Health assessment of raptors in triage in Belo Horizonte, MG, Brazil

Falconiformes (n=82), Strigiformes (n=84) and Cathartiformes (n=14) at a triage center (CETAS-Belo Horizonte, IBAMA, Brazil) were examined between 2008 and 2010 . No bird was reactive at hemagglutination-inhibition (HI) for antibodies against Mycoplasma gallisepticum (Mg). Two Caracara plancus (2/68) had HI titers (16-32) against Newcastle disease virus. No Chlamydophila psittaci DNA was detected in the liver (PCR; n=95). Blood smears (Giemsa; n=89) and spleen fragments (PCR; n=82) were 13.5% and 8.5% positive, respectively, for Haemoproteus only. Necropsy of Cathartiformes (n=10), Falconiformes (n=42) and Strigiformes (n=57) showed that trauma injuries were the main cause (63.3%) of admission and death, being fractures (38.5%) of the thoracic limbs (57.1%) the most frequent. Nematode (12.8%), cestode (1.8%), trematode (0.9%), and acanthocephalan (2.7%) parasite infections were relevant. Mites (Acari) were the most frequent (17.4%) external parasites, particularly Ornithonyssus sylviarum in Asio clamator and Amblyomma cajennense in Tyto alba. Chewing lice (10.1%) and Pseudolynchia spp. (9.2%) were also found. Histomonas spp. (6.4%) was found in the ceca of Bubo virginianus, Athene cunicularia, Tyto alba, and Asio clamator, but not in Falconiformes or Cathartiformes. Trichomonas spp. (oral cavity, pharynx and upper esophagus; 9.1%) was detected in Falconiformes and Strigiformes, but not in Cathartiformes. Trichomonas spp. were found in A. cunicularia, Asio clamator, Glaucidium brasilianum and Tyto alba (Strigiformes), and in Rupornis magnirostris, Milvago chimachima, Falco femoralis, Falco sparverius and Caracara plancus (Falconiformes). Coccidia (9.1%) (Sarcocystis spp., 6.4%) and mycosis were observed in most Tyto alba (70%). The evaluated Orders may not pose risks for commercial poultry production. Habitat loss and urban adaptation may be increasingly affecting raptors.

Detection of Mycoplasma gallisepticum in dead captive psittacines in Belo Horizonte, Brazil

Mycoplasma gallisepticum (Mg) infection of wild native Brazilian psittacines (Psittaciformes) which died of any cause during sorting, rehabilitation, or conservation, was investigated by PCR. Two previously described PCR methodologies using Mg specific primers were employed for the analyses of 140 swab samples (cloaca, trachea, or palatine cleft). Average positive Mg detection in cloacal swabs was 51.9%, with 80.0% (n=5) of Blue-and-yellow Macaws (Ara ararauna), 60.0% (n=3) Dusky Parrots (Pionus fuscus), 52.5% (n=59) Amazon Parrots (Amazona aestiva), 50.0% (n=2) Orange-winged Parrots (Amazona amazonica), 50.0% (n=2) Jandaya Parakeetsor Jandaya Conures (Aratinga jandaya), 0% (n=2) Golden Conures or Golden Parakeets (Guarouba guarouba), and 0% (n=2) Hyacinth Macaws (Anodorhynchus hyacinthinus). Palatine cleft swab sampling was more sensitive to detect Mg, with 85.4% (n=17) detection rate, as compared to 67.4% (n=46) obtained with tracheal samples, and 53.5% (n=77) with cloacal swabs. The surprisingly high Mg incidence in psittacines kept in conservation or triage environments is possibly due to the proximity or cohabitation with several bird species during confinement and housing psittacines of different origins together. The implementation of biosecurity measures and species-specific facilities is recommended.

Molecular studies of the Brazilian infectious bronchitis virus isolates

Avian infectious bronchitis virus (IBV) isolates have been widely characterized by reverse transcription followed by polymerase chain reaction and DNA sequencing. In present study, these techniques were applied to three viral genomic regions comprising the complete and/or a partial S1 segment, S2 and nucleocapsid genes. DNA sequences from viral isolates obtained from 1972 to 1989 and from 2006 to 2008 were compared. High similarity (>90%) was observed among some of the genomic segments, including S1 hypervariable region, which could suggest a common origin or ancestry. DNA sequences from S2 and N protein genes obtained from different infected tissues of the same flock were analyzed, and a clear segregation between respiratory and intestinal tract was observed. Therefore, these data suggest co-circulation of more than one viral strain in the same flock. 57.1% of DNA sequences from the S1 complete segment samples, 53.3% from the S2 fragment and 62.5% from the partial N gene were found to be different from analyzed sequences from reference strains leading to the conclusion that parte of viral isolates included in this study may be considered region specific. Considering the simultaneous analysis of the three genes, a large IBV genetic profile was observed in both old and recent isolates groups. However, most prominent diversity between viral isolates was obtained in the period from 1972 and 1989, showing the presence of a large number of variants in the state of Minas Gerais before the official approval of vaccination (1980).

Lead poisoning mortality in wild passeriformes and its detection in free-range chicken eggs in Southern Minas Gerais, Brazil

The mortality of passerines native of Southern Minas Gerais, Brazil, in August 2006 and the examination of sentinel free-range chicken eggs in August 2007 in an area surrounding a car battery lead smelter plant is described. The high levels of lead, as detected in the passerines (4.80-12.74 mg/kg) at the onset of mortality and at the follow-up sampling of the free-range chicken eggs (25.02-35.21 mg/kg in shells, 0.41-1.36 mg/kg in yolks and 0.40-0.75 mg/kg in the albumins), used as environmental sentinels, indicated the continuing lead pollution.

Lead poisoning mortality in wild passeriformes and its detection in free-range chicken eggs in Southern Minas Gerais, Brazil

The mortality of passerines native of Southern Minas Gerais, Brazil, in August 2006 and the examination of sentinel free-range chicken eggs in August 2007 in an area surrounding a car battery lead smelter plant is described. The high levels of lead, as detected in the passerines (4.80-12.74 mg/kg) at the onset of mortality and at the follow-up sampling of the free-range chicken eggs (25.02-35.21 mg/kg in shells, 0.41-1.36 mg/kg in yolks and 0.40-0.75 mg/kg in the albumins), used as environmental sentinels, indicated the continuing lead pollution.

Molecular studies of the Brazilian infectious bronchitis virus isolates

Avian infectious bronchitis virus (IBV) isolates have been widely characterized by reverse transcription followed by polymerase chain reaction and DNA sequencing. In present study, these techniques were applied to three viral genomic regions comprising the complete and/or a partial S1 segment, S2 and nucleocapsid genes. DNA sequences from viral isolates obtained from 1972 to 1989 and from 2006 to 2008 were compared. High similarity (>90%) was observed among some of the genomic segments, including S1 hypervariable region, which could suggest a common origin or ancestry. DNA sequences from S2 and N protein genes obtained from different infected tissues of the same flock were analyzed, and a clear segregation between respiratory and intestinal tract was observed. Therefore, these data suggest co-circulation of more than one viral strain in the same flock. 57.1% of DNA sequences from the S1 complete segment samples, 53.3% from the S2 fragment and 62.5% from the partial N gene were found to be different from analyzed sequences from reference strains leading to the conclusion that parte of viral isolates included in this study may be considered region specific. Considering the simultaneous analysis of the three genes, a large IBV genetic profile was observed in both old and recent isolates groups. However, most prominent diversity between viral isolates was obtained in the period from 1972 and 1989, showing the presence of a large number of variants in the state of Minas Gerais before the official approval of vaccination (1980).

Detection of Mycoplasma gallisepticum in dead captive psittacines in Belo Horizonte, Brazil

Mycoplasma gallisepticum (Mg) infection of wild native Brazilian psittacines (Psittaciformes) which died of any cause during sorting, rehabilitation, or conservation, was investigated by PCR. Two previously described PCR methodologies using Mg specific primers were employed for the analyses of 140 swab samples (cloaca, trachea, or palatine cleft). Average positive Mg detection in cloacal swabs was 51.9%, with 80.0% (n=5) of Blue-and-yellow Macaws (Ara ararauna), 60.0% (n=3) Dusky Parrots (Pionus fuscus), 52.5% (n=59) Amazon Parrots (Amazona aestiva), 50.0% (n=2) Orange-winged Parrots (Amazona amazonica), 50.0% (n=2) Jandaya Parakeetsor Jandaya Conures (Aratinga jandaya), 0% (n=2) Golden Conures or Golden Parakeets (Guarouba guarouba), and 0% (n=2) Hyacinth Macaws (Anodorhynchus hyacinthinus). Palatine cleft swab sampling was more sensitive to detect Mg, with 85.4% (n=17) detection rate, as compared to 67.4% (n=46) obtained with tracheal samples, and 53.5% (n=77) with cloacal swabs. The surprisingly high Mg incidence in psittacines kept in conservation or triage environments is possibly due to the proximity or cohabitation with several bird species during confinement and housing psittacines of different origins together. The implementation of biosecurity measures and species-specific facilities is recommended.

Occurrence of chicken anemia virus in backyard chickens of the metropolitan region of Belo Horizonte, Minas Gerais

The occurrence of CAV in backyard chickens in the metropolitan area of Belo Horizonte, Brazil, was evaluated. The spleen and thymus of chickens from different origins were collected for DNA extraction and nested-PCR. CAV genome was detected in 30% of the flocks (n=20) examined. CAV origin for backyard chickens is speculated, taking into consideration its widespread incidence in the chicken industry, the contamination of live vaccines with CAV prior to its eradication from SPF flocks, and the use of attenuated CAV vaccines.

Occurrence of chicken anemia virus in backyard chickens of the metropolitan region of Belo Horizonte, Minas Gerais

The occurrence of CAV in backyard chickens in the metropolitan area of Belo Horizonte, Brazil, was evaluated. The spleen and thymus of chickens from different origins were collected for DNA extraction and nested-PCR. CAV genome was detected in 30% of the flocks (n=20) examined. CAV origin for backyard chickens is speculated, taking into consideration its widespread incidence in the chicken industry, the contamination of live vaccines with CAV prior to its eradication from SPF flocks, and the use of attenuated CAV vaccines.

Estudo comparativo da virulência de amostras de vacina do vírus da doença de Newcastle em galinhas SPF por meio da análise morfométrica da espessura traqueal

Virulence of three vaccinal lentogenic strains (La Sota, Ulster and VG-GA) of the Newcastle disease virus (NDV) was determined by morphometric analysis of tracheal thickness, in 45-day-old SPF chickens (n=12), free from NDV antibodies. Tracheal thickness was evaluated 1, 2, 3, 4, 6, and 8 days after intratracheal vaccination. La Sota strain induced higher tracheal swelling than the others, during most of the time of the experiment. Maximum swelling of tracheal mucosa occurred at the thirdday after vaccination. At that time, La Sota and Ulster had the same virulence, and both caused higher swelling of tracheal mucosa than VG-GA strain.

A virulência das amostras de vacinas lentogênicas La Sota, Ulster e VG-GA do vírus da doença de Newcastle (VDN) foi determinada por análise morfométrica da espessura da traquéia de galinhas (n=12) de 45 dias de idade, livres de anticorpos anti-VDN, vacinadas por via intra-traqueal. As traquéias foram avaliadas 1, 2, 3, 4, 6 e 8 dias pós-vacinação. A amostra La Sota induziu maior espessamento da traquéia no decorrer de todo período experimental. Ao terceiro dia pós-vacinação, período em que as traquéias se apresentaram mais espessas, as amostras La Sota e Ulster não diferiram em sua virulência, sendo ambas mais virulentas do que a amostra VG-GA, induzindo maior espessamento de traquéia e causando lesões histológicas mais severas.

Production of monoclonal antibodies against conserved components of infectious bronchitis virus

Murine hybridomas producing IgG1 monoclonal antibodies (Mabs) against N and S2 proteins (53KDa and 82KDa, respectively) from avian infection bronchitis virus (IBV) strain M41 were generated by the fusion of a myeloma cell line (Sp2/0-Ag14) with spleen cells from Balb/c mice previously immunized with whole virus IBV M41. Post-fusion screening criterion was by ELISA and 36 positive hybrids were generated after fusions. Two hybrids specific to N (N3F10) and S2 (S12B2) proteins from M41 (serotype Massachusetts) were selected by western blotting. These Mabs recognized the Ark-99 (serotype Arkansas) and A5968 (serotype Connecticut) IBV strains in addition to M41. By ELISA, the Mab against the S2 (S12B2) recognized all reference and Brazilian strains (M41, SE-17, H52, 297, 283, PM-1, PM-2, PM-3, 351, 29-78 E 327) studied, while the Mab against N recognized only six (M41, SE-17, H52, 283, 327 e 297) strains. The Mab against S2 may become a useful tool for IBV detection on the routine diagnosis of infectious bronchitis, especially for helping the differential diagnosis of clinically and pathologically confusing diseases, while the Mab against N (N3F10) recognized a probably less conserved region among the strains and may be interesting to comparing IBV isolates.

Foram produzidos anticorpos monoclonais (AcM) da subclasse IgG1 contra as proteínas N (53KDa) e S2 (82KDa) do vírus da bronquite infecciosa das galinhas (VBIG) amostra M41. Os híbridos secretores originaram-se da fusão entre células de mielomas da linhagem Sp2/0-Ag14 e linfócitos B de camundongos Balb/c previamente imunizados com o vírus completo. O primeiro critério de seleção foi por ELISA, no qual 36 híbridos originados de duas fusões reagiram positivamente; destes, foram selecionados dois AcM que reagiram contra as proteínas N (N3D4) e S2 (S12B2) do VBIG da amostra M41 (sorotipo Massachusetts) no western blotting. Os mesmos AcM foram também capazes de reconhecer as estirpes Ark-99 (sorotipo Arkansas) e A5968 (sorotipo Connecticut) do VBIG no western blotting. No ELISA, o AcM contra S2 (S12b2) reconheceu 11 estirpes das 11 estudadas (M41, SE-17, H52, 297, 283, PM-1, PM-2, PM-3, 351, 29-78 E 327), enquanto o AcM contra a proteína N reconheceu apenas seis estirpes (M41, SE-17, H52, 283, 327 e 297). O anticorpo monoclonal contra S2 comportou-se como uma boa ferramenta de diagnóstico do VBIG, independentemente do sorotipo que pode ser aplicado em ensaios de rotina laboratorial, especialmente no diagnóstico das doenças que se confundem com a BIG nas galinhas, enquanto o AcM contra N (N3F10) demonstrou reconhecer uma região menos conservada entre as estirpes de VBIG e pode ser útil em estudos comparativos entre isolados de VBIG.

An outbreak of malaria by Haemoproteus columbae in pigeons

Relata-se um surto de malária por Haemoproteus columbae em pombos-correio (Columba livia), caracterizado por um quadro agudo com mortalidade de 3% ao dia em aves de aspecto saudável ou crônico com prostração e fraqueza. O exame clínico das aves doentes evidenciou sinusite e conjuntivite e à necropsia traqueíte hemorrágica, aerossaculite, esplenomegalia, nefromegalia e hepatomegalia com hemorragias. O díptero hematófago da família Hippoboscidae Pseudolynchia canariensis foi encontrado inserido entre as penas dos indivíduos. Esfregaços de sangue periférico e cardíaco e impressões de baço e fígado corados por Giemsa permitiram a visualização de inclusões intraeritrocitárias características de Haemoproteus columbae

Produção e caracterização de anticorpos monoclonais contra o vírus da bronquite infecciosa das galinhas

Monoclonal antibodies (Mab) specific to IBV Massachusetts M41 were produced aiming to the development of assays for IBV diagnosis and research, such as for rapidly detecting or typing IBV isolates. Mabs were detected by ELISA and characterized based on the isotype (ELISA) or IBV structural specificity (western blotting) respectively as IgA, IgG1, IgG2a, IgG2b or IgM or as specific to S1 (42), S2 (34, 43 and 71), M (7 and 22), N (15 and 50) and S whole molecule (3, 18, 52 and 57), the major IBV structural proteins

Um painel de anticorpos monoclonais (AcM) específicos contra o vírus da bronquite infecciosa das galinhas (VBIG) amostra Massachusetts M41 foi desenvolvido com a finalidade de possibilitar ferramentas para futuros estudos de amostras locais de VBIG. Os clones produtores de AcM foram detectados e caracterizados por ELISA, e para a determinação quanto à especificidade ao componente de VBIG foi utilizado western blotting (WB). Os híbridos produtivos foram clonados por diluição limitante, expandidos in vitro e mantidos em nitrogênio líquido. As bandas protéicas reconhecidas em WB pelos AcM apresentaram pesos moleculares que variaram de 180 a 30 kDa. Oito AcM reconheceram apenas um polipeptídeo e quatro ligaram-se ao polipeptídeo S (3, 18, 52 e 57) inteiro, com a banda de peso molecular aproximado de 180 kDa. Cinco AcM (5, 12, 41, 70 e 72) ligaram-se em mais de uma banda de VBIG. Outros sete AcM não se ligaram a nenhum polipeptídeo de VBIG. Entre os AcM que apresentam perspectivas de utilização em pesquisa e diagnóstico de VBIG incluem-se os produzidos pelo clone 42, dirigido contra S1, os produzidos pelos clones 34, 43 e 71, dirigidos contra S2, pelos clones 7 e 22, dirigidos contra M e pelos clones 15 e 50, cujos AcM reagiram contra a nucleoproteína N.

Mortality by filarial nematodes in passeriformes of species Oryzoborus maximiliani in Brazil

Descreve-se a ocorrência de filariose em passeriformes da espécie Oryzoborus maximiliani (bicudo) mantidos em cativeiro. Os sinais clínicos incluiram insuficiência respiratória e prostração, evoluindo para decúbito lateral e morte. Todos os indivíduos capturados adoeceram e morreram em poucos dias. As lesões mais significativas foram encontradas nos pulmões, que estavam acinzentados na região adjacente ao saco aéreo abdominal. Impressões do pulmão observadas ao microscópio em 100 aumentos permitiram a visualização de grande número de formas alongadas típicas de nematódeo. Considerando suas dimensões e os relatos da literatura consultada, especulou-se a possibilidade de filariose. As condições de estresse de captura e cativeiro podem ser determinantes do quadro agudo observado, o que permite sugerir esta suspeita em casos semelhantes. Considera-se importante, entretanto, a possibilidade de a manifestação clínica na forma crônica ou assintomática poder ser mais comum que a aguda.

Cryopreservation of an avian spirochete strain in liquid nitrogen

Soros de aves experimentalmente infectadas, contendo espiroquetas viáveis, foram submetidos a dois procedimentos antes da criopreservação: glicerol na diluição de 1/2 (v/v), designado como soro com glicerol a 50% (GS), e dimetilsulfóxido na proporção de 1/10 (v/v), designado como soro com DMSO a 10% (DS). Após 15 meses de estocagem em nitrogênio líquido, amostras dos tratamentos GS e DS foram descongeladas e suas infectividades foram testadas em frangos susceptíveis. Apesar de ambos os procedimentos terem mantidos a infectividade da bactéria, DMSO a 10% no soro de frango apresentou-se mais satisfatório como criopreservante.