RESUMO
Backyard pig populations are not monitored for influenza A virus (IAV) in Brazil and there are limited data about seroprevalence and risk factors in these populations. Our goal was to assess possible factors associated with IAV seroprevalence in backyard pig populations using an indirect ELISA protocol based on a recombinant nucleoprotein. Following the IAV screening using NP-ELISA, subtype-specific serology based on hemagglutination inhibition (HI) assay of the ELISA-positive pigs was conducted. The survey comprised a total of 1,667 sera samples collected in 2012 and 2014 in 479 holdings and the estimated seroprevalence was 5.3% (3.84%-7.33%) and 2.3% (1.34%-3.71%) in the respective years. In both years, H1N1pdm09 was the most prevalent subtype. The multivariable analysis showed main factors such as "age," "sex," "number of suckling pigs" and "neighbours raising pigs" that presented the greatest effect on IAV seroprevalence in these pig populations. These factors may be associated with the low biosecurity measures and management of backyard holdings. In addition, the low IAV seroprevalences found in these backyard pig populations could be related to a low number of animals in each pig holding and low animal movement/replacement that do not favour IAV transmission dynamics. This low frequency of H1N1pdm09 seropositive pigs could also be due to sporadic human-to-pig transmission of what is now a human seasonal influenza A virus; however, these factors should be explored in future studies. Herein, these results highlight the importance of IAV continued surveillance in backyard pig holdings, since it is poorly known which IAVs are circulating in these populations and the risk they could pose to public health and virus transmission to commercial farms.
Assuntos
Vírus da Influenza A/isolamento & purificação , Infecções por Orthomyxoviridae/veterinária , Doenças dos Suínos/virologia , Animais , Brasil/epidemiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Masculino , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/epidemiologiaRESUMO
INTRODUCTION AND OBJECTIVE: Recently, investigations in a swine herd identified evidence of the existence of a novel member of the Hepadnavirus family endemic in swine. The aim of this study was to investigate the serological and molecular markers of Hepadnavirus circulation in Brazilian domestic swine and wild boar herds, and to evaluate the identity with HBV and other Hepadnaviruses reported previously. MATERIALS AND METHODS: For the study, 376 swine were screened for hepatitis B virus serological markers. Analyses were performed in serum samples using commercial enzyme-linked immunosorbent assay (ELISA) kits (DiaSorin®) for anti-HBc, HBsAg and anti-HBs. Reactive and undetermined swine serum samples were selected to perform DNA viral extraction (QIAamp DNA Mini Kit, Qiagen®), partial genome amplification and genome sequencing. RESULTS: From 376 swine samples analysed, 28 (7.45%) were reactive to anti-HBc, 3 (0.80%) to HBsAg and 6 (1.6%) to anti-HBs. Besides, more 17 (4.52%) swine samples analyzed were classified in the grey zone of the EIA test to anti-HBc and 2 (0.53%) to HBsAg. From 49 samples molecularly analyzed after serological trial, 4 samples showed a positive result for the qualitative PCR for Hepadnavirus. Phylogenetic reconstruction using partial genome sequencing (360 bp) of 3 samples showed similarity with HBV with 90.8-96.3% of identity. CONCLUSIONS: Serological and molecular data showed evidence of the circulation of a virus similar to hepatitis B virus in swine.
Assuntos
Vírus da Hepatite B/classificação , Vírus da Hepatite B/isolamento & purificação , Hepatite B/veterinária , Doenças dos Suínos/epidemiologia , Animais , Brasil/epidemiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Hepatite B/epidemiologia , Hepatite B/virologia , Anticorpos Anti-Hepatite B/análise , Antígenos de Superfície da Hepatite B/análise , Vírus da Hepatite B/genética , Masculino , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/veterinária , Análise de Sequência de DNA/veterinária , Suínos , Doenças dos Suínos/virologia , Proteínas Virais/genéticaRESUMO
BACKGROUND: Swine influenza virus (SIV) is the cause of an acute respiratory disease that affects swine worldwide. In Brazil, SIV has been identified in pigs since 1978. After the emergence of pandemic H1N1 in 2009 (H1N1pdm09), few studies reported the presence of influenza virus in Brazilian herds. OBJECTIVES: The objective of this study was to evaluate the serological profile for influenza virus in farrow-to-finish pig farms in Minas Gerais state, Brazil. METHODS: Thirty farms with no SIV vaccination history were selected from the four larger pig production areas in Minas Gerais state (Zona da Mata, Triângulo Mineiro/Alto Paranaíba, South/Southwest and the Belo Horizonte metropolitan area). At each farm, blood samples were randomly collected from 20 animals in each production cycle category: breeding animals (sows and gilts), farrowing crate (2-3 weeks), nursery (4-7 weeks), grower pigs (8-14 weeks), and finishing pigs (15-16 weeks), with 100 samples per farm and a total of 3000 animals in this study. The samples were tested for hemagglutination inhibition activity against H1N1 pandemic strain (A/swine/Brazil/11/2009) and H3N2 SIV (A/swine/Iowa/8548-2/98) reference strain. RESULTS: The percentages of seropositive animals for H1N1pdm09 and H3N2 were 26.23% and 1.57%, respectively, and the percentages of seropositive herds for both viruses were 96.6% and 13.2%, respectively. CONCLUSIONS: The serological profiles differed for both viruses and among the studied areas, suggesting a high variety of virus circulation around the state, as well as the presence of seronegative animals susceptible to influenza infection and, consequently, new respiratory disease outbreaks.
Assuntos
Anticorpos Antivirais/sangue , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Infecções por Orthomyxoviridae/veterinária , Doenças dos Suínos/epidemiologia , Agricultura , Animais , Brasil , Testes de Inibição da Hemaglutinação , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/prevenção & controle , Doenças dos Suínos/virologiaRESUMO
Influenza is a viral disease that affects human and several animal species. In Brazil, H1N1, H3N2 and 2009 pandemic H1N1 A(H1N1)pdm09 influenza A viruses (IAV) circulate in domestic swine herds. Wild boars are also susceptible to IAV infection but in Brazil until this moment there are no reports of IAV infection in wild boars or in captive wild boars populations. Herein the occurrence of IAV in captive wild boars with the presence of lung consolidation lesions during slaughter was investigated. Lung samples were screened by RT-PCR for IAV detection. IAV positive samples were further analyzed by quantitative real-time PCR (qRRT-PCR), virus isolation, genomic sequencing, histopathology and immunohistochemistry (IHC). Eleven out of 60 lungs (18.3%) were positive for IAV by RT-PCR and seven out of the eleven were also positive for A(H1N1)pdm09 by qRRT-PCR. Chronic diffuse bronchopneumonia was observed in all samples and IHC analysis was negative for influenza A antigen. Full genes segments of H1N2 IAV were sequenced using Illumina's genome analyzer platform (MiSeq). The genomic analysis revealed that the HA and NA genes clustered with IAVs of the human lineage and the six internal genes were derived from the H1N1pdm09 IAV. This is the first report of a reassortant human-like H1N2 influenza virus infection in captive wild boars in Brazil and indicates the need to monitor IAV evolution in Suidae populations.
Assuntos
Genoma Viral/genética , Vírus da Influenza A Subtipo H1N2/classificação , Vírus da Influenza A Subtipo H1N2/genética , Infecções por Orthomyxoviridae/veterinária , Infecções por Orthomyxoviridae/virologia , Filogenia , Sus scrofa/virologia , Doenças dos Suínos/virologia , Animais , Brasil , Genes Virais/genética , Humanos , Vírus da Influenza A Subtipo H1N2/isolamento & purificação , Pulmão/virologia , Dados de Sequência Molecular , SuínosRESUMO
Este trabalho descreve a colheita adequada de amostras, as técnicas/procedimentos disponíveis para o diagnóstico de influenza A em suínos, assim como os resultados e suas respectivas interpretações, para auxiliar médicos veterinários de campo na identificação dessa doença. Em suínos vivos, as amostras adequadas são: secreção nasal, fluido oral e sangue (soro). Para suínos mortos, colher preferencialmente amostras de pulmão com consolidação cranioventral. Secreção nasal e fragmentos de pulmão refrigerado são utilizados para detectar partícula viral viável (isolamento viral - IV) ou ácido nucleico viral (RT-PCR convencional e RT-PCR em tempo real). As amostras não devem ser congeladas, pois o vírus é inativado a -20°C. A caracterização molecular dos isolados é feita pela análise filogenética obtida pelo sequenciamento de DNA. O soro é utilizado para a detecção de anticorpos (Acs) por meio do teste da inibição da hemaglutinação e ELISA. O fluido oral pode ser utilizado para detecção de anticorpo (ELISA) ou de vírus. Fragmentos de pulmão fixados em formol a 10% são examinados microscopicamente para identificar pneumonia broncointersticial e para detecção de antígeno viral pela imuno-histoquímica (IHQ). Para o sucesso do diagnóstico, as amostras devem ser colhidas de suínos que estão preferencialmente na fase aguda da doença, para aumentar as chances de detecção viral. As melhores opções para o diagnóstico de influenza A em suínos vivos são RT-PCR e isolamento viral de amostras de swab nasal ou fluido oral. Pulmão para análise por RT-PCR, isolamento viral ou IHQ é a amostra de escolha em suínos mortos. Testes sorológicos têm valor diagnóstico limitado e são utilizados apenas para determinar o estado imune do rebanho, não indicando doença clínica, pois os Acs são detectados 7-10 dias pós-infecção (fase subaguda). O diagnóstico de influenza é importante para avaliar o envolvimento desse agente no complexo de doença respiratória suína. Além disso, o isolamento do vírus influenza é essencial para o monitoramento dos principais subtipos circulantes em uma determinada região ou país, assim como para a detecção de novos rearranjos virais, já que influenza é considerada uma zoonose.(AU)
This article is intended to describe the adequate sample collection, the laboratory procedures/techniques, the expected results and their interpretation for diagnosis of influenza infection in swine, serving as a support for field veterinarians. In live pigs, the samples to be taken are nasal secretions, oral fluids and blood. For dead pigs, preference should be given to samples of cranioventral lung consolidation. Nasal discharge and chilled lung fragments are used for detection of virus (virus isolation - VI) or viral nucleic acids (conventional RT-PCR and real-time RT-PCR). Samples should not be frozen, because the virus is inactivated at -20°C. Molecular characterization of isolates is performed by phylogenetic analysis of gene sequences obtained by DNA sequencing. Serum is used for the detection of antibodies using hemagglutination inhibition (HI) test and ELISA. Oral fluid may be used for either antibody (ELISA) or viral detection. Fragments of lung fixed in 10% formaldehyde are used for histopathological analysis to identify bronchointerstitial pneumonia, and for immunohistochemistry (IHC) for antigens. For a successful diagnosis, sampling should be preferably performed in the acute phase of the disease to improve chances of virus detection. The best options to perform the diagnosis of influenza A in a swine herd are RT-PCR and VI from nasal swabs or oral fluid in live pigs and/or lung tissue for RT-PCR, VI or IHC in dead pigs. Serological tests are of very limited diagnostic value and are useful only to determine the immune status of the herd, not indicating clinical disease, because antibodies are detected after 7-10 days post infection (subacute phase). The diagnosis of influenza is important to evaluate the involvement of this agent in the complex of respiratory diseases in pigs. Furthermore, the isolation of influenza virus is essential for monitoring the main subtypes circulating in a given region or country, as well as for the detection of potential new viral reassortants, because influenza is considered a zoonosis.(AU)
Assuntos
Animais , Alphainfluenzavirus/isolamento & purificação , Manejo de Espécimes , Suínos/virologia , Técnicas e Procedimentos Diagnósticos/veterinária , Saliva , Reação em Cadeia da PolimeraseRESUMO
Este trabalho descreve a colheita adequada de amostras, as técnicas/procedimentos disponíveis para o diagnóstico de influenza A em suínos, assim como os resultados e suas respectivas interpretações, para auxiliar médicos veterinários de campo na identificação dessa doença. Em suínos vivos, as amostras adequadas são: secreção nasal, fluido oral e sangue (soro). Para suínos mortos, colher preferencialmente amostras de pulmão com consolidação cranioventral. Secreção nasal e fragmentos de pulmão refrigerado são utilizados para detectar partícula viral viável (isolamento viral - IV) ou ácido nucleico viral (RT-PCR convencional e RT-PCR em tempo real). As amostras não devem ser congeladas, pois o vírus é inativado a -20°C. A caracterização molecular dos isolados é feita pela análise filogenética obtida pelo sequenciamento de DNA. O soro é utilizado para a detecção de anticorpos (Acs) por meio do teste da inibição da hemaglutinação e ELISA. O fluido oral pode ser utilizado para detecção de anticorpo (ELISA) ou de vírus. Fragmentos de pulmão fixados em formol a 10% são examinados microscopicamente para identificar pneumonia broncointersticial e para detecção de antígeno viral pela imuno-histoquímica (IHQ). Para o sucesso do diagnóstico, as amostras devem ser colhidas de suínos que estão preferencialmente na fase aguda da doença, para aumentar as chances de detecção viral. As melhores opções para o diagnóstico de influenza A em suínos vivos são RT-PCR e isolamento viral de amostras de swab nasal ou fluido oral. Pulmão para análise por RT-PCR, isolamento viral ou IHQ é a amostra de escolha em suínos mortos. Testes sorológicos têm valor diagnóstico limitado e são utilizados apenas para determinar o estado imune do rebanho, não indicando doença clínica, pois os Acs são detectados 7-10 dias pós-infecção (fase subaguda). O diagnóstico de influenza é importante para avaliar o envolvimento desse agente no complexo de doença respiratória suína. Além disso, o isolamento do vírus influenza é essencial para o monitoramento dos principais subtipos circulantes em uma determinada região ou país, assim como para a detecção de novos rearranjos virais, já que influenza é considerada uma zoonose.
This article is intended to describe the adequate sample collection, the laboratory procedures/techniques, the expected results and their interpretation for diagnosis of influenza infection in swine, serving as a support for field veterinarians. In live pigs, the samples to be taken are nasal secretions, oral fluids and blood. For dead pigs, preference should be given to samples of cranioventral lung consolidation. Nasal discharge and chilled lung fragments are used for detection of virus (virus isolation - VI) or viral nucleic acids (conventional RT-PCR and real-time RT-PCR). Samples should not be frozen, because the virus is inactivated at -20°C. Molecular characterization of isolates is performed by phylogenetic analysis of gene sequences obtained by DNA sequencing. Serum is used for the detection of antibodies using hemagglutination inhibition (HI) test and ELISA. Oral fluid may be used for either antibody (ELISA) or viral detection. Fragments of lung fixed in 10% formaldehyde are used for histopathological analysis to identify bronchointerstitial pneumonia, and for immunohistochemistry (IHC) for antigens. For a successful diagnosis, sampling should be preferably performed in the acute phase of the disease to improve chances of virus detection. The best options to perform the diagnosis of influenza A in a swine herd are RT-PCR and VI from nasal swabs or oral fluid in live pigs and/or lung tissue for RT-PCR, VI or IHC in dead pigs. Serological tests are of very limited diagnostic value and are useful only to determine the immune status of the herd, not indicating clinical disease, because antibodies are detected after 7-10 days post infection (subacute phase). The diagnosis of influenza is important to evaluate the involvement of this agent in the complex of respiratory diseases in pigs. Furthermore, the isolation of influenza virus is essential for monitoring the main subtypes circulating in a given region or country, as well as for the detection of potential new viral reassortants, because influenza is considered a zoonosis.
Assuntos
Animais , Alphainfluenzavirus/isolamento & purificação , Manejo de Espécimes , Suínos/virologia , Técnicas e Procedimentos Diagnósticos/veterinária , Reação em Cadeia da Polimerase , SalivaRESUMO
The aim of this work was to detect serum antibodies specific to influenza viruses in swine in Brazil. Serum samples of 355 pigs from 17 herds in Minas Gerais state were tested by hemagglutination inhibition (HI) for antibodies against H1N1 swine (SIV) and human influenza viruses, and H3N2 SIV. HI revealed that 158 animals (44·5%) and 11 herds (64·7%) were positive for H1N1 SIV, 36 animals (10·1%) and four herds (23·5%) were positive for H3N2 SIV, and 136 animals (38·3%) and 10 herds (58·8%) were positive for H1N1 human. This study indicates that swine influenza is disseminated throughout Minas Gerais state, Brazil.
Assuntos
Anticorpos Antivirais/sangue , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Infecções por Orthomyxoviridae/veterinária , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/virologia , Animais , Brasil/epidemiologia , Testes de Inibição da Hemaglutinação , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Estudos Soroepidemiológicos , SuínosRESUMO
Influenza A viruses cause acute respiratory disease in swine. Viruses with H1 hemagglutinin genes from the human seasonal lineage (δ-cluster) have been isolated from North American swine since 2003. The objective of this work was to study the pathogenesis and transmission of δ-cluster H1 influenza viruses in swine, comparing three isolates from different phylogenetic subclusters, geographic locations, and years of isolation. Two isolates from the δ2 subcluster, A/sw/MN/07002083/07 H1N1 (MN07) and A/sw/IL/00685/05 H1N1 (IL05), and A/sw/TX/01976/08 H1N2 (TX08) from the δ1 sub-cluster were evaluated. All isolates caused disease and were transmitted to contact pigs. Respiratory disease was apparent in pigs infected with MN07 and IL05 viruses; however, clinical signs and lung lesions were reduced in severity as compared to TX08. On day 5 following infection MN07-infected pigs had lower virus titers than the TX08 pigs, suggesting that although this H1N1 was successfully transmitted, it may not replicate as efficiently in the upper or lower respiratory tract. MN07 and IL05 H1N1 induced higher serum antibody titers than TX08. Greater serological cross-reactivity was observed for viruses from the same HA phylogenetic sub-cluster; however, antigenic differences between the sub-clusters may have implications for disease control strategies for pigs.
RESUMO
This study evaluated histological lesions in kidney samples from pigs with nephritis in two slaughterhouses in the State of Mato Grosso, Brazil. Four hundred samples were subjected to histology, anti-porcine circovirus type 2 (PCV2) immunohistochemistry (IHC), anti-Leptospira sp. immunofluorescence (IF), and polymerase chain reaction (PCR) for PCV2, porcine parvovirus (PPV), and Torque teno virus type 1 and 2 (TTV1, TTV2) detection. Histological lesions were found in 81% of the samples, and mononuclear interstitial nephritis was the most frequent lesion (77.50%). A follicular pattern was observed in 40.97% of the interstitial nephritis lesions. PCV2, PPV, TTV1, and TTV2 were identified in the kidneys by PCR in 27.25%, 28.50%, 94%, and 87.5% of the samples, respectively. Leptospira sp. was not detected through IF. Infection by PCV2 (PCR) and the presence of histological lesions (P=0.008) and giant cells (P=0.0016) were significantly associated. An association was observed between the TTV2-TTV1 co-infection (P<0.0001) and the risk for pathogenesis. These findings indicated that PCV2, PPV, TTV1, and TTV2 were widely distributed among pigs in the local farms and that the presence of these agents should be considered in the differential diagnosis of kidneys with interstitial nephritis in pigs.(AU)
O propósito desse estudo foi avaliar as lesões histológicas observadas em rins condenados por nefrite pelo Serviço de Inspeção Federal, em dois frigoríficos de Mato Grosso, Brasil. Foram coletados 400 rins condenados por nefrite e submetidos aos exames de histologia, imuno-histoquímica (IHC) para Circovirus suíno Tipo 2 (PCV2), imunofluorescência direta (IF) para Leptospira sp. e reação em cadeia pela polimerase (PCR) para detecção de PCV2, Parvovirus suíno (PPV) e Torque teno vírus Tipo 1 e 2 (TTV1 e TTV2). Foram observadas lesões histológicas em 81% das amostras, sendo nefrite intersticial mononuclear a mais freqüente (77,50%). Das lesões de nefrite intersticial encontradas, 40,97% apresentaram padrão folicular. Através da PCR foi observada ampla distribuição dos agentes (PCV2, PPV, TTV1 e TTV2) nas propriedades e municípios, com ocorrência de 27,25%, 28,50%, 94% e 87,50%, respectivamente. Leptospira sp. não foi detectada através da IF. Houve associação significativa da infecção do PCV2 com presença de lesão histológica (P=0,008) e de células gigantes (P=0,0016). Também houve associação entre a co-infecção TTV2 e TTV1 (P<0,0001). Esses achados indicam que os vírus PCV2, PPV, TTV1 e TTV2 devem ser considerados no diagnóstico diferencial de rins com nefrite intersticial em suínos.(AU)
Assuntos
Animais , Doenças dos Suínos , Nefrite Intersticial/veterinária , Autopsia/veterinária , Rim/fisiopatologia , Circovirus/isolamento & purificação , Parvovirus Suíno/isolamento & purificação , Torque teno virus/isolamento & purificaçãoRESUMO
This study evaluated histological lesions in kidney samples from pigs with nephritis in two slaughterhouses in the State of Mato Grosso, Brazil. Four hundred samples were subjected to histology, anti-porcine circovirus type 2 (PCV2) immunohistochemistry (IHC), anti-Leptospira sp. immunofluorescence (IF), and polymerase chain reaction (PCR) for PCV2, porcine parvovirus (PPV), and Torque teno virus type 1 and 2 (TTV1, TTV2) detection. Histological lesions were found in 81% of the samples, and mononuclear interstitial nephritis was the most frequent lesion (77.50%). A follicular pattern was observed in 40.97% of the interstitial nephritis lesions. PCV2, PPV, TTV1, and TTV2 were identified in the kidneys by PCR in 27.25%, 28.50%, 94%, and 87.5% of the samples, respectively. Leptospira sp. was not detected through IF. Infection by PCV2 (PCR) and the presence of histological lesions (P=0.008) and giant cells (P=0.0016) were significantly associated. An association was observed between the TTV2-TTV1 co-infection (P<0.0001) and the risk for pathogenesis. These findings indicated that PCV2, PPV, TTV1, and TTV2 were widely distributed among pigs in the local farms and that the presence of these agents should be considered in the differential diagnosis of kidneys with interstitial nephritis in pigs.
O propósito desse estudo foi avaliar as lesões histológicas observadas em rins condenados por nefrite pelo Serviço de Inspeção Federal, em dois frigoríficos de Mato Grosso, Brasil. Foram coletados 400 rins condenados por nefrite e submetidos aos exames de histologia, imuno-histoquímica (IHC) para Circovirus suíno Tipo 2 (PCV2), imunofluorescência direta (IF) para Leptospira sp. e reação em cadeia pela polimerase (PCR) para detecção de PCV2, Parvovirus suíno (PPV) e Torque teno vírus Tipo 1 e 2 (TTV1 e TTV2). Foram observadas lesões histológicas em 81% das amostras, sendo nefrite intersticial mononuclear a mais freqüente (77,50%). Das lesões de nefrite intersticial encontradas, 40,97% apresentaram padrão folicular. Através da PCR foi observada ampla distribuição dos agentes (PCV2, PPV, TTV1 e TTV2) nas propriedades e municípios, com ocorrência de 27,25%, 28,50%, 94% e 87,50%, respectivamente. Leptospira sp. não foi detectada através da IF. Houve associação significativa da infecção do PCV2 com presença de lesão histológica (P=0,008) e de células gigantes (P=0,0016). Também houve associação entre a co-infecção TTV2 e TTV1 (P<0,0001). Esses achados indicam que os vírus PCV2, PPV, TTV1 e TTV2 devem ser considerados no diagnóstico diferencial de rins com nefrite intersticial em suínos.
Assuntos
Animais , Autopsia/veterinária , Nefrite Intersticial/veterinária , Rim/fisiopatologia , Doenças dos Suínos , Circovirus/isolamento & purificação , Parvovirus Suíno/isolamento & purificação , Torque teno virus/isolamento & purificaçãoRESUMO
This work aimed to detect and study natural co-infection of Circoviridae torque teno virus (TTV) and porcine circovirus 2 (PCV2) in the swine reproductive apparatus. Semen and organs from 17 boars were tested by nested and real-time PCR. PCV2 was amplified from semen (47%), lymph nodes (84.6%) and testicles (35.3%). TTV2 was amplified from 16/17 testis and 13/13 lymph nodes. TTV1 DNA was detected in fewer testicle samples (2/17), which were also TTV2 positive. Analyzed ovaries, follicular fluid and uteri of 83 culled sows showed TTV2, TTV1 and PCV2 from 49.3%, 30.1% and 6.0% of the sows, respectively. Sperm analysis indicated insignificant differences between PCV2 and TTVs positive and negative boars. The most frequent pathologic lesion in sows was endometritis (28.9%), but this was unassociated with PCV2 or TTVs detection. These findings question the importance of PCV2 and TTV2 natural co-infection in the pathology of porcine reproductive failures.
Assuntos
Circovirus/isolamento & purificação , Infecções por Vírus de DNA/veterinária , Doenças dos Genitais Femininos/veterinária , Doenças dos Genitais Masculinos/veterinária , Doenças dos Suínos/virologia , Torque teno virus/isolamento & purificação , Animais , Infecções por Vírus de DNA/virologia , Feminino , Doenças dos Genitais Femininos/virologia , Doenças dos Genitais Masculinos/virologia , Masculino , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Sêmen/virologia , SuínosRESUMO
Pseudorabies is a disease caused by pseudorabies virus (PRV) and is responsible for considerable economic losses in the swine industry. The objective of this work was to use molecular epidemiology as a tool to facilitate the study of PRV outbreaks in Brazil. The standard PRV strain Shope, the vaccine strain Bartha and isolates from the south and the southeast regions of Brazil, were amplified for gE and gC partial genes by PCR. Results indicated that Brazilian PRV isolates are grouped in two clusters, A and B, except for one isolate that grouped with Bartha and Shope. Most Brazilian PRV isolates belonged to cluster B and diverged from virus isolated from other countries.
Assuntos
Doenças dos Bovinos , Herpesvirus Suídeo 1/genética , Epidemiologia Molecular , Pseudorraiva/epidemiologia , Pseudorraiva/virologia , Doenças dos Suínos , Sequência de Aminoácidos , Animais , Sequência de Bases , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Herpesvirus Suídeo 1/classificação , Herpesvirus Suídeo 1/isolamento & purificação , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/virologia , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/genéticaRESUMO
Porcine circovirus infections are caused by the porcine circovirus 2 (PCV2). Among six different clinical manifestations involving respiratory, enteric, nervous and reproductive signs, the postweaning multisystemic wasting syndrome (PMWS) is the most important and studied disease. However, reproductive failures associated with PCV2 have been increasingly reported. Some studies have shown the possible contamination of sows by semen of PCV2 positive boars. In order to investigate the transmission of PCV2 by contaminated semen and its ability to infect the sow and piglets, 20 PCV2 negative sows were inseminated, 10 with negative boar semen and 10 with previously nested-PCR tested positive boar semen. The sows were weekly monitored and blood samples were collected. Based on the results, 4 out 20 sows were selected (1 sow was PCR negative and inseminated with a negative semen, 2 sows were PCR negative and inseminated with a positive semen and 1 sow was PCR negative and inseminated with a positive semen, but became PCR positive around the 30 days of pregnancy). After weaning, 12 male piglets, 3 of each sow, were selected and maintained under isolation. In order to investigate which organs harbored the virus, the young pigs were necropsied around 9 months of age. Samples of serum collected monthly were tested by immunocitochemistry (ICC), and all 12 pigs serum converted. Samples of lymphoid, systemic and reproductive organs were analyzed by nested-PCR and immunohistochemistry (IHC). Evaluation of the samples by nested-PCR, revealed that several tissues were positive in 10 of 12 pigs, mainly the lymph nodes, bone marrow and spleen. Various samples were positive by IHC in 8 of 12 piglets, being the lymph nodes, tonsils and bulbourethral glands the most frequently positive. Thus, the results of testing different samples, in the 3 tests (ICC, nested-PCR and IHC) were complementary. These results show that PCV2 transmission through semen to the sows...(AU)
Assuntos
Animais , Circovirus/isolamento & purificação , Infecções por Circoviridae/diagnóstico , Sêmen , Reação em Cadeia da Polimerase/métodos , SuínosRESUMO
Porcine circovirus infections are caused by the porcine circovirus 2 (PCV2). Among six different clinical manifestations involving respiratory, enteric, nervous and reproductive signs, the postweaning multisystemic wasting syndrome (PMWS) is the most important and studied disease. However, reproductive failures associated with PCV2 have been increasingly reported. Some studies have shown the possible contamination of sows by semen of PCV2 positive boars. In order to investigate the transmission of PCV2 by contaminated semen and its ability to infect the sow and piglets, 20 PCV2 negative sows were inseminated, 10 with negative boar semen and 10 with previously nested-PCR tested positive boar semen. The sows were weekly monitored and blood samples were collected. Based on the results, 4 out 20 sows were selected (1 sow was PCR negative and inseminated with a negative semen, 2 sows were PCR negative and inseminated with a positive semen and 1 sow was PCR negative and inseminated with a positive semen, but became PCR positive around the 30 days of pregnancy). After weaning, 12 male piglets, 3 of each sow, were selected and maintained under isolation. In order to investigate which organs harbored the virus, the young pigs were necropsied around 9 months of age. Samples of serum collected monthly were tested by immunocitochemistry (ICC), and all 12 pigs serum converted. Samples of lymphoid, systemic and reproductive organs were analyzed by nested-PCR and immunohistochemistry (IHC). Evaluation of the samples by nested-PCR, revealed that several tissues were positive in 10 of 12 pigs, mainly the lymph nodes, bone marrow and spleen. Various samples were positive by IHC in 8 of 12 piglets, being the lymph nodes, tonsils and bulbourethral glands the most frequently positive. Thus, the results of testing different samples, in the 3 tests (ICC, nested-PCR and IHC) were complementary. These results show that PCV2 transmission through semen to the sows...