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Background: Porcine Circovirus type 2 (PCV2) infections are distributed worldwide and cause Porcine Circovirus Associated Disease (PCVAD). To minimize the impact of PCV2 infection on swine health and production, different vaccination schemes have been used since 2006. However, the association between vaccination schemes, virus load and disease under field conditions are not completely understood. Therefore, the objective of this study was to compare the effect of two different PCV2 vaccination schemes on the humoral response and PCV2 load in pigs after weaning under field conditions. Methods: Two commercial pig farms (Farm A and B), endemically infected with PCV2, which were using two different PCV2 subunit vaccinations schemes for sow, gilts and piglets, were selected. We designed a longitudinal study and measured IgG levels by ELISA and virus load by quantitative PCR in pigs after weaning. Forty 3-week old piglets were randomly selected at weaning and followed for 20 weeks. IgG levels and virus loads were compared within and between farms and considered statistically different if the non-parametric Kruskal Wallis test p value was lower than 0.05. Results: We found that low virus loads were maintained in pigs from both farms regardless of the vaccination scheme used (p>0.05). However, there was significant difference in the mean IgG levels observed over time (p<0.05), suggesting that different humoral immune response are not necessarily associated with different virus loads observed over time. Conclusions: These results are important because they can help to prevent PCV2 infections using different vaccination schemes to minimize the effect of PCVAD on swine health and production.

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Abstract Economic losses with high mortality rate associated with Porcine circovirus type 2 (PCV2) is reported worldwide. PCV2 commercial vaccine was introduced in 2006 in U.S. and in 2008 in Brazil. Although PCV2 vaccines have been widely used, cases of PCV2 systemic disease have been reported in the last years. Eleven nursery or fattening pigs suffering from PCV2 systemic disease were selected from eight PCV2-vaccinated farms with historical records of PCV2 systemic disease in Southern Brazil. PCV2 genomes were amplified and sequenced from lymph node samples of selected pigs. The comparison among the ORF2 amino acid sequences of PCV2 isolates revealed three amino acid substitutions in the positions F57I, N178S and A190T, respectively. Using molecular modeling, a structural model for the capsid protein of PCV2 was built. Afterwards, the mutated residues positions were identified in the model. The structural analysis of the mutated residues showed that the external residue 190 is close to an important predicted region for antibodies recognition. Therefore, changes in the viral protein conformation might lead to an inefficient antibody binding and this could be a relevant mechanism underlying the recent vaccine failures observed in swine farms in Brazil.

Assuntos

RESUMO

Economic losses with high mortality rate associated with Porcine circovirus type 2 (PCV2) is reported worldwide. PCV2 commercial vaccine was introduced in 2006 in U.S. and in 2008 in Brazil. Although PCV2 vaccines have been widely used, cases of PCV2 systemic disease have been reported in the last years. Eleven nursery or fattening pigs suffering from PCV2 systemic disease were selected from eight PCV2-vaccinated farms with historical records of PCV2 systemic disease in Southern Brazil. PCV2 genomes were amplified and sequenced from lymph node samples of selected pigs. The comparison among the ORF2 amino acid sequences of PCV2 isolates revealed three amino acid substitutions in the positions F57I, N178S and A190T, respectively. Using molecular modeling, a structural model for the capsid protein of PCV2 was built. Afterwards, the mutated residues positions were identified in the model. The structural analysis of the mutated residues showed that the external residue 190 is close to an important predicted region for antibodies recognition. Therefore, changes in the viral protein conformation might lead to an inefficient antibody binding and this could be a relevant mechanism underlying the recent vaccine failures observed in swine farms in Brazil.(AU)

Assuntos

RESUMO

Economic losses with high mortality rate associated with Porcine circovirus type 2 (PCV2) is reported worldwide. PCV2 commercial vaccine was introduced in 2006 in U.S. and in 2008 in Brazil. Although PCV2 vaccines have been widely used, cases of PCV2 systemic disease have been reported in the last years. Eleven nursery or fattening pigs suffering from PCV2 systemic disease were selected from eight PCV2-vaccinated farms with historical records of PCV2 systemic disease in Southern Brazil. PCV2 genomes were amplified and sequenced from lymph node samples of selected pigs. The comparison among the ORF2 amino acid sequences of PCV2 isolates revealed three amino acid substitutions in the positions F57I, N178S and A190T, respectively. Using molecular modeling, a structural model for the capsid protein of PCV2 was built. Afterwards, the mutated residues positions were identified in the model. The structural analysis of the mutated residues showed that the external residue 190 is close to an important predicted region for antibodies recognition. Therefore, changes in the viral protein conformation might lead to an inefficient antibody binding and this could be a relevant mechanism underlying the recent vaccine failures observed in swine farms in Brazil.

Assuntos

RESUMO

Abstract Economic losses with high mortality rate associated with Porcine circovirus type 2 (PCV2) is reported worldwide. PCV2 commercial vaccine was introduced in 2006 in U.S. and in 2008 in Brazil. Although PCV2 vaccines have been widely used, cases of PCV2 systemic disease have been reported in the last years. Eleven nursery or fattening pigs suffering from PCV2 systemic disease were selected from eight PCV2-vaccinated farms with historical records of PCV2 systemic disease in Southern Brazil. PCV2 genomes were amplified and sequenced from lymph node samples of selected pigs. The comparison among the ORF2 amino acid sequences of PCV2 isolates revealed three amino acid substitutions in the positions F57I, N178S and A190T, respectively. Using molecular modeling, a structural model for the capsid protein of PCV2 was built. Afterwards, the mutated residues positions were identified in the model. The structural analysis of the mutated residues showed that the external residue 190 is close to an important predicted region for antibodies recognition. Therefore, changes in the viral protein conformation might lead to an inefficient antibody binding and this could be a relevant mechanism underlying the recent vaccine failures observed in swine farms in Brazil.

RESUMO

Since its discovery, Porcine circovirus type 2 has emerged as one of the most relevant swine infectious diseases, causing relevant economic losses for the pig industry. While four genotypes were identified, only three (PCV2a, PCV2b and PCV2d) are currently circulating and display a worldwide distribution. Another genotype, PCV2c, has been described only once in Danish archive samples collected between 1980 and 1990. In addition to commercial pigs, PCV2 has been demonstrated to infect wild boars and other wild species, which can potentially serve as a reservoir for domestic populations. In this study, eight sequences obtained from feral pigs in the Pantanal region (Mato Grosso do Sul State, Brazil) were compared with reference sequences and other Brazilian sequences, and the results revealed remarkable genetic diversity, with all four genotypes currently recognised being detected (PCV2a, PCV2b, PCV2c and PCV2d). This finding represents a remarkable discovery, as it is the first detection of PCV2c since 1990 and the first-ever detection of PCV2c in live animals. The peculiar population history and ecological scenario of feral pigs in the Pantanal coupled with the complex, and still only partially known relationship of feral pigs with other PCV2 susceptible species (i.e., domestic pigs, wild boars and peccaries), open exciting questions concerning PCV2 origin and evolution. Overall, the results of the present study led us to form the following hypothesis: the PCV2 strains found in feral pigs may be the last descent of the strains that circulated among European pigs in the past, or they may have infected these feral pigs more recently through a bridge species.

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Investigou-se a presença de sequências genômicas do circovírus suíno tipo 2 (PCV2) e do parvovírus suíno (PPV) em 147 fetos suínos natimortos e mumificados. Estas amostras, provenientes de 39 granjas localizadas em oito estados brasileiros, foram coletadas entre os anos de 2006 e 2008. Utilizaram-se fragmentos de coração e pulmão para extração do DNA total e posterior amplificação de fragmentos correspondentes aos patógenos virais pela técnica de reação em cadeia da polimerase (PCR). Entre as 147 amostras, 74 (50,3%) foram positivas ao PCV2 enquanto nove amostras (6,2%) apresentaram coinfecção com o PCV2 e o PPV. Nenhuma amostra foi positiva apenas para PPV. Entre as 39 granjas estudadas, 21 (53,8%) apresentaram fetos positivos ao PCV2, sendo detectada coinfecção com o PCV2 e o PPV em três (7,7%). Esses resultados indicam que o PCV2 pode ser um importante agente infeccioso causador de morte embrionária e fetal em suínos no Brasil e deve ser incluído na lista de diagnóstico diferencial.(AU)

This study investigated the presence of genome sequences of the porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV) in 147 porcine stillbirths and mummified fetuses. These samples, originated from 39 farms located in eight Brazilian states, were collected between 2006 and 2008. Heart and lung fragments were used for extraction of total DNA and later amplification of correspondent fragments of the virus pathogens through polymerase chain reaction (PCR) technique. Out of 147 samples, 74 (50.3%) were positive for PCV2 while nine samples (6.2%) were positive for PCV2 and PPV. None of the samples were positive just for PPV. Out of 39 investigated farms, 21 (53.8%) had fetuses positive for PCV2 while co-infection with PCV2 and PPV was detected in 3 farms (7.7%). These results indicate that PCV2 could be an important infection agent in cases of porcine stillbirths and mummified fetuses in Brazil and must be included at differential diagnostic list.(AU)

Assuntos

RESUMO

Investigou-se a presença de sequências genômicas do circovírus suíno tipo 2 (PCV2) e do parvovírus suíno (PPV) em 147 fetos suínos natimortos e mumificados. Estas amostras, provenientes de 39 granjas localizadas em oito estados brasileiros, foram coletadas entre os anos de 2006 e 2008. Utilizaram-se fragmentos de coração e pulmão para extração do DNA total e posterior amplificação de fragmentos correspondentes aos patógenos virais pela técnica de reação em cadeia da polimerase (PCR). Entre as 147 amostras, 74 (50,3%) foram positivas ao PCV2 enquanto nove amostras (6,2%) apresentaram coinfecção com o PCV2 e o PPV. Nenhuma amostra foi positiva apenas para PPV. Entre as 39 granjas estudadas, 21 (53,8%) apresentaram fetos positivos ao PCV2, sendo detectada coinfecção com o PCV2 e o PPV em três (7,7%). Esses resultados indicam que o PCV2 pode ser um importante agente infeccioso causador de morte embrionária e fetal em suínos no Brasil e deve ser incluído na lista de diagnóstico diferencial.

This study investigated the presence of genome sequences of the porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV) in 147 porcine stillbirths and mummified fetuses. These samples, originated from 39 farms located in eight Brazilian states, were collected between 2006 and 2008. Heart and lung fragments were used for extraction of total DNA and later amplification of correspondent fragments of the virus pathogens through polymerase chain reaction (PCR) technique. Out of 147 samples, 74 (50.3%) were positive for PCV2 while nine samples (6.2%) were positive for PCV2 and PPV. None of the samples were positive just for PPV. Out of 39 investigated farms, 21 (53.8%) had fetuses positive for PCV2 while co-infection with PCV2 and PPV was detected in 3 farms (7.7%). These results indicate that PCV2 could be an important infection agent in cases of porcine stillbirths and mummified fetuses in Brazil and must be included at differential diagnostic list.

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RESUMO

Postweaning multisystemic wasting syndrome (PMWS) is of economical importance a disease caused by porcine circovirus type 2 (PCV2). A retrospective investigation was performed on paraffin-embedded organs samples from swine submitted to Embrapa Swine and Poultry Research Center between 1985 and 1998 for histopathologic diagnosis. A total of 25 cases were chosen from the archival collection of the Animal Health Laboratory at Embrapa Swine and Poultry based on characteristic pathological lesions of PMWS, such as lymphadenopathy, interstitial pneumonia, hepatitis and interstitial nephritis. The sections were investigated by nested-PCR (polymerase chain reaction) which used specific primers for the ORF2 sequence of the PCV2 and by immunohistochemistry using a monoclonal antibody specific for PCV2 capsid antigen. Virus specific DNA and antigen were detected in tissue samples of two out of 25 analyzed cases. The earliest positive sample originated from 1988. These results indicate that PCV2 is present in Brazil since 1988.

A síndrome multissistêmica do definhamento dos suínos (SMDS) é uma doença de importância econômica causada pelo circovírus suíno tipo 2 (PCV2). Uma pesquisa retrospectiva foi realizada em amostras de órgãos de suínos fixados em blocos de parafina arquivados, que haviam sido submetidos à Embrapa Suínos e Aves entre 1985 e 1998 para o diagnóstico histopatológico. Vinte e cinco casos foram selecionados com base nas lesões histológicas características da SMDS, tais como linfoadenopatia, pneumonia intersticial, hepatite e nefrite intersticial. A presença de PCV2 nos cortes histológicos foi pesquisada por reação em cadeia da polimerase interna (nested-PCR), na qual utilizou-se primers específicos para a seqüência da ORF2 do PCV2 e também por imunoistoquímica, utilizando um anticorpo monoclonal específico para o capsídeo do PCV2. O DNA viral e os antígenos específicos do PCV2 foram detectados em amostras de tecidos de dois dos 25 casos analisados, sendo um desses datado de 1988. Esses resultados indicam que o PCV2 já estava presente no Brasil desde 1988.

RESUMO

Postweaning multisystemic wasting syndrome (PMWS) is of economical importance a disease caused by porcine circovirus type 2 (PCV2). A retrospective investigation was performed on paraffin-embedded organs samples from swine submitted to Embrapa Swine and Poultry Research Center between 1985 and 1998 for histopathologic diagnosis. A total of 25 cases were chosen from the archival collection of the Animal Health Laboratory at Embrapa Swine and Poultry based on characteristic pathological lesions of PMWS, such as lymphadenopathy, interstitial pneumonia, hepatitis and interstitial nephritis. The sections were investigated by nested-PCR (polymerase chain reaction) which used specific primers for the ORF2 sequence of the PCV2 and by immunohistochemistry using a monoclonal antibody specific for PCV2 capsid antigen. Virus specific DNA and antigen were detected in tissue samples of two out of 25 analyzed cases. The earliest positive sample originated from 1988. These results indicate that PCV2 is present in Brazil since 1988.

A síndrome multissistêmica do definhamento dos suínos (SMDS) é uma doença de importância econômica causada pelo circovírus suíno tipo 2 (PCV2). Uma pesquisa retrospectiva foi realizada em amostras de órgãos de suínos fixados em blocos de parafina arquivados, que haviam sido submetidos à Embrapa Suínos e Aves entre 1985 e 1998 para o diagnóstico histopatológico. Vinte e cinco casos foram selecionados com base nas lesões histológicas características da SMDS, tais como linfoadenopatia, pneumonia intersticial, hepatite e nefrite intersticial. A presença de PCV2 nos cortes histológicos foi pesquisada por reação em cadeia da polimerase interna (nested-PCR), na qual utilizou-se primers específicos para a seqüência da ORF2 do PCV2 e também por imunoistoquímica, utilizando um anticorpo monoclonal específico para o capsídeo do PCV2. O DNA viral e os antígenos específicos do PCV2 foram detectados em amostras de tecidos de dois dos 25 casos analisados, sendo um desses datado de 1988. Esses resultados indicam que o PCV2 já estava presente no Brasil desde 1988.