state in vivo, since a diminished coagulation time (recalcification time, activated partial thromboplastin time and prothrombin time), a higher platelet aggregation ability (despite a diminished platelet blood count), and an increased thrombin plasma activity (associated with a reduced fibrinogen level) were observed. The PCV2-infected animals showed vasculitis and positive staining for PCV2 antigen in capillary vessels. Furthermore, PCV2-infected endothelial cells displayed an activated phenotype, characterized by an increase in cell surface procoagulant activity. Moreover, the PCV2-infected endothelial cells pre-treated with exogenous thrombin displayed an increased viral load. This work reports, for the first time, the role of the hemostatic system and of endothelium in the pathogenesis and infectivity of PCV2. The study reinforces the importance of the phenomena which occur during PCV2 infection, and affords a better knowledge of the mechanisms behind the pathophysiology of PCVD.

Swine torque teno virus detection in pig commercial vaccines, enzymes for laboratory use and human drugs containing components of porcine origin

Torque teno viruses (TTVs) are vertebrate infecting, single-stranded circular DNA viruses. Two genetically distinct TTV genogroups (TTV1 and TTV2) infect swine worldwide with high prevalence. Currently, swine TTVs are considered non-pathogenic, although TTV2 has been linked to post-weaning multisystemic wasting syndrome, a porcine circovirus disease. On the other hand, pig materials are an important source of components used in porcine vaccine manufacturing, human drugs and commercial enzyme products. However, there is little information about the possible existence of extraneous viruses in products containing porcine-derived components. In the present study, 26 commercial swine vaccines, seven human drugs and three enzyme products from porcine origin were tested for the presence of TTV1 and TTV2 genomes by PCR. Four vaccines against Mycoplasma hyopneumoniae were positive for TTV2 by PCR. Three M. hyopneumoniae, one porcine parvovirus and one porcine reproductive and respiratory syndrome virus vaccines were PCR positive for TTV1. One human drug contained TTV1 DNA as well as a trypsin enzyme; a porcine-derived elastase product was positive for both TTV genogroups. These results show that swine TTVs are contaminants not only of swine vaccines but also of human drugs containing porcine components and enzymes for laboratory use.

A field evaluation of mortality rate and growth performance in pigs vaccinated against porcine circovirus type 2

To evaluate, under field conditions, the effects of a commercial porcine circovirus type 2 (PCV2) vaccine on mortality rate and growth performance in a herd infected with PCV2 that had a history of porcine circovirus disease. Randomized controlled clinical trial. 485 commercial, cross-bred, growing pigs. Prior to weaning, pigs were randomly assigned within litter to a vaccination or unvaccinated control group. Pigs in the vaccination group were given a commercial PCV2 vaccine at weaning and 3 weeks later. Mortality rate was recorded, and pigs were weighed prior to vaccination, when moved from the nursery, and prior to marketing. Infection status was assessed by serologic testing and detection of viral DNA in serum. Compared with control pigs, pigs vaccinated against PCV2 had a significantly lower mortality rate during the finishing phase, significantly higher average daily gain during the finishing phase, and significantly lower likelihood of being lightweight at the time of marketing. For vaccinated pigs, overall mortality rate was reduced by 50% and average daily gain during the finishing period was increased by 9.3%. At the time of marketing, vaccinated pigs weighed an average of 8.8 kg (19.4 lb) more than control pigs, without any difference in days to marketing. Serum PCV2 antibody titers increased in control pigs, and PCV2 DNA was detected, indicating active PCV2 infection. Results suggested that vaccination against PCV2 was effective at reducing mortality rate and improving growth performance among pigs in a herd infected with PCV2.

Evidence for recombination between PCV2a and PCV2b in the field

Genomic sequence analysis demonstrates that porcine circovirus type 2 (PCV2) isolates are divided into distinct genotypes. Historically, swine herds in the U.S. have been infected with the PCV2a genotype. In 2005, PCV2b was identified in North America and with it increased reports of porcine circovirus disease (PCVD). A differential PCR technique incorporating PCV2 genotype-specific primers was used in the clinical diagnosis of PCVD. A set of 97 diagnostic submissions showed that both PCV2a and PCV2b were present in 25% of clinical samples. The construction of phylogenetic trees using whole genome sequences from diagnostic submissions showed that one isolate, 0737A, was only loosely associated with other PCV2b isolates. Analysis of the variable sites between representative PCV2a and PCV2b DNA sequences and the 0737A sequence, showed that 0737A was a mosaic sequence, with the ORF1 region from PCV2a and ORF2 from PCV2b. This study demonstrates that pigs can be naturally infected with multiple PCV2 genotypes and that PCV2a/PCV2b recombination events occur in the field.

Genetic diversity of Brazilian strains of porcine circovirus type 2 (PCV-2) revealed by analysis of the cap gene (ORF-2)

Porcine circovirus 2 (PCV-2) is associated with a broad range of syndromes. In this study, 19 of 870 samples from pigs from different Brazilian states were found to be positive for PCV-2 by polymerase chain reaction (PCR). A fragment of 700 nt of the cap gene (ORF-2) from the 19 PCV-2-positive samples were sequenced using three pairs of primers (Fa/Ra, Fb/Rb and Fc/Rc). Maximum parsimony genealogy with a heuristic algorithm using the 19 field strain studied here, 21 sequences from GenBank and PCV-1 as an out-group showed the existence of two major clusters (A and B) and the Brazilian strains segregating in both of them. PCV-2 was found in pigs with various clinical signs. No association between clusters of PCV-2 and different states or clinical signs were observed, demonstrating that the exact role of PCV-2 in porcine circovirus diseases (PCVD) in Brazil still needs to be clarified. These results contribute to the molecular characterization of PCV-2, which serve as a basis for the epimiology of PCV-2 infection.

Effects of pcv2 vaccine on the growth performance of pigs and mortality rate in a pcv2-positive commercial swine herd (2007)

A total of 1,470 pigs were used to study a commercial sow herd with a history of Porcine Circovirus Disease (PCVD). The objective was to evaluate the effect of two commercially available Porcine Circovirus Type 2 (PCV2) vaccines on growth and mortality rates. The first vaccine was administered one week after weaning (1-dose) while the second was administered at weaning and repeated three weeks later (2-dose). A third group of unvaccinated pigs served as a control group. Pigs were individually weighed at weaning (d 0), d 113, 143, and just prior to market. On d 113, pigs on the 2-dose treatment were heavier (P<0.05) than the control group, and the 1dose treatment pigs were intermediate. At d 143, just prior to when the first pigs were marketed, both the 1-dose and the 2-dose pigs were heavier than the control pigs by 7.6 and 10.2 lb (P<0.05), respectively, and there were no significant differences in weights between the two vaccinated groups. However, differences in weights between the vaccinated and the control pigs were smaller at off-test compared to differences at d 143 due to a wider variability in on-test days as a result of multiple marketing days prior to end of the trial. Although there were no significant differences between the two vaccinated groups, ADG was greater (P<0.05) in all vaccinated pigs compared to non-vaccinated control pigs from d 0 to d 113, d 143, and at off-test. From d 113 to 143 and until the day they were taken off test, there were no differences in ADG, regardless of treatment. This suggests that the increase in growth rate in vaccinated pigs occurred during the period d 0 to 113. Barrows consistently exhibited greater ADG and heavier weights (P<0.05) than gilts throughout the trial. No significant differences in mortality rate between treatments were observed but both vaccinated groups had mortality rates that were 3% lower than the non-vaccinated control pigs. Based on these results, both commercial vaccines were effective in mitigating the effects of PCV2 virus and improving the growth performance of pigs in a PCV2 positive herd.

Estimation of European wild boar relative abundance and aggregation: a novel method in epidemiological risk assessment

Wild boars are important disease reservoirs. It is well known that abundance estimates are needed in wildlife epidemiology, but the expense and effort required to obtain them is prohibitive. We evaluated a simple method based on the frequency of faecal droppings found on transects (FBII), and developed a spatial aggregation index, based on the runs test statistic. Estimates were compared with hunting data, and with porcine circovirus and Aujeszky's disease virus seroprevalences and Mycobacterium tuberculosis complex and Metastrongylus spp. prevalence. The FBII and the aggregation index were correlated with the hunting index, but both of the former estimates correlated better than the latter with the disease prevalences. Hence, at least in habitats with high wild boar densities, the FBII combined with the aggregation index constitutes a cheap and reliable alternative for wild boar abundance estimation that can be used for epidemiological risk assessment, even outside the hunting season and in areas with no available data on hunting activities.

Porcine circovirus diseases

Porcine circovirus type 2 (PCV2) is a member of the family Circoviridae, a recently established virus family composed of small, non-enveloped viruses, with a circular, single-stranded DNA genome. PCV2, which is found all over the world in the domestic pig and probably the wild boar, has been recently associated with a number of disease syndromes, which have been collectively named porcine circovirus diseases (PCVD). Postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephropathy syndrome (PDNS) and reproductive disorders are the most relevant ones. Among them, only PMWS is considered to have a severe impact on domestic swine production. PMWS mainly affects nursery and/or fattening pigs; wasting is considered the most representative clinical sign in this disease. Diagnosis of this disease is confirmed by histopathological examination of lymphoid tissues and detection of a moderate to high amount of PCV2 in damaged tissues. Since PMWS is considered a multifactorial disease in which other factors in addition to PCV2 are needed in most cases to trigger the clinical disease, effective control measures have focused on the understanding of the co-factors involved in individual farms and the control or elimination of these triggers. PDNS, an immuno-complex disease characterized by fibrino-necrotizing glomerulonephritis and systemic necrotizing vasculitis, has been linked to PCV2, but a definitive proof of this association is still lacking. PCV2-associated reproductive disease seems to occur very sporadically under field conditions, but it has been characterized by late-term abortions and stillbirths, extensive fibrosing and/or necrotizing myocarditis in fetuses and the presence of moderate to high amounts of PCV2 in these lesions. Taking into account that scientific information on PCV2 and its associated diseases has been markedly expanded in the last 8 years, the objective of this review is to summarize the current state of knowledge of the most relevant aspects of PCV2 biology and PCVD.

A review of porcine circovirus 2-associated syndromes and diseases

Clinical expression of porcine circovirus 2 (PCV2) infection in swine may result in several distinct syndromes and diseases including post-weaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephropathy syndrome (PDNS), reproductive failure, porcine respiratory disease complex, granulomatous enteritis, necrotizing lymphadenitis, and possibly exudative epidermitis. Association of PCV2 with congenital tremor in piglets is still controversial. The extent of the involvement of PCV2 in swine disease other than PMWS is currently poorly understood. This review concentrates on PCV-2-associated syndromes and diseases other than PMWS.