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Porcine circovirus associated diseases (PCVAD) caused by PCV2 are responsible for severe economic losses in the swine industry. The mechanism of PCV2 replication has not been fully elucidated yet. PCV2 may be successfully rescued by means of either an infectious DNA clone containing the full length of the viral genomic DNA, or from PCV2-infected clinical tissues in PK15 cell culture. However, viruses harvested by both methods have low titres. In this study, PCV2 was prepared with a higher titre from PK15 cells infected by recombinant baculoviruses containing 1PCV2 (one stem-loop structure) or 1.1PCV2 (two stem-loop structure) genomic DNA copy. In addition, infectious DNA clones containing two stem-loop structures in either plasmid or baculovirus backbones are capable of generating a higher virus titre than the DNA clones with only one copy of stem-loop structure.

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PCV2 and PRRSV are two important pathogens of domestic swine. There is considerable evidence that the infection is also present in wild boars. Meat juice provides an alternative to serum for antibody testing, and it has been used in testing for many important porcine infectious diseases. Samples of brachial muscle were collected from 142 wild boars shot in different regions of Poland during the 2006/2007 and 2007/2008 hunting seasons. Meat juice harvested from muscle samples was tested using an ELISA test specific for PCV2 and PRRSV antibodies. Additionally, IgG and IgM antibodies specific for PCV2 were detected in order to estimate the status of the PCV2 infection. Only one of the tested meat juice samples was positive for PRRSV (0.7%), and 68 out of 142 (47.9%) samples were positive for PCV2. Of the positive animals, 4 (2.8%) had an antibody profile suggesting active infection, 2 (1.4%) early active infection, and 62 (43.7%) late infection. Also, a lack of association between the age of the animals and the presence of antibodies related to the infection was noticed.

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Acta Veterinaria Hungarica
Authors:
Zhi Deng
,
An Yuan
,
Wei Luo
,
Nai Wang
,
Qian Gong
,
Xing Yu
, and
Li Xue

To investigate porcine circovirus type 2b (PCV2b) transmission by contact and vertical infection in Kunming mice (an outbred mouse stock deriving from Swiss albino mice with a high ratio of gene heterozygosis), four mice in cage 6 were inoculated with PCV2b and 25 mice without any treatment were placed into cages 1 to 5 (five mice in each cage). Seven days after being infected, the PCV2-binoculated mice were co-mingled with non-inoculated mice from cages 1 to 5 successively at 7, 14, 21, 28 and 35 days post infection (dpi), respectively, for 3 days. In addition, eleven pregnant mice were injected with PCV2b. Samples were collected from non-inoculated mice and three newborn mice from each litter for PCV2b detection by polymerase chain reaction (PCR) and immunohistochemistry (IHC). The PCR results showed that PCV2b transmission rate among mice in cages 1, 2, 3, 4 and 5 was 0/5, 2/5, 5/5, 5/5 and 1/5, respectively. PCV2b antigen signals generally appeared in most organs of the non-inoculated mice in which viruses were detected by PCR. PCV2b DNA was also detected in newborn mice of PCV2b-infected litters, and viral antigen signals were observed in their organs as well. PCV2b was transmitted in Kunming mice by contact, and it also caused vertical infection through the placenta.

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Acta Veterinaria Hungarica
Authors:
Z. Lipej
,
J. Segalés
,
I. Toplak
,
B. Šoštarić
,
Besi Roić
,
M. Lojkić
,
P. Hostnik
,
J. Grom
,
Darja Barlič-Maganja
,
Kamelija Žarković
, and
D. Oraić

The objective of this study was to characterise porcine circovirus type 2 (PCV2) from pigs with naturally occurring postweaning multisystemic wasting syndrome (PMWS) in Croatia, and to determine the epizootiological, clinical and pathomorphological features of the disease. During a systematic health monitoring programme conducted in the period from January 2002 to June 2003, PMWS was suspected on eight different pig-producing farms in Croatia. The diagnosis of PMWS met all three key criteria: the presence of compatible clinical signs, the presence of the characteristic microscopic lymphoid lesions, and the detection of PCV2 within the lesions by polymerase chain reaction (PCR) and by in situ hybridisation (ISH). Moreover, PCV2 DNA from swine tissues was extracted and sequenced. The phylogenetic analysis of 4 Croatian PCV2 strains showed close relationship to PCV2 strains isolated in Slovenia, France, the Netherlands, the United Kingdom, China and Hungary. PCV2 was also demonstrated by electron microscopy in the lymph node of an affected animal. This is the first demonstration of PMWS in Croatia based on all scientifically accepted diagnostic criteria.

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This report describes the first case of postweaning multisystemic wasting syndrome (PMWS) in wild boar in Croatia. During the winter season of 2004, eight wild young piglets (of approximately 2 to 5 months of age) were found dead in a fenced hunting area. Polymerase chain reaction (PCR) was carried out on measenteric lymph nodes and all animals yielded positive results. In one of these animals diagnosis of PMWS was established based on the three key diagnostic criteria including the clinical manifestation, moderate lymphoid lesions consisting of lymphocyte depletion and granulomatous inflammation, and detection of the presence of PCV2 genome within the lymphoid lesions by in situ hybridisation (ISH). Three additional wild piglets had also mild PMWS-like lesions and a low amount of PCV2 was also found. No PMWS-like lesions or PCV2 genome were detected in the rest of the wild piglets studied. Three PCR-positive isolates were partially sequenced, which confirmed the diagnosis of PCV2 and demonstrated that the three sequences were genetically identical. The phylogenetic analysis of a representative PCV2 isolate indicated that its sequence (DQ875444) is grouped in a separate branch with Hungarian isolate (AY256460) and differs from any of the annotated sequences.

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Acta Veterinaria Hungarica
Authors:
Márta Lőrincz
,
Attila Cságola
,
Imre Biksi
,
Levente Szeredi
,
Ádám Dán
, and
Tamás Tuboly

Porcine circoviruses (PCV) are present worldwide, infecting domestic pigs and wild boars alike. Studies under laboratory conditions indicated that PCV can be taken up by mice and the virus can replicate in these animals. The possible role of rodents in maintaining and transmitting PCV2 infection in the field has not been investigated yet. The present study reports the detection of PCV2, the pathogenic form of the virus, in mice and rats. A number of rodents, such as mice, rats and voles, were collected at PCV2-infected farms and also outside pig herds and tested for the presence of the virus by polymerase chain reaction (PCR). The results indicated that PCV2 can be present both in mice and rats (65.0% and 23.8% positivity, respectively) on the infected premises, but those rodents that were collected outside pig farms remained negative for PCV2.

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Little information is known about infection, replication and transmission of porcine circovirus type 2 (PCV2) in species other than swine. Two sets of animal experiments were carried out to investigate the susceptibility of mice to PCV2 and to study their possible role in maintaining and transmitting the virus. In the first experiment 14 mice were inoculated with PCV2 by the intraperitoneal route with 5 × 10 2 TCID 50 of the PCV2-ROM strain (Cadar et al., 2007). In a second experiment 24 mice were divided into two groups (A and B); mice in Group A (n = 18) were inoculated orally with 1 × 10 5 TCID 50 PCV2-ROM and mice in Group B (n = 6) were left uninoculated until day 12 post inoculation (p.i.), when they were mixed with Group A. The animals were sacrificed at intervals for postmortem investigation and virus genome detection by polymerase chain reaction (PCR). The PCR results indicated that PCV2 could replicate in mice infected intraperitoneally or by the oral route, and that the virus can be transmitted directly from mouse to mouse.

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Acta Veterinaria Hungarica
Authors:
D. Cadar
,
A. Cságola
,
Á. Dán
,
Z. Deim
,
Marina Spînu
,
V. Miclăuş
,
L. Köbölkuti
,
G. Czirják
, and
T. Tuboly

Porcine circovirus type 2 (PCV2) has been demonstrated to be the causal agent for postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS). This report describes the first detection of PCV2 and associated diseases in a Romanian swine herd located in Transylvania. The clinical signs, pathological and histopathological changes observed in affected pigs were similar to those previously described for PDNS and PMWS. Polymerase chain reaction and in situ hybridisation were used for the detection of PCV2 nucleic acids from tissues and serum samples. Complete PCV2 genomes of both PMWS and PDNS cases were sequenced and analysed, and by comparing them with each other no genomic differences could be detected. The sequence analysis showed that the Romanian PCV2 was closely related to PCV2 identified in France and in Hungary.

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Acta Veterinaria Hungarica
Authors:
Daniel Cadar
,
Attila Cságola
,
Marina Spinu
,
Ádám Dán
,
Krisztina Ursu
,
Márta Lőrincz
, and
Tamás Tuboly

Porcine circoviruses (PCV) are widespread in domestic pigs worldwide and there is growing information about the presence of PCV in other suid species. Based on serological studies with sera of wild boars, it was established that PCV1 was present in these animals and antibodies specific to PCV2 were also detected in wild boars living in captivity or in sylvatic areas, both with or without clinical signs of PMWS. Studies including PCV2 genome or antigen detection confirmed the previous findings. This is the first report about the presence of PCV in Transylvanian wild boar populations. Four hundred and sixty-nine samples were collected and grouped according to geographic origin, tested for the presence of PCV DNA using a real-time quantitative polymerase chain reaction assay, and 13.52% of the animals proved to be positive for one or in three cases both of the PCV genotypes. PCV2 was detected in all of the PCV-positive samples.

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The aim of the present study was to explore the usefulness of serological methods in the diagnosis of postweaning multisystemic wasting syndrome (PMWS). The study was carried out in 4 PMWS-affected and 6 control farms. Based on the serological profiles, infection with porcine circovirus type 2 (PCV2) was determined to take place at 3–7 weeks of age in the PMWS-affected and at 3–11 weeks of age in the control farms. To compare the dynamics of seroconversion to PCV2 among farms, cross-sectional serological profiles were normalised in relation to the inferred age of infection. The results indicated that the proportion of seropositive pigs increased significantly slower in the affected herds. The most pronounced difference was observed about 4 weeks after infection, when the proportion of seropositive pigs ranged from 0 to 53.3% and from 70 to 100% in PMWS-affected and control herds, respectively. Mean antibody titres at that age were also significantly lower in the affected farms. These observations suggest a delay in the production of PCV2-specific antibodies and indicate that serological methods may be helpful in identifying herds with a high risk of PMWS.

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