Detección y caracterización del virus de la anemia infecciosa aviar en Colombia

RESUMEN Objetivo. identificar el virus de la anemia infecciosa aviar (chicken anemia virus, CAV) en granjas avícolas y aves de traspatio en Antioquia, Colombia. Materiales y métodos. Se tomaron muestras de sangre y plumas de gallinas ponedoras; en cada granja se eligieron tres aves de seis edades diferentes (1, 15, 30, 60, 90 y 120 días de edad). También se obtuvieron muestras de aves de traspatio ubicadas cerca de las granjas estudiadas. Se realizó ELISA y PCR para el análisis de las muestras. Resultados. Mediante PCR, el 84% de las aves resultaron positivas al CAV en sangre total y el 66% en muestras de plumas. El 60% de las aves de traspatio dieron positivo en sangre y el 40% en folículo de pluma. Mediante ELISA, el 22% de las aves de las granjas avícolas presentó títulos de anticuerpos altos y el 19% moderados. En las aves de traspatio, el 43% presentó títulos de anticuerpos altos y 29% moderados. Además, los resultados de la prueba de RFLP y la secuenciación mostraron que el virus circulante encontrado en este estudio era diferente del de la cepa vacunal Cux-1 utilizada en el país. Conclusiones. El CAV está presente en Colombia en aves comerciales como de traspatio. Según los hallazgos, un alto porcentaje de las aves dieron positivo para la detección viral, aunque el número de aves positivas por anticuerpos fue bajo. Se requiere determinar las características del virus circulante para explicar la respuesta de anticuerpos obtenida.

ABSTRACT Objective. identify the presence of chicken anemia virus (CAV) in poultry farms and backyard chickens from Antioquia, Colombia. Materials and Methods. Blood and feather samples were taken from laying chickens; in each farm, three birds of six different ages (1, 15, 30, 60, 90, and 120 days old) were chosen randomly. Backyard chicken samples were also obtained near the research farms. We used serology and molecular techniques to analyze the samples. Results. By PCR, the 84% of the birds were positive in whole blood and 66% were positive in feather samples. The 60% of backyard chickens tested were positive in blood and 40% in feather follicle. By serology, the 22% of the poultry farm birds presented high antibody titers and 19% moderate antibody titers. In the backyard chickens, 43% of them presented high antibody titers and 29% moderate antibody titers. In addition, results from the RFLP test and sequencing showed that the circulating virus found in this study was different from the Cux-1 vaccine strain used in Colombia. Conclusions. CAV is present in Colombia in both commercial and backyard chickens. According to the findings, a high percentage of the birds tested positive for viral detection, whereas the number of birds that tested positive for antibodies was low. Thus, the characteristics of the circulating virus need to be determined to explain the antibody response observed in this study.

Fowl Aviadenovirus E associated with hepatitis-hydropericardium syndrome in broiler breeders / Síndrome da hepatite-hidropericárdio por Aviadenovirus e em reprodutores de corte

ABSTRACT: Fowl aviadenovirus (FAdV) is an important pathogen in the global poultry industry and the etiology of inclusion body hepatitis-hydropericardium syndrome (HHS) in chickens. Since the 1990s, several outbreaks of HHS have occurred in poultry producing areas, including South America. The coinfection of FAdV and chicken anemia virus (CAV) may markedly impact the incidence of HHS. This study describes an outbreak of HHS in coinfection with CAV in industrial broiler breeders and characterizes the FAdV isolate. The three-week-old male broiler breeders had pale bone marrow, enlarged and yellowish liver, splenomegaly, and atrophied thymus; one chicken was also found with hydropericardium. Virus isolation was performed in SPF chicken embryos of liver and thymus. Tissues of the naturally infected chickens and the inoculated embryos were evaluated by PCR and histopathology. All affected chickens and inoculated embryos were positive for FAdV and CAV. The inoculated embryos had enlarged, greenish and hemorrhagic livers, and 30% died within 7 days of inoculation. Phylogenetic analysis of the FAdV isolate hexon gene partial sequence enabled grouping with E species. The E species has recently become a relevant species in several countries. The association of FAdV with CAV in breeders is of further concern due to both being capable of vertical transmission. Within the last decade, a worldwide upsurge of HHS in broiler breeders owing to failed biosecurity has occurred. In this episode, the failure on biosecurity may have enabled challenge with both FAdV and CAV, with pathological synergism. The CAV-impaired adaptive immunity may have benefited the FAdV infection.

RESUMO: Adenovírus aviário (FAdV) é um importante patógeno na indústria avícola global e a etiologia da síndrome da hepatite por corpúsculo de inclusão-hidropericárdio (SHH) em galinhas. Desde a década de 1990, vários surtos de SHH foram descritos em todas as áreas de produção de aves, incluindo na América do Sul, e a coinfecção entre FAdV e vírus da anemia das galinhas (CAV) pode ser agravante para todos os aspectos da SHH. Objetiva-se descrever um surto de SHH em matrizes de frangos corte, caracterizar a estirpe de FAdV envolvida e destacar a coinfecção com CAV. Foram avaliados machos reprodutores de corte com três semanas de idade, com medula óssea pálida, fígado aumentado e amarelado e esplenomegalia, timo atrofiado e um com hidropericárdio. Fígado e timo foram coletados para isolamento do vírus em embriões de galinhas SPF, PCR e histopatologia. Todas as aves acometidas e embriões inoculados foram positivos para FAdV e CAV. Os embriões inoculados tiveram óbito de 30% em até sete dias após a inoculação e alterações hepáticas por fígados esverdeados e aumentados. A análise filogenética de FAdV com base em parte da sequência do gene que codifica a proteína hexon revelou identidade com a espécie E, que se tornou disseminada em vários países. A coinfecção de FAdV e CAV resulta em maior intensidade de lesões, maior morbidade e mortalidade e em reprodutores tem alta relevância epidemiológica, em razão da transmissão vertical de ambos e da ampla distribuição geográfica das progênies infectadas. Na última década, ocorreu um aumento mundial na ocorrência de SHH em frangos de corte relacionado a falhas em biosseguridade, especialmente em reprodutores, condição que pode ter ocorrido neste episódio. A presença de FAdV e CAV em reprodutores é motivo para preocupação por reflexos negativos à imunidade e viabilidade das progênies.

Fowl Aviadenovirus E associated with hepatitis-hydropericardium syndrome in broiler breeders / Síndrome da hepatite-hidropericárdio por Aviadenovirus e em reprodutores de corte

Fowl aviadenovirus (FAdV) is an important pathogen in the global poultry industry and the etiology of inclusion body hepatitis-hydropericardium syndrome (HHS) in chickens. Since the 1990s, several outbreaks of HHS have occurred in poultry producing areas, including South America. The coinfection of FAdV and chicken anemia virus (CAV) may markedly impact the incidence of HHS. This study describes an outbreak of HHS in coinfection with CAV in industrial broiler breeders and characterizes the FAdV isolate. The three-week-old male broiler breeders had pale bone marrow, enlarged and yellowish liver, splenomegaly, and atrophied thymus; one chicken was also found with hydropericardium. Virus isolation was performed in SPF chicken embryos of liver and thymus. Tissues of the naturally infected chickens and the inoculated embryos were evaluated by PCR and histopathology. All affected chickens and inoculated embryos were positive for FAdV and CAV. The inoculated embryos had enlarged, greenish and hemorrhagic livers, and 30% died within 7 days of inoculation. Phylogenetic analysis of the FAdV isolate hexon gene partial sequence enabled grouping with E species. The E species has recently become a relevant species in several countries. The association of FAdV with CAV in breeders is of further concern due to both being capable of vertical transmission. Within the last decade, a worldwide upsurge of HHS in broiler breeders owing to failed biosecurity has occurred. In this episode, the failure on biosecurity may have enabled challenge with both FAdV and CAV, with pathological synergism. The CAV-impaired adaptive immunity may have benefited the FAdV infection.

Adenovírus aviário (FAdV) é um importante patógeno na indústria avícola global e a etiologia da síndrome da hepatite por corpúsculo de inclusão-hidropericárdio (SHH) em galinhas. Desde a década de 1990, vários surtos de SHH foram descritos em todas as áreas de produção de aves, incluindo na América do Sul, e a coinfecção entre FAdV e vírus da anemia das galinhas (CAV) pode ser agravante para todos os aspectos da SHH. Objetiva-se descrever um surto de SHH em matrizes de frangos corte, caracterizar a estirpe de FAdV envolvida e destacar a coinfecção com CAV. Foram avaliados machos reprodutores de corte com três semanas de idade, com medula óssea pálida, fígado aumentado e amarelado e esplenomegalia, timo atrofiado e um com hidropericárdio. Fígado e timo foram coletados para isolamento do vírus em embriões de galinhas SPF, PCR e histopatologia. Todas as aves acometidas e embriões inoculados foram positivos para FAdV e CAV. Os embriões inoculados tiveram óbito de 30% em até sete dias após a inoculação e alterações hepáticas por fígados esverdeados e aumentados. A análise filogenética de FAdV com base em parte da sequência do gene que codifica a proteína hexon revelou identidade com a espécie E, que se tornou disseminada em vários países. A coinfecção de FAdV e CAV resulta em maior intensidade de lesões, maior morbidade e mortalidade e em reprodutores tem alta relevância epidemiológica, em razão da transmissão vertical de ambos e da ampla distribuição geográfica das progênies infectadas. Na última década, ocorreu um aumento mundial na ocorrência de SHH em frangos de corte relacionado a falhas em biosseguridade, especialmente em reprodutores, condição que pode ter ocorrido neste episódio. A presença de FAdV e CAV em reprodutores é motivo para preocupação por reflexos negativos à imunidade e viabilidade das progênies.

Faecal Virome Analysis of Wild Animals from Brazil.

The Brazilian Cerrado fauna shows very wide diversity and can be a potential viral reservoir. Therefore, the animal's susceptibility to some virus can serve as early warning signs of potential human virus diseases. Moreover, the wild animal virome of this biome is unknown. Based on this scenario, high-throughput sequencing contributes a robust tool for the identification of known and unknown virus species in this environment. In the present study, faeces samples from cerrado birds (Psittacara leucophthalmus, Amazona aestiva, and Sicalis flaveola) and mammals (Didelphis albiventris, Sapajus libidinosus, and Galictis cuja) were collected at the Veterinary Hospital, University of Brasília. Viral nucleic acid was extracted, submitted to random amplification, and sequenced by Illumina HiSeq platform. The reads were de novo assembled, and the identities of the contigs were evaluated by Blastn and tblastx searches. Most viral contigs analyzed were closely related to bacteriophages. Novel archaeal viruses of the Smacoviridae family were detected. Moreover, sequences of members of Adenoviridae, Anelloviridae, Circoviridae, Caliciviridae, and Parvoviridae families were identified. Complete and nearly complete genomes of known anelloviruses, circoviruses, and parvoviruses were obtained, as well as putative novel species. We demonstrate that the metagenomics approach applied in this work was effective for identification of known and putative new viruses in faeces samples from Brazilian Cerrado fauna.

Development of real-time PCR assays for single and simultaneous detection of infectious bursal disease virus and chicken anemia virus.

Infectious bursal disease virus (IBDV) and chicken anemia virus (CAV) cause relevant immunosuppressive diseases in poultry. Clinical diagnosis of these viruses is challenging given the different disease presentations and the frequent occurrence of co-infections with other pathogens. Here, we standardized and validated simplex and duplex RT-qPCR assays for the straightforward detection of IBDV and CAV. The qPCR assays are based on primers and hydrolysis probes that target highly conserved regions of IBDV and CAV genomes. Analytical sensitivity tests on 10-fold serial dilutions containing 100-108 viral genomes indicated that the simplex assays have good determination coefficients and efficiency and detect a wide range of virus doses (102 to 108 molecules copies/reactions). The relatively small values of intra- and inter-assay variability ensure the repeatability and support its reproducibility in different diagnostic and research facilities. The assays are also efficient tools for absolute quantification as indicated by the analytical performance analysis. The assays have an excellent specificity and absence of cross-reactivity with negative samples, or with other common avian viruses. The simplex IBDV and CAV assays use probes labelled with different dyes (FAM and HEX) and can be multiplexed for the simultaneous detection of both viruses. The determination coefficients, PCR efficiencies, and relatively small intra- and inter-assay variability were comparable to the simplex assays. This duplex assay is the first to simultaneously detect IBDV and CAV using the same RNA extraction from the bursa of Fabricius in a single and straightforward step. Therefore, this method is time saving, provides quantitative results for both targets without any cross-reaction, and reduces the risk of carrying-over contaminations. The qPCR assays here developed can be used in simplex and duplex formats for detection and quantification of large number of samples with reliable sensitivity and specificity. These tools are expected to improve surveillance and control of these ubiquitous viruses.

Experimental coinfection of chicken anemia virus and Mycoplasma gallisepticum vaccine strains in broiler chicks

This study aimed at determining the clinical and pathological effects of the coinfection of young SPF chickens with chicken anemia virus (CAV) and Mycoplasma gallisepticum (MG) vaccine strains. The clinical signs, gross and microscopic lesions were determined after the experimental coinfection broilers with a CAV genotype 1 vaccine strain given intraperitoneally on the first day of age and a MG F-strain vaccine given intranasally on the 8th day of age. The experimental groups included the negative control (group 1), a group infected with the MG F-strain vaccine (group 2), and a group coinfected with CAV and MG vaccines (group 3). Chicks were examined clinically and post mortem at 23 days of age, and gross and microscopic lesions of the trachea, thymus, and air sacs were compared among treatments (Kruskal-Wallis test). Infections were confirmed by PCR for specific genetic fragments of each agent in the target tissues. Mortality was only observed in chicks on group 3, with two deaths and more severe lesions in the trachea, thymus and air sacs compared with groups 1 and 2 (p < 0.01). Dead chicks presented reduced thymus and spleen size, hemorrhagic trachea with catarrhal exudate and partial obstruction, pericarditis, catarrhal airsacculitis, lungs with liquid and ascites. The surviving chicks in group 3 showed more severe respiratory changes than those in group 2, in addition to thymus and spleen size reduction. Results indicate the adverse effects of the coinfection of young chickens with MG F-strain and CAV genotype 1 vaccines.

Experimental coinfection of chicken anemia virus and Mycoplasma gallisepticum vaccine strains in broiler chicks

This study aimed at determining the clinical and pathological effects of the coinfection of young SPF chickens with chicken anemia virus (CAV) and Mycoplasma gallisepticum (MG) vaccine strains. The clinical signs, gross and microscopic lesions were determined after the experimental coinfection broilers with a CAV genotype 1 vaccine strain given intraperitoneally on the first day of age and a MG F-strain vaccine given intranasally on the 8th day of age. The experimental groups included the negative control (group 1), a group infected with the MG F-strain vaccine (group 2), and a group coinfected with CAV and MG vaccines (group 3). Chicks were examined clinically and post mortem at 23 days of age, and gross and microscopic lesions of the trachea, thymus, and air sacs were compared among treatments (Kruskal-Wallis test). Infections were confirmed by PCR for specific genetic fragments of each agent in the target tissues. Mortality was only observed in chicks on group 3, with two deaths and more severe lesions in the trachea, thymus and air sacs compared with groups 1 and 2 (p < 0.01). Dead chicks presented reduced thymus and spleen size, hemorrhagic trachea with catarrhal exudate and partial obstruction, pericarditis, catarrhal airsacculitis, lungs with liquid and ascites. The surviving chicks in group 3 showed more severe respiratory changes than those in group 2, in addition to thymus and spleen size reduction. Results indicate the adverse effects of the coinfection of young chickens with MG F-strain and CAV genotype 1 vaccines.(AU)

The occurrence of orthoreovirus, rotavirus and chicken anemia virus in chickens of the poultry industry in Minas Gerais, Brazil / Ocorrência de orthoreovirus, rotavirus e vírus da anemia das galinhas em frangos de corte da indústria avícola de Minas Gerais

Fifty-four fecal samples taken from broiler chickens from 1 to 45 days of age, and of pullets from 10 to 13 weeks of age, original from eight different poultry regions in the state of Minas Gerais, Brazil, were collected from March 2008 to January 2010 for avian Orthoreovirus (ARV) and avian Rotavirus (AvRV) analyses. For the assay of ARV, RNA was immediately extracted (Trizolâ) and transcribed into cDNA for assaying in a nested-PCR with ARV-specific primers. For AvRV, polyacrylamide gel electrophoresis (PAGE) was performed with RNA extracts obtained by phenol-chloroform extraction. CAV was additionally investigated through a nested-PCR of thymus and spleen. Results found 5.55% positive for ARV and 9.25% for AvRV. Also, CAV and ARV genomes were detected in co-infection, in a highly prostrated and claudicating chicken flock. No ARV or AvRV infections were detected in pullets. Material of a clinically affected flock was inoculated into SPF embryos, resulting in embryonic hemorrhage, whitish foci in the chorio-allantoic membrane and death. Sequencing of ARV amplicons and isolate cDNA grouped local strains with the ARV S1133 strain, historically used in live vaccines, suggesting the continued circulation of this vaccine virus strain in intensive poultry regions. Detection rates for ARV and AvRV, as well as the presence of CAV, were additionally indicative of failing biosecurity strategies for the intensive poultry regions examined.

Avaliou-se a ocorrência de Orthoreovirus (ARV) e Rotavirus (AvRV) aviários na avicultura industrial de Minas Gerais. Foram colhidas cinquenta e quatro amostras de fezes de frangos de corte entre um e 45 dias e de frangas de postura de 10 a 13 semanas de idade. Para análise de ARV, o RNA foi imediatamente extraído (Trizol), transcrito em cDNA e avaliado em uma PCR com oligonucleotídeos iniciadores específicos para ARV. Para a investigação de AvRV, os extratos de RNA foram obtidos por fenol-clorofórmio e submetidos à eletroforese em gel de poliacrilamida. Todas as amostras foram também avaliadas para o DNA do vírus da anemia das galinhas (CAV) em uma nested-PCR específica. Em frangos de corte, a positividade encontrada para ARV foi de 5,55% e para AvRV de 9,25%. CAV foi detectado em coinfecção em um plantel com refugagem, claudicação e prostração. Nenhuma amostra de poedeiras foi positiva para ARV ou AvRV. Material de plantel com sinais clínicos foi purificado e inoculado em ovos SPF embrionados, sendo obtidas lesões hemorrágicas e focos brancos na membrana cório-alantóide. O sequenciamento dos produtos de PCR e de embrião agrupou os isolados de ARV com a estirpe S1133, historicamente usada como vacina viva. Os resultados sugerem a continuada circulação da infecção por estirpes assemelhadas a ARV S1133 nas regiões de avicultura industrial. Os índices de detecção de ARV, AvRV e CAV indicam que a intensificação nas regiões produtoras tem resultado em falhas de biosseguridade.

The occurrence of Orthoreovirus, Rotavirus and chicken anemia virus in chickens of the poultry industry in Minas Gerais, Brazil / Ocorrência de Orthoreovirus, Rotavirus e vírus da anemia das galinhas em frangos de corte da indústria avícola de Minas Gerais

Fifty-four fecal samples taken from broiler chickens from 1 to 45 days of age, and of pullets from 10 to 13 weeks of age, original from eight different poultry regions in the state of Minas Gerais, Brazil, were collected from March 2008 to January 2010 for avian Orthoreovirus (ARV) and avian Rotavirus (AvRV) analyses. For the assay of ARV, RNA was immediately extracted (Trizolâ) and transcribed into cDNA for assaying in a nested-PCR with ARV-specific primers. For AvRV, polyacrylamide gel electrophoresis (PAGE) was performed with RNA extracts obtained by phenol-chloroform extraction. CAV was additionally investigated through a nested-PCR of thymus and spleen. Results found 5.55% positive for ARV and 9.25% for AvRV. Also, CAV and ARV genomes were detected in co-infection, in a highly prostrated and claudicating chicken flock. No ARV or AvRV infections were detected in pullets. Material of a clinically affected flock was inoculated into SPF embryos, resulting in embryonic hemorrhage, whitish foci in the chorio-allantoic membrane and death. Sequencing of ARV amplicons and isolate cDNA grouped local strains with the ARV S1133 strain, historically used in live vaccines, suggesting the continued circulation of this vaccine virus strain in intensive poultry regions. Detection rates for ARV and AvRV, as well as the presence of CAV, were additionally indicative of failing biosecurity strategies for the intensive poultry regions examined.(AU)

Avaliou-se a ocorrência de Orthoreovirus (ARV) e Rotavirus (AvRV) aviários na avicultura industrial de Minas Gerais. Foram colhidas cinquenta e quatro amostras de fezes de frangos de corte entre um e 45 dias e de frangas de postura de 10 a 13 semanas de idade. Para análise de ARV, o RNA foi imediatamente extraído (Trizol), transcrito em cDNA e avaliado em uma PCR com oligonucleotídeos iniciadores específicos para ARV. Para a investigação de AvRV, os extratos de RNA foram obtidos por fenol-clorofórmio e submetidos à eletroforese em gel de poliacrilamida. Todas as amostras foram também avaliadas para o DNA do vírus da anemia das galinhas (CAV) em uma nested-PCR específica. Em frangos de corte, a positividade encontrada para ARV foi de 5,55% e para AvRV de 9,25%. CAV foi detectado em coinfecção em um plantel com refugagem, claudicação e prostração. Nenhuma amostra de poedeiras foi positiva para ARV ou AvRV. Material de plantel com sinais clínicos foi purificado e inoculado em ovos SPF embrionados, sendo obtidas lesões hemorrágicas e focos brancos na membrana cório-alantóide. O sequenciamento dos produtos de PCR e de embrião agrupou os isolados de ARV com a estirpe S1133, historicamente usada como vacina viva. Os resultados sugerem a continuada circulação da infecção por estirpes assemelhadas a ARV S1133 nas regiões de avicultura industrial. Os índices de detecção de ARV, AvRV e CAV indicam que a intensificação nas regiões produtoras tem resultado em falhas de biosseguridade.(AU)

Diverse uses of feathers with emphasis on diagnosis of avian viral infections and vaccine virus monitoring

The large amounts of feathers produced by the poultry industry, that is considered as a waste was explored for possible uses in various industries, such as meals for animals, biofuels, biodegradable plastic materials, combating water pollution and more. That review mentions these uses, but concentrate on the utilization of feathers for the diagnosis of viral infections and for monitoring vaccine viruses in chickens after vaccination. The viral diseases in which diagnosis using nucleic acids extracted from the feather shafts was described are, Marek's disease virus, circoviruses, chicken anemia virus, fowlpox virus, avian retroviruses, avian influenza virus and infectious laryngotracheitis virus. In two cases, of Marek's disease virus and of infectious laryngotracheitis virus, the differentiation of vaccine and wild-type viruses from feather shafts was made possible, thus allowing for monitoring the vaccination efficacy. The present review demonstrates also the stability of DNA viruses in feather shafts, and the possible evaluation of environmental dissemination of pathogens. When viruses are transmitted vertically, like in the cases of the retrovirus REV, a teratogenic effect on the development of feathers of the day-old newly hatched chick might occur in the case of avian influenza and the chicken anemia virus, which might indicate on a viral infection.

Diverse uses of feathers with emphasis on diagnosis of avian viral infections and vaccine virus monitoring

The large amounts of feathers produced by the poultry industry, that is considered as a waste was explored for possible uses in various industries, such as meals for animals, biofuels, biodegradable plastic materials, combating water pollution and more. That review mentions these uses, but concentrate on the utilization of feathers for the diagnosis of viral infections and for monitoring vaccine viruses in chickens after vaccination. The viral diseases in which diagnosis using nucleic acids extracted from the feather shafts was described are, Marek's disease virus, circoviruses, chicken anemia virus, fowlpox virus, avian retroviruses, avian influenza virus and infectious laryngotracheitis virus. In two cases, of Marek's disease virus and of infectious laryngotracheitis virus, the differentiation of vaccine and wild-type viruses from feather shafts was made possible, thus allowing for monitoring the vaccination efficacy. The present review demonstrates also the stability of DNA viruses in feather shafts, and the possible evaluation of environmental dissemination of pathogens. When viruses are transmitted vertically, like in the cases of the retrovirus REV, a teratogenic effect on the development of feathers of the day-old newly hatched chick might occur in the case of avian influenza and the chicken anemia virus, which might indicate on a viral infection.

A VP3/VP1 gene polymerase chain reaction assay for detection of chicken anemia virus in broiler samples / Teste de reação em cadeia de polimerase dos genes VP3 e VP1 para detecção do vírus da anemia das galinhas em amostras de campo

A PCR assay was designed for amplification of the highly conserved VP3 gene and a 5' region of the VP1 gene, for the diagnosis of CAV in organ samples of broiler flocks suspected of chicken infectious anemia. A comparison of the VP3/VP1 PCR with in vivo virus isolation revealed 100% agreement of the results, with 13 positive and 3 negative samples in both assays, indicating that the VP3/VP1 PCR is a specific diagnostic method. Tissues from additional 24 broiler chicken flocks, with CAV-like lesions and clinical history were then tested only by the VP3/VP1 PCR and a reference PCR with published primers for the VP1 gene. Nineteen samples resulted positive and one negative in both PCR, while another 4 samples were positive only in the VP3/VP1 PCR. These results indicate that the VP3/VP1 PCR is a sensitive, specific diagnostic test, suitable as an alternative to the expensive and time consuming in vivo virus isolation method, specially considering the difficult diagnosis of CAV strains not readily adaptable to MSB-1 cell culture.(AU)

Desenvolveu-se uma reação em cadeia de polimerase (PCR) para amplificação do altamente conservado gene VP3 e da região 5' do gene VP1, para o diagnóstico do vírus da anemia das galinhas (CAV), diretamente em amostras de campo de órgãos de frangos de corte com suspeita clínica da doença. A comparação entre o PCR VP3/VP1 com isolamento viral in vivo indicou 100% de concordância dos resultados, com 13 amostras positivas e três negativas em ambos os testes. Órgãos de outros 24 lotes de frangos com lesões e história clínica compatível com CAV foram testados com o PCR VP3/VP1 e com um PCR de referência com primers conhecidos para o gene VP1. Dezenove amostras resultaram positivas e uma negativa em ambos os PCR e quatro foram positivas apenas no PCR VP3/VP1. Estes resultados indicam que o PCR VP3/VP1 é um teste de diagnóstico sensível e específico, aplicável como alternativa ao método caro e demorado de isolamento viral in vivo e especialmente considerando-se amostras do CAV não adaptáveis a cultivos de células MSB-1.(AU)

Performance comparison between broilers positive and negative for antibodies against the chicken anemia virus

The chicken anemia virus (CAV) is present in virtually every country investigated, Brazil including. The aim of this study was to determine what the difference in performance is between positive (progeny of breeders vaccinated or with natural infection) and negative broilers to the presence of antibodies against the CAV in currently intensive raising systems. As a result, it was observed that negative broilers were significantly heavier than positive broilers. Negative males had a final weight 5.43% higher than positive males. There was no significant difference among different treatments in relation to parameters as mortality and feeding conversion. These study indicated that the presence of antibodies against CAV in broilers - may it be through vaccination or natural infection of breeders - did not generate progeny with superior performance under the tested raising conditions.

O vírus da anemia das galinhas (CAV - "chicken anemia virus") está presente em praticamente todos os países investigados, inclusive no Brasil. O objetivo deste trabalho foi determinar qual a diferença de desempenho, comparando frangos positivos (progênie de matrizes vacinadas ou com infecção natural) com frangos negativos para a presença de anticorpos contra o CAV, no sistema atual de criação intensiva. Como resultado, foi observado que os frangos negativos foram significativamente mais pesados que os frangos positivos. Os machos negativos tiveram um peso final 5,43% superior ao dos machos positivos. Não houve diferença significativa entre os tratamentos em relação aos parâmetros de mortalidade e conversão alimentar. Este estudo indicou que a presença de anticorpos contra o CAV em frangos de corte, seja através da vacinação ou infecção natural das matrizes, não gerou uma progênie com melhor desempenho nas condições de criação testadas.

Performance comparison between broilers positive and negative for antibodies against the chicken anemia virus

The chicken anemia virus (CAV) is present in virtually every country investigated, Brazil including. The aim of this study was to determine what the difference in performance is between positive (progeny of breeders vaccinated or with natural infection) and negative broilers to the presence of antibodies against the CAV in currently intensive raising systems. As a result, it was observed that negative broilers were significantly heavier than positive broilers. Negative males had a final weight 5.43% higher than positive males. There was no significant difference among different treatments in relation to parameters as mortality and feeding conversion. These study indicated that the presence of antibodies against CAV in broilers - may it be through vaccination or natural infection of breeders - did not generate progeny with superior performance under the tested raising conditions.

O vírus da anemia das galinhas (CAV - "chicken anemia virus") está presente em praticamente todos os países investigados, inclusive no Brasil. O objetivo deste trabalho foi determinar qual a diferença de desempenho, comparando frangos positivos (progênie de matrizes vacinadas ou com infecção natural) com frangos negativos para a presença de anticorpos contra o CAV, no sistema atual de criação intensiva. Como resultado, foi observado que os frangos negativos foram significativamente mais pesados que os frangos positivos. Os machos negativos tiveram um peso final 5,43% superior ao dos machos positivos. Não houve diferença significativa entre os tratamentos em relação aos parâmetros de mortalidade e conversão alimentar. Este estudo indicou que a presença de anticorpos contra o CAV em frangos de corte, seja através da vacinação ou infecção natural das matrizes, não gerou uma progênie com melhor desempenho nas condições de criação testadas.

Performance comparison between broilers positive and negative for antibodies against the chicken anemia virus

The chicken anemia virus (CAV) is present in virtually every country investigated, Brazil including. The aim of this study was to determine what the difference in performance is between positive (progeny of breeders vaccinated or with natural infection) and negative broilers to the presence of antibodies against the CAV in currently intensive raising systems. As a result, it was observed that negative broilers were significantly heavier than positive broilers. Negative males had a final weight 5.43% higher than positive males. There was no significant difference among different treatments in relation to parameters as mortality and feeding conversion. These study indicated that the presence of antibodies against CAV in broilers - may it be through vaccination or natural infection of breeders - did not generate progeny with superior performance under the tested raising conditions.

O vírus da anemia das galinhas (CAV - "chicken anemia virus") está presente em praticamente todos os países investigados, inclusive no Brasil. O objetivo deste trabalho foi determinar qual a diferença de desempenho, comparando frangos positivos (progênie de matrizes vacinadas ou com infecção natural) com frangos negativos para a presença de anticorpos contra o CAV, no sistema atual de criação intensiva. Como resultado, foi observado que os frangos negativos foram significativamente mais pesados que os frangos positivos. Os machos negativos tiveram um peso final 5,43% superior ao dos machos positivos. Não houve diferença significativa entre os tratamentos em relação aos parâmetros de mortalidade e conversão alimentar. Este estudo indicou que a presença de anticorpos contra o CAV em frangos de corte, seja através da vacinação ou infecção natural das matrizes, não gerou uma progênie com melhor desempenho nas condições de criação testadas.

Anticorpos Para o Vírus da Anemia das Galinhas (CAV) em Matrizes de Corte no Brasil

Although the isolation and identification of CAV as well as other recent diagnosis of the virus in broiler flocks have provided evidence for chicken anemia virus, Brentano & Ottati (1999), the prevalence of CAV had not yet been determined in Brazil. Since the knowledge of the prevalence of the virus and the level of antibodies against CAV in breeders flocks provide key information for designing control measures against chicken infectious anemia, we conducted a serological survey in broiler breeders of different ages, in five States with intensive commercial broiler flock production. Of a total of 2,355 sera tested by a commercial ELISA assay, 89% were found positive for antibodies to CAV, indicating a high prevalence of the virus in Brazil, in all the surveyed states. The analysis of antibodies to CAV in 6 to 70 week-old breeders showed that flocks between 6 and 18 weeks of age have the highest number of seronegative and low antibody titer chickens, thus representing the age group with the most uneven level of immunity against CAV. Chickens with negative or low antibody titers were still detected until 24 weeks of age, whereas at 25 weeks there was a 100% seroconversion to CAV and a reduction to less than 2,5% in the number of chickens with low antibody titers. These results provide evidence for a gradual and progressive virus spread in broiler breeder flocks, with the occurrence of a small percentage of chickens with low antibody titers at beginning of lay and therefore susceptible to inadequate transfer of passive immunity and vertical transmission of CAV to their progeny.

Apesar do diagnóstico da anemia infecciosa e mais evidências recentes de infecção em campo (Brentano & Ottati, 1999), não havia até hoje dados disponíveis da prevalência do vírus da anemia das galinhas (CAV) no Brasil. Uma vez que o conhecimento da prevalência do vírus e da distribuição de anticorpos nos lotes de matrizes é essencial para o delineamento de estratégias de controle da doença, realizamos um levantamento em 5 Estados com produção comercial intensiva de linhas de corte, abrangendo lotes de aves com diferentes idades. De 2.355 soros testados por ELISA, 89% foram positivos, indicando a alta prevalência do CAV no Brasil, em todos os Estados testados. A análise do soro de aves das 6 até as 70 semanas de idade demonstrou que lotes de 6 a 18 semanas apresentam desuniformidade na imunidade ao CAV, caracterizada por títulos de anticorpos abaixo dos níveis considerados protetores e aves ainda negativas para o CAV. A desuniformidade nos títulos de anticorpos e a ausência dos mesmos são evidentes ainda, mas em menor proporção também até as 24 semanas, enquanto que apenas partir das 25 semanas há 100% de soroconversão, acompanhada de significativa redução da desuniformidade no título de anticorpos das matrizes para menos de 2,5% das aves. Esses resultados fornecem evidência de uma progressiva disseminação do vírus em matrizes de corte, mas com riscos da presença de aves ainda susceptíveis à transmissão vertical e inadequada transferência de imunidade passiva no início do período de postura, que podem culminar em surtos da doença na progênie.

Anticorpos Para o Vírus da Anemia das Galinhas (CAV) em Matrizes de Corte no Brasil

Although the isolation and identification of CAV as well as other recent diagnosis of the virus in broiler flocks have provided evidence for chicken anemia virus, Brentano & Ottati (1999), the prevalence of CAV had not yet been determined in Brazil. Since the knowledge of the prevalence of the virus and the level of antibodies against CAV in breeders flocks provide key information for designing control measures against chicken infectious anemia, we conducted a serological survey in broiler breeders of different ages, in five States with intensive commercial broiler flock production. Of a total of 2,355 sera tested by a commercial ELISA assay, 89% were found positive for antibodies to CAV, indicating a high prevalence of the virus in Brazil, in all the surveyed states. The analysis of antibodies to CAV in 6 to 70 week-old breeders showed that flocks between 6 and 18 weeks of age have the highest number of seronegative and low antibody titer chickens, thus representing the age group with the most uneven level of immunity against CAV. Chickens with negative or low antibody titers were still detected until 24 weeks of age, whereas at 25 weeks there was a 100% seroconversion to CAV and a reduction to less than 2,5% in the number of chickens with low antibody titers. These results provide evidence for a gradual and progressive virus spread in broiler breeder flocks, with the occurrence of a small percentage of chickens with low antibody titers at beginning of lay and therefore susceptible to inadequate transfer of passive immunity and vertical transmission of CAV to their progeny.

Apesar do diagnóstico da anemia infecciosa e mais evidências recentes de infecção em campo (Brentano & Ottati, 1999), não havia até hoje dados disponíveis da prevalência do vírus da anemia das galinhas (CAV) no Brasil. Uma vez que o conhecimento da prevalência do vírus e da distribuição de anticorpos nos lotes de matrizes é essencial para o delineamento de estratégias de controle da doença, realizamos um levantamento em 5 Estados com produção comercial intensiva de linhas de corte, abrangendo lotes de aves com diferentes idades. De 2.355 soros testados por ELISA, 89% foram positivos, indicando a alta prevalência do CAV no Brasil, em todos os Estados testados. A análise do soro de aves das 6 até as 70 semanas de idade demonstrou que lotes de 6 a 18 semanas apresentam desuniformidade na imunidade ao CAV, caracterizada por títulos de anticorpos abaixo dos níveis considerados protetores e aves ainda negativas para o CAV. A desuniformidade nos títulos de anticorpos e a ausência dos mesmos são evidentes ainda, mas em menor proporção também até as 24 semanas, enquanto que apenas partir das 25 semanas há 100% de soroconversão, acompanhada de significativa redução da desuniformidade no título de anticorpos das matrizes para menos de 2,5% das aves. Esses resultados fornecem evidência de uma progressiva disseminação do vírus em matrizes de corte, mas com riscos da presença de aves ainda susceptíveis à transmissão vertical e inadequada transferência de imunidade passiva no início do período de postura, que podem culminar em surtos da doença na progênie.