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Porcine circovirus 2 (PCV2) is a major etiological agent for porcine circovirus-associated diseases and causes enormous economic losses in domestic and overseas swine production. However, there are currently no suitable cell models to study the cytopathic effects (CPE) of PCV2 in vitro, which severely restricts the study of PCV2 pathogenesis. In the present study, we established an immortalized porcine oral mucosal epithelial cell line (hTERT-POMEC) by introducing the hTERT gene into primary porcine oral mucosal epithelial cells (POMECs) derived from a neonatal, unsuckled piglet. The hTERT-POMEC cells have a homogeneous cobblestone-like morphology and retain the basic physiological properties of primary POMECs. No chromosome abnormality and tumorigenicity transformation was observed in immortalized hTERT-POMECs. Viral infection assays demonstrated that PCV2 propagated and caused CPE in hTERT-POMECs. We conclude that the immortalized cell line hTERT-POMEC is a crucial tool for further research into the pathogenesis of PCV2.

The presence of porcine circovirus 2 and porcine parvovirus was examined in forty clinical samples of spleen, lymph nodes and lungs originating from non-vaccinated swine by polymerase chain reaction. All animals were reared in extensive livestock farming systems in different geographical districts of Republic of Srpska, Bosnia and Herzegovina. Porcine circovirus 2 DNA was detected in four lymph node and two spleen samples (15%), while porcine parvovirus DNA was identified in five lymph node samples (12.5%). The presence of both viruses was detected in three lymph node samples (7.5%). Partial nucleotide sequence of ORF1 gene of 2 porcine circovirus 2 and VP2 gene of 2 porcine parvovirus isolates was determined. The nucleotide sequences of two PCV2 isolates from RS-BIH included in phylogenetic typing are similar and cluster together with the strain Mantova isolated from domestic pigs in Italy, strains DE006-14 and DE222-13 isolated from pigs in Germany as well as with the strain Jvnan isolated from pigs in China. Also, analyzed PCV2 isolates were partially similar to the strain NIV-C SRB isolated from pigs in Serbia. The nucleotide sequences of two PPV isolates that were included in phylogenetic typing showed a high level of similarity with the strain Challenge isolated from pigs in UK, strain Kresse isolated from pigs in USA and strains 77 and LZ isolated from pigs in China. Prisustvo svinjskog cirkovirusa 2 i parvovirusa svinja ispitano je u četrdeset uzoraka (slezina, limfni čvorovi, pluća) poreklom od nevakcinisanih svinja primenom lančane reakcije polimeraze. Sve životinje su bile iz ekstenzivnog načina gajenja i iz različitih regiona Republike Srpske, BiH. Četiri uzorka limfnih čvorova i dva uzorka slezine su bili pozitivni na prisustvo DNK svinjskog cirkovirusa 2 (15%), dok je kod pet uzoraka limfnih čvorova utvrđeno prisustvo DNK parvovirusa svinja (12.5%). U uzorcima poreklom od tri svinje utvrđeno je prisustvo nukleinske kiseline oba prethodno navedena virusa (7.5%). Metodom sekvenciranja određena je nukleotidna sekvenca dela ORF1 gena dva izolata svinjskog cirkovirusa 2 i dela VP2 gena dva izolata parvovirusa svinja. Nukleotidne sekvence dva izolata PCV2 utvrđena u uzorcima svinja poreklom iz RS-BiH koja su bila uključena u filogenetsku analizu su pokazale visok stepen sličnosti sa nukleotidnim sekvencama soja Mantova izolovanog kod svinja u Italiji, zatim sojeva DE006-14 i DE222-13 izolovanih kod svinja u Nemačkoj kao i sa sojem Jvnan izolovanog kod svinja u Kini. Istovremeno, izolati PCV2 utvrđeni kod svinja u RS-BiH su bili delimično slični sa sojem NIV-C SRB virusa PCV2 izolovanim kod svinja u Srbiji. Nukleotidne sekvence dva izolata parvovirusa svinja uključenih u filogenetsku analizu su pokazale visok stepen sličnosti sa sojem Challenge izolovanim kod svinja u UK, sojem Kresse izolovanim kod svinja u SAD-u kao i sojevima 77 i LZ izolovanim kod svinja u Kini.

Prisustvo svinjskog cirkovirusa 2 i parvovirusa svinja ispitano je u osamdeset zbirnih uzoraka (slezina, limfni čvorovi, pluća) poreklom od nevakcinisanih svinja primenom lančane reakcije polimeraze. Sve životinje su bile iz ekstenzivnog načina gajenja i iz različitih regiona Republike Srpske, BiH. Četiri uzorka limfnih čvorova i dva uzorka slezine su bili pozitivni na prisustvo DNK svinjskog cirkovirusa 2 (7,5%), dok je kod pet uzoraka limfnih čvorova utvrđeno prisustvo DNK parvovirusa svinja (6,25%). U uzorcima poreklom od tri svinje utvrđeno je prisustvo nukleinske kiseline oba prethodno navedena virusa (3,75%). Metodom sekvenciranja određena je nukleotidna sekvenca dela ORF1 regiona genoma svih izolata svinjskog cirkovirusa 2 i dela VP2 gena identifikovanih sojeva parvovirusa svinja. Nukleotidne sekvence virusa PCV2 utvrđene u uzorcima svinja poreklom iz RSBiH uključene u filogenetsku analizu su pokazale visok stepen sličnosti sa nukleotidnim sekvencama soja Mantova izolovanog kod svinja u Italiji, zatim sojeva DE00614 i DE22213 izolovanih kod svinja u Nemačkoj kao i sa sojem Jvnan izolovanog kod svinja u Kini. Istovremeno, virusi PCV2 utvrđeni kod svinja u RSBiH su bili delimično slični sa sojem NIVC SRB virusa PCV2 izolovanim kod svinja u Srbiji. Nukleotidnesekvence svinjskih parvovirusa identifikovanih kod svinja u Republici Srpskoj uključene u filogenetsku analizu su pokazale visok stepen sličnosti sa sojem analognim sekvencama sojeva Challenge izolovanim kod svinja u UK, Kresse izolovanim kod svinja u SADu kao i sa sojevima 77 i LZ izolovanim kod svinja u Kini... The presence of porcine circovirus 2 and porcine parvovirus was examined in eighty clinical samples of spleen, lymph nodes and lungs originating from nonvaccinated swine by polymerase chain reaction. All animals were reared in extensive livestock farming systems in different geographical districts of Republic of Srpska, Bosnia and Herzegovina. Porcine circovirus 2 DNA was detected in four lymph node and two spleen samples (7.5%), while porcine parvovirus DNA was identified in five lymph node samples (6.25%). The presence of both viruses was detected in three lymph node samples (3.75%). Partial nucleotide sequence of ORF1 gene of porcine circovirus 2 and VP2 gene of porcine parvovirus isolates was determined. The nucleotide sequences of identified PCV2 viruses from RSBIH included in phylogenetic typing are similar and cluster together with strain Mantova isolated from domestic pigs in Italy, strains DE00614 and DE22213 isolated from pigs in Germany as well as with the strain Jvnan isolated from pigs in China. Also, analyzed PCV2 isolates were partially similar with the strain NIVC SRB isolated from pigs in Serbia. The nucleotide sequences of identified PPV viruses that were included in phylogenetic typing showed a high level of similarity with the strain Challenge isolated from pigs in UK, strain Kresse isolated from pigs in USA and strains 77 and LZ isolated from pigs in China.

Porcine circovirus type 2 (PCV2) induces autophagy via the 5' adenosine monophosphate-activated protein kinase (AMPK)/extracellular signal-regulated kinase (ERK)/tuberous sclerosis complex 2 (TSC2)/mammalian target of rapamycin (mTOR) pathway in pig kidney PK-15 cells. However, the underlying mechanisms of AMPK activation in autophagy induction remain unknown. With specific inhibitors and RNA interference (RNAi), we show that PCV2 infection upregulated calcium/calmodulin-dependent protein kinase kinase-beta (CaMKKβ) by increasing cytosolic Ca(2+) via inositol 1,4,5-trisphosphate receptor (IP3R). Elevation of cytosolic calcium ion (Ca(2+)) did not seem to involve inositol 1,4,5-trisphosphate (IP3) release from phosphatidylinositol 4,5-bisphosphate (PIP2) by phosphoinositide phospholipase C-gamma (PLC-γ). CaMKKβ then activated both AMPK and calcium/calmodulin-dependent protein kinase I (CaMKI). PCV2 employed CaMKI and Trp-Asp (WD) repeat domain phosphoinositide-interacting protein 1 (WIPI1) as another pathway additional to AMPK signaling in autophagy initiation. Our findings could help better understanding of the signaling pathways of autophagy induction as part of PCV2 pathogenesis. Further research is warranted to study if PCV2 interacts directly with IP3R or indirectly with the molecules that antagonize IP3R activity responsible for increased cytosolic Ca(2+) both in PK-15 cells and PCV2-targeted primary cells from pigs.

The presence of pseudorabies virus (PrV), porcine parvovirus (PPV) and porcine circovirus 2 (PCV2) was examined in sixty samples (spleen and lymph nodes) and thirty samples of sacral ganglia collected from non-vaccinated swine by virus isolation and polymerase chain reaction (PCR). Using PCR method PrV was detected in three samples, PPV in seven samples and six samples were found positive for PCV2. The phylogenetic analysis of the nucleotide sequences of three PrV isolates identified in this study showed high similarity and significant clustering within the PrV genotype I strains such as Kaplan and Bartha isolated from pigs in Hungary, strain Becker isolated in USA and strain Kolchis isolated in Greece. The nucleotide sequences of two PPV isolates showed high level of similarity with the strain Challenge isolated from pigs in UK, strain Kresse isolated in USA and strains 77 and LZ isolated in China. The phylogenetic analysis of the nucleotide sequences of two PCV2 isolates showed high level of similarity and significant clustering within genotype PCV2b strains such as NIVS-3, NIVS-5 and NIVS-6 isolated in Serbia, strain 3959 isolated in Austria, strain PM165 isolated from pigs in Brasil, and strain XT2008 isolated in China. The results of our study present the molecular characterization of PrV, PPV and PCV2 identified in swine in Republic of Montenegro. Besides that, these results confirmed that PCR is a very useful method for rapid detection of these viruses in subclinically infected swine. Primenom metode izolacije virusa i PCR metode, ukupno je ispitano devedeset uzoraka poreklom od svinja iz ekstenzivnog uzgoja iz različitih delova Crne Gore na prisustvo pseudorabijes virusa svinja (PrV), parvovirusa svinja (PPV) i svinjskog cirkovirusa 2 (PCV2). Primenom PCR metode prisustvo PrV je ustanovljeno kod tri uzorka, PPV kod sedam uzoraka, dok je prisustvo PCV2 utvrđeno kod šest uzoraka poreklom od svinja. Nukleotidne sekvence tri izolata PrV utvrđene u uzorcima poreklom od svinja koje su bile uključene u filogenetsku analizu su pokazale visok stepen sličnosti sa nukleotidnim sekvencama sojeva Kaplan i Bartha virusa izolovanih u Mađarskoj, zatim sa sekvencom soja Becker izolovanog u SAD-u i sekvencom soja Kolchis izolovanog u uzorcima poreklom od svinja u Grčkoj. Sva tri izolata su pripadala genotipu I pseudorabijes virusa. Nukleotidne sekvence dva izolata parvovirusa svinja su pokazale visok stepen sličnosti sa sekvencom soja Challenge izolovanog kod svinja u Velikoj Britaniji, zatim sekvencama soja Kresse izolovanog kod svinja u SAD-u i sojeva 77 i LZ izolovanih kod svinja u Kini. Filogenetska analiza nukleotidnih sekvenci dva PCV2 izolata je pokazala visok stepen sličnosti sa sojevima genotipa PCV2b kao što su NIVS-3, NIVS-5 i NIVS-6 izolovanim u Srbiji, sojem 3959 virusa izolovanim u Austriji i sojem PM165 virusa izolovanim u Brazilu i sojem XT2008 virusa izolovanim u Kini. Dobijeni rezultati ispitivanja pružili su uvid u molekularnu karakterizaciju sojeva virusa Aujeckijeve bolesti, svinjskog parvovirusa i svinjskog cirkovirusa 2 identifikovanih kod svinja u Crnoj Gori. Pored toga, dobijeni rezultati ispitivanja su potvrdili opravdanost korišćenja metode PCR u brzoj i pouzdanoj detekciji prethodno navedenih virusa kod supklinički inficiranih svinja.

To the Editor: Cases of sporadic, locally acquired hepatitis E have been increasingly identified in industrialized countries over the last few years (1). In this setting, hepatitis E is thought to be a zoonotic infection, with pigs as the primary host. Consumption of uncooked or lightly cooked pork meat products is thought to be a key route of infection, but other routes of transmission have been documented (2). For example, there have been several iatrogenic cases after transfusion of hepatitis E virus (HEV)–contaminated blood products (3) and transplantation of an HEV-infected donor liver (4). However, in most cases the source and route of infection are uncertain. In May 2011, a 42-year-old woman sought care at the Royal Cornwall Hospital in Truro, United Kingdom, for a 1-week history of malaise, diarrhea, nausea, and vomiting. Physical examination results were normal. Her liver function test results, however, indicated hepatitis: alanine aminotransferase 2,785 IU/L (reference range 10–36 IU/L), alkaline phosphatase 319 IU/L (reference range 30–130 IU/L), and bilirubin 30 μmol/L (reference range

The aim of this study was to characterize the porcine circovirus 2 (PCV2) infections in farrowing sows and to evaluate an association with piglet viremia and weight. Twenty sows and 100 newborn piglets were studied. Colostrum and serum of the sows were obtained on the day of parturition. Milk samples were collected on day 20 postpartum. Blood samples were taken and the piglets were weighed on days 1, 20, 42, 63 and 84 postpartum. Colostrum, milk and serum were evaluated for PCV2 DNA load. Serum was evaluated for neutralizing antibodies. PCV2 DNA was found in 17/20 serum samples, 14/20 colostrum samples and 11/20 milk samples. On day 1 postpartum 29% of piglets were viremic. PCV2 viral load ranged from 3.02 to 6.75 log10 copies/mL considering all sampled days. There was no correlation between sow viremia, antibody levels or PCV2 load in colostrum and piglet viremia on day 1 postpartum. The PCV2 load in colostrum and milk was associated with viremia in piglets from weaning to 84 days postpartum. Piglets' PCV2 viremia and viral load could not be associated with weight throughout this study. O objetivo deste estudo foi caracterizar o efeito do infecção pelo circovírus suíno 2 (PCV2) em porcas gestantes na viremia e no peso da leitegada. Vinte porcas e 100 leitões recém-nascidos foram acompanhados. Amostras de colostro e soro das porcas foram obtidas no dia do parto. Amostras de leite foram coletadas no dia pós-parto 20. Os leitões foram pesados e tiveram amostras de soro coletadas nos dias um, 20, 42, 63 e 84 pós-parto. Soro, colostro e leite foram testados para carga viral do PCV2. Soro foi avaliado para presença de anticorpos neutralizantes. O DNA do PCV2 foi encontrado em 14 de 20 amostras de colostro e em 11 de 20 amostras de leite. No dia pós-parto 1, 29% dos leitões foram virêmicos. A carga viral do PCV2 variou 3,02-6,75 log10 cópias / mL, considerando todos os dias amostrados. Não houve correlação entre viremia das porcas e os níveis de anticorpos no soro ou na carga de PCV2 no colostro e na viremia dos leitões com um dia de vida. A carga de PCV2 no colostro e no leite foi associada à viremia em leitões do desmame até 84 dias pós-parto. A carga viral do PCV2 em leitões não foi associada com o peso ao longo deste estudo.

Fundação de Amparo a Pesquisa do Estado de Minas Gerais The diseases associated with PCV2 (PCVAD) require various factors to occur, however, the virus infection is critical to the development of any one of the syndromes. The present study aimed to determine differences in serologic and viremic profiles for PCV2 in the swine breed Piau and a commercial line (Landrace x Large White x Pietrain) at a subclinically infected farm by the virus studied. The experiment was conducted at the Genetic Improvement Pig Farm (GMS), Federal University of Viçosa (UFV), in which it isn´t carried out vaccination against PCV2. This study conducted a cross-sectional sample of sows (> 2 parity), pigs for 1-3 weeks, 3-8 weeks and 8-22 weeks of age. The serum samples were used to measure the level of total antibodies by ELISA and quantitation of viremia by real time PCR. The results showed that, at the age of 3-8 weeks, the Piau breed piglets seroconverted earlier than the commercial line piglets and the Piau breed sows showed lower levels of total antibodies in relation to the commercial line. There were no differences in viremia between the different stages of production within each genetic group or between groups. This work provides evidence that the breed Piau has a different humoral immune response than the commercial line studied when facing a natural PCV2 subclinical infection. The results of this study reinforce the importance of the conservation of native breeds that have not been used for development of high productivity commercial lines. As doenças associadas ao PCV2 (PCVAD) necessitam de vários fatores para ocorrer, no entanto, a infecção pelo vírus é fundamental para o desenvolvimento de qualquer uma das síndromes. O presente estudo teve como objetivo verificar diferenças nos perfis sorológicos e virêmicos para o PCV2 entre suínos da raça Piau e de uma linhagem comercial (Landrace x Large White x Pietrain) em uma granja subclinicamente infectada pelo vírus estudado. O experimento foi realizado na Granja de Melhoramento Genético (GMS) da Universidade Federal de Viçosa (UFV), na qual não é realizada a vacinação contra o PCV2. O presente estudo realizou uma amostragem transversal em porcas (>2º parto), suínos de 1-3 semanas, 3-8 semanas e 8-22 semanas de idade. As amostras de soro obtidas foram utilizadas para mensuração dos níveis de anticorpos totais por ELISA indireto e quantificação da viremia por PCR em tempo real. Os resultados demonstraram que, na idade de 3-8 semanas, os leitões da raça Piau soroconverteram mais precocemente em relação aos leitões da linhagem comercial e as porcas da raça Piau apresentaram menores níveis de anticorpos totais em relação às da linhagem comercial. Não houve diferença na viremia entre as diferentes fases de produção dentro de cada grupo genético ou entre os grupos. Este trabalho fornece indícios de que a raça Piau apresenta uma resposta imune humoral diferente da desenvolvida pela linhagem comercial estudada diante de uma infecção subclínica natural pelo PCV2. Os resultados obtidos neste estudo reforçam a importância da conservação das raças nativas que não foram utilizadas para formação de linhagens de alta produtividade.

Porcine circovirus associated disease (PCVAD) is currently one of the most economically important diseases facing the global swine industry. Porcine circovirus type 2 (PCV2) is the primary and essential causative agent of PCVAD, but development of clinical disease typically requires co-infection with other swine pathogens such as porcine reproductive and respiratory syndrome virus (PRRSV). The specific mechanisms of co-infection that lead to clinical disease are not fully understood, but immune modulation by the co-infecting viruses is thought to play a critical role. The ability of dendritic cells (DC) infected with PRRSV, PCV2 or both to induce regulatory T cells (Tregs) was evaluated in vitro. DCs infected with PCV2 significantly increased CD4+CD25+FoxP3+ Tregs (p

Porcine circovirus-2 (PCV2) has been related as the causative agent of the Postweaning Multissystemic Wasting Syndrome (PMWS) and other diseases called porcine circovirus associated diseases (PCVAD). They are responsible for economic losses in pork production worldwide. There is only a few scientific studies describing the infection in other species but swine and their hole at the epidemiologic dynamics of the diseases related to the PCV2. The aim of this study is to investigate the occurrence of infection by the PCV2 in wild mice (Mus musculus and Rattus rattus) captured in hog farms. The capture of the 40 sorted mice was made at 5 pig wean tofinish farms in Minas Gerais, an important state of pork production in Brazil. Samples of tissues (lymph nodes, spleen, kidney, heart and lungs) and blood were collected from the mice. The tissue fragments collected were submitted to immunohistochemistry and Nested PCR. Additionally, samples from spleen and lungs were analyzed by histology assays. Presence of antibodies anti-PCV2 was tested by ELISA assays. Immunohistochemical analysis showed positive prints in 12 animals, mostly on spleen (sub scapular area), lungs (alveolar macrophages) and kidney (inside the tubules). The 12 serum analyzed by ELISA hasn t detected antibodies anti-PCV2. Histopathological analyses revealed in some samples, a multifocal and lympho-neutrophilic interstitial bronchopneumonia, with some node formations. Moreover, spleen samples showed a mild to moderate lymphocyte depletion related to the PCVAD. The Nested PCR assays showed the presence of viral DNA at different tissues from 6 tested rodents. Thus, the results found in this work, indicate that mice from the species Mus musculus and Rattus rattus can be naturally infected by the PCV2 and they would play a hole in the epidemiology of PCVAD. However, more studies are necessary to confirm the transmission of the PCV2 from wild rodents to pigs. O porcine circovirus-2 (PCV2) é atribuído como um dos agentes relacionados a doenças associadas ao circovírus (PCVAD), ocasionando perdas econômicas significativas na produção mundial de suínos. Poucos trabalhos são realizados a respeito da infecção em outras espécies pelo PCV2 e sua participação na epidemiologia das doenças associadas ao vírus. O propósito desse estudo foi investigar a ocorrência de infecção em roedores peridomésticos das espécies Mus musculus e Rattus rattus pelo PCV2 em granjas comerciais de suínos. Animais dessas espécies foram capturados em importantes centros de produção no estado de Minas Gerais. Amostras de órgãos (linfonodos, baço, rins, fígado, pulmão) e sangue foram coletadas. Os fragmentos de tecidos coletados foram submetidos ao teste de imunohistoquímica e Nested PCR. Adicionalmente, foram realizadas avaliações histológicas em amostras de baço, rim e pulmão. Presença de anticorpos anti-PCV2 foram avaliados pela técnica de ELISA. O teste de imunohistoquímica demonstrou marcações encontradas em 12 animais, principalmente no baço (região subcapsular), no pulmão (macrófagos alveolares) e nos rins (interior dos túbulos). A análise do soro pela técnica de ELISA não detectou anticorpos contra o PCV-2 nas 12 amostras avaliadas.. A histopatologia demonstrou em algumas amostras, uma pneumonia bronco-intersticial neutrofílica e linfocítica, multifocal e moderada, com formação de nódulos linfóides associados a vasos e bronquíolos. No ensaio de nested-PCR foi detectado DNA viral em diferentes tecidos avaliados de seis animais. Os resultados citados demonstram que os roedores domésticos das espécies estudadas podem exercer importante papel na epidemiologia das doenças relacionadas ao PCV2. No entanto, mais estudos são necessários para comprovar a transmissão do PCV2 dos roedores para os suínos.

Fundação de Amparo a Pesquisa do Estado de Minas Gerais The Porcine circovirus 2 (PCV-2) virus is a small, non-enveloped and belonging to the Circoviridae family by presenting icosahedral symmetry and circular genome. The PCV-2 is the primary causative agent of the postweaning multisystemic wasting syndrome (PMWS), one of the most important emerging disease of pigs in the world, identified in 1991 in Specific Pathogen Free (SPF) pigs and first described in 1997. The PCV-2 is associated with several other syndromes, which led to a new name for the disease: "PCV-2-associated disease" (PCVAD). As important as the clinical disease, subclinical infection has gained great importance in studies of porcine. The objective of this study was to evaluate the productive impact of the subclinical infection in slaughter healthy animals. For this, were collected serum samples and inguinal lymph nodes individual of sixty-four slaughter pigs. The viral load in lymph and serum were quantified by real-time PCR and neutralizing antibodies (NA) were measured by virus neutralization test (VNT). The lymph nodes were examined for lymphoid depletion, histiocytes proliferation, the presence of multinucleated giant cells and evaluated without significant injury (SLS), stage I (mild injury), II (moderate injury) or III (severe injury). The viral load present in the inguinal lymph nodes and sera ranged from 103,82 to 106,77 PCV-2 copies/500ng total DNA and 0 to 102,89 PCV-2 copies/5μL DNA, respectively. There was no significant difference between the values of viral loads considering gender. There was also no significant correlation between viral loads and the productive analysis. Among the animals tested, 53.13% showed no significant lesions in the lymph node, 37.5% were classified as stage I and 9.37% in stage II lesion. The NA title was variable among the pigs tested. The titles ranged from 64 to 2048, the latter being represented by 65.62% of the animals. It was observed a significant difference between the values of viral load in lymph nodes in relation to evidence of NA (p

The porcine circovirus, caused by the porcine circuvirus type 2 (PCV 2), and the porcine enzootic pneumonia, caused by the Mycoplasma hyopneumoniae, are infectious contagious diseases of great importance for worldwide swine production, both causes pneumonias with different presentation patterns. They have cosmopolite distribution and are among the major causes in economic losses. This work was made aiming the evaluation of the co-infection between porcine circovirus 2 (PCV2) and Mycoplasma hyopneumoniae using real time polymerase chain reaction (qPCR), immunohistochemistry and histopathological analyses. 120 lungs fragments, 45 being from fragments with macroscopic lesions (CLM) and 75 being from fragments without macroscopic lesions (SLM) were analyzed by histopathology procedures. Among these, 32 samples were randomly selected (being 16 without macroscopic lesions and 16 with macroscopic lesions) for the immunohistochemistry analyses (for Mycoplasma hyopneumoniae detection) and qPCR (for PCV 2 detection). No significant difference was observed between the groups (SLM and CLM) on the tests, indicating that the absence of macroscopic lesions doesn t reject the possible presence of agents neither the presence of microscopic lesions. The histopatological technique showed itself as an important tool for diagnosis approach. Although the lesion are not patognomonic for the agents in the analyses, they are quite suggestive about the viral and bacterium scene, and these findings, together with the presence of clinical signs and agent detection (more frequently made by qPCR and immunohistochemistry), confirm the diagnosis of each agent isolated or co-infection. Although the samples analyzed by RT-PCR had shown higher number of negative results and low viral quantity, many authors related that lungs are among the organs with the lowest viral quantities of PCV2, being, for that reason, the liver linfonodes, the main target of the PCV2, more indicated for viral quantification. It was verified a positive correlation between the immunohistochemistry analysis and the histopathology analysis using the Pearson statistical test, which confirms the presence of the agent in the histopathology observations. Forward studies using linfonodes samples (target of the PCV2) and samples collected from different lung anatomic regions must be realized for evaluation of a co-infection for more accurate results. A circovirose suína, causada pelo circovirus suíno tipo 2 (PCV2), e a pneumonia enzoótica suína, causada pelo Mycoplasma hyopneumoniae, são doenças infecto-contagiosas de grande importância na suinocultura mundial, ambas causam pneumonias com diferentes formas de apresentação. Têm distribuição cosmopolita e estão entre as principais causas de perdas econômicas. O presente trabalho foi realizado com o objeto de avaliar a coinfecção entre Circovírus suíno 2 e o Mycoplasma hyopneumoniae, por meio da reação da polimerase em cadeia em tempo real (qPCR), técnica de imunoistoquímica e exames histopatológicos. Cento e vinte fragmentos de pulmões, sendo 45 com lesões macroscópicas (CLM) e 75 sem lesões macroscópicas (SLM) foram analisados pela histopatologia. Destes, foram selecionadas aleatoriamente 32 amostras (sendo 16 sem lesões macroscópicas e 16 com lesões macroscópicas) para a realização dos testes de imunoistoquímica (para a detecção do Mycoplasma hyopneumoniae) e qPCR (para detecção do PCV2). Não foi observada diferença significativa entre os grupos (SLM e CLM) em nenhum dos testes, o que indica que a ausência de lesões macroscópicas não descarta a possibilidade da presença dos agentes nem de lesões microscópicas. A técnica de histopatologia representou uma importante ferramenta para se aproximar ao diagnóstico. Embora as lesões não sejam patognomônicas para os agentes avaliados, elas são bastante sugestivas dos quadros viral e bacteriano, e esses achados, aliados a presença de sinais clínicos e detecção do agente (que são mais frequentemente realizados por qPCR e imunoistoquímica), confirmam o diagnóstico de cada agente isoladamente ou da coinfecção. Apesar das amostras analisadas pelo qPCR terem apresentado grande número de resultados negativos e baixa carga viral, diversos autores relatam que o pulmão é um dos órgãos com menor carga viral de PCV2, sendo desta forma os órgãos linfóides, principais alvos do PCV2, mais indicados para a quantificação viral. Foi verificada uma correlação positiva do teste de imunoistoquímica com o teste histopatológico de acordo com o teste estatístico de Pearson, o que confirma a presença do agente nas lesões observadas na histopatologia. Estudos futuros utilizando a coleta de linfonodos (órgão alvo do PCV2) e amostras coletadas de diferentes regiões anatômicas pulmonares devem ser realizados para avaliar a co-infecção de uma forma mais acurada.

Histopathological findings in 25 pig tissue samples, which indicated PCVD (porcine circovirus diseases), were studied. Pig tissue samples originated from 5 different pig-farms in the north-west part of Croatia. Histopathological lesions showed two clinical pictures of the disease: porcine multisystemic wasting syndrome (PMWS) and porcine dermatitis nephropathy syndrome (PDNS). All samples were tested by PCR for the presence of porcine circovirus type 2 (PCV2). Twenty of them were PCV2 positive. PCV2 DNA was quantified by realtime-PCR in all twenty samples with a wide range of PCV2 loads ranging from 105 to 1010 PCV2 genomes/μL. The highest PCV2 loads were detected in pigs with PMWS lesions, while PDNS affected pigs had lower PCV2 loads. The 20 PCV2 isolates were sequenced and analyzed. Nineteen isolates belonged to PCV2 group 1 and only one to PCV2 group 2. We found no link between genotypes and clinical form of the disease., U ovom su radu izneseni patohistološki rezultati 25 uginulih svinja s pet različitih svinjogojskih farmi na području sjeverozapadne Hrvatske, koje su uginule pod znakovima sindroma kržljavosti odbijene prasadi i sindroma nekrotičnoga dermatitisa. Klinički i patohistološki nalazi upućivali su na cirkovirusnu infekciju svinja. Lančanom reakcijom polimerazom dokazana je virusna DNK svinjskog cirkovirusa tipa 2 u dvadeset tkivnih uzoraka. Ti su uzorci nadalje bili pretraženi lančanom reakcijom polimerazom u stvarnom vremenu kako bi se ustanovila količina virusne DNK. U svim uzorcima ustanovljena je značajna količina virusnoga genoma, koja je iznosila između 105 i 1010 kopija genoma/μL. Veća količina virusa dokazana je u onim uzorcima koji su potjecali od svinja uginulih pod znakovima sindroma kržljavosti, a nešto manja količina virusa dokazana je u svinja koje su bolovale od sindroma nekrotičnoga dermatitisa. Svi PCR-proizvodi sekvencirani su kako bi se ustanovila pripadnost određenom genotipu svinjskoga cirkovirusa tipa 2 (SCV-2). Devetnaest izolata od dvadeset sekvenciranih, genotipski pripada skupini 1 SCV-2, a samo jedan izolat skupini 2 SCV-2. Takva genotipska pripadnost podudarna je s podatcima iz drugih europskih zemalja, gdje prevladava genotip 1 SCV-2 od 2003. god., a prije toga je dominirao genotip 2 SCV-2 kao etiološki čimbenik cirkovirusnih bolesti svinja.

Histopathological findings in 25 pig tissue samples, which indicated PCVD (porcine circovirus diseases)were studied. All samples were tested by PCR for the presence of porcine circovirus type 2(PCV2). Twenty of them were were PCV-2 positive and after that were sequenced and analyzed. Nineteen isolates belonged to PCV2 group 1 and only one to PCV2 group 2.

The occurence of "porcine circovirus associated disesases" in pigs in Croatia has been described. This report is aimed to provide information on cocirculation of two genotypes of PCV2 in Croatia. Moreover, it is documented that PCV2 was associated with clinical and pathomorphological canges similar to classical swine fever.

Porcine circovirus type 2 (PCV2) is the primary causative agent for post-weaning, multisystemic, wasting syndrome. Consequently, serologic detection of and vaccination against PCV2 are important for the swine industry. Among several serological tests, the enzyme-linked immunosorbent assay (ELISA) is commonly used to measure anti-PCV2 antibody levels. In the present study, we used two commercial ELISA systems to comparatively evaluate anti-PCV2 antibodies in field pigs treated with three different PCV2 vaccines. Among a total of 517 serum samples, the results of the two ELISAs were fully concordant for 365 positive and 42 negative samples, indicating 78.7% agreement. In addition, the Pearson coefficient (0.636) indicated a moderate correlation between data from the two ELISAs. Results from the farms with pigs vaccinated with the three different PCV2 vaccines demonstrated that most of the vaccinated animals underwent seroconversion. However, the increase and duration of antibody titers varied depending on the vaccine, the presence of maternal antibodies, and the vaccination program. PCV2 serologic status and anti-PCV2 antibody levels of herds from this study could be utilized to determine the best timing for vaccination and assessing vaccination compliance.