Detecção do TCoV (Turkey Coronavirus) a partir de amostras provenientes de perus (Meleagris gallopavo) com quadro agudo de enterite

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detection of turkey coronavirus in commercial turkey poults in Brazil

Poult enteritis complex has been incriminated as a major cause of loss among turkey poults in other countries. We have observed this in Brazil, associated with diarrhoea, loss of weight gain and, commonly, high mortality In this study, we have used the reverse transcriptase polymerase chain reaction (RT-PCR) to detect turkey coronavirus (TCoV) in sick poults 30 to 120 days of age from a particular producer region in Brazil. The RT-PCR was applied to extracts of intestine tissue suspensions, and the respective intestinal contents, bursa of Fabricius, faecal droppings and cloacal swabs. Primers were used to amplify the conserved 3' untranslated region of the genome, and the nucleocapsid protein gene of TCoV Histo pathological and direct immunohistochemical examinations were performed to detect TCoV antigen in infected intestine and bursa slides. All the results from stained tissues revealed lesions as described previously for TCoV infection. The direct immunohistochemical positive signal was present in all intestine slides. However, all bursa of Fabricius tissues analysed were negative. RT-PCR findings were positive for TCoV in all faecal droppings samples, and in 27% of cloacal swabs. Finally, the best field material for TCoV diagnosis was faecal droppings and/or intestine suspensions.

Pathology and Tissue Distribution of Turkey Coronavirus in Experimentally Infected Chicks and Turkey Poults

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Visual detection of turkey coronavirus RNA in tissues and feces by reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with hydroxynaphthol blue dye

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A new species of Tontelea from Amazonian Peru and Ecuador, and notes on the Tontelea attenuata species group (Celastraceae, Hippocrateoideae)

Tontelea fuliginea differs from other species whose stamens alternate with undivided stigma-lobes by its profusely ramified, many flowered, and densely dirty-brown puberulous inflorescences. This paper also provides a synopsis and key of the group of species with alternate stamens and 3-lobed stiomas with undivided lobes (T. attenuata group). Four lectotypes are designated for previously published names: Tontelea longifolia, Tontelea micrantha, Tontelea corrugulata, and Salacia micrantha var. lancifolia.

Isolation of bovine coronavirus (BCoV) in monolayers of HmLu-1 cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Epidemiological aspects of astrovirus and coronavirus in poults in the South Eastern Region of Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estabilidade de agregados de um latossolo vermelho tratado com cama de peru

Recentemente, tem ocorrido um grande aumento na produção de aves, com destaque para a carne de peru, motivado pelo aumento das exportações e a conquista de novos mercados. Como conseqüência, tem sido gerada uma grande quantidade de resíduos orgânicos com potencial impacto sobre o ambiente. A cama de peru constitui-se no principal resíduo dessa atividade avícola, e uma das formas de sua utilização é a sua disposição em solos como condicionador de suas características físicas e químicas. Assim, objetivou-se com este trabalho avaliar o efeito da aplicação da cama de peru na estabilidade de agregados de um Latossolo Vermelho Distrófico típico, sob pastagem de Brachiaria decumbens, localizado em Uberlândia, no Triângulo Mineiro, Minas Gerais. A cama de peru foi aplicada nas dosagens de: 0 (controle), 1.200, 2.400 e 4.800 kg ha-1 e 2.400 kg ha-1 + adubação mineral (36 kg ha-1 N, 60 kg ha-1 K2O e 60 kg ha-1de P2O5). As aplicações foram realizadas em janeiro de 2004 e o solo amostrado em duas épocas distintas (60 e 210 dias após aplicação) nas camadas de 0-20 e 20-40 cm para análise da estabilidade de agregados, avaliada pela percentagem total de agregados, percentagem de agregados maiores que 2mm, menores que 0,25mm e diâmetro médio geométrico (DMG). Não houve efeito das dosagens de cama de peru aplicadas na agregação do solo, entretanto, independentemente da dose aplicada houve um aumento na estabilidade de agregados, ao longo do tempo, notadamente na camada de 0-20 cm.

Fisheries and fishing effort at the Indigenous Reserves Ashaninka/Kaxinawá, river Breu, Brazil/Peru

Este artigo tem o objetivo de descrever a pesca de subsistência das populações tradicionais de uma aldeia Ashaninka e duas Kaxinawá vivendo à beira do rio Breu. Inicialmente, foram treinados monitores para preencher fichas de coleta de dados das pescarias nas aldeias durante um ciclo anual (agosto/1995 agosto/1996). A partir desses dados realizaram-se os inventários das espécies de peixes capturadas e dos ambientes pesqueiros. A análise dos dados foi efetuada por meio de estatística descritiva e exploratória. Os resultados obtidos foram os seguintes: i) os ambientes mais procurados pelos índios foram os poços; ii) as espécies mais capturadas os mandis (35%, Pimelodidae), os bodes ou cascudos (Loricariidae), com destaque para o bode praiano (25%, Hypostomus sp.), o curimatã (9%, Prochilodus sp.) e os saburus (8%, Curimatidae), entre outros; iii) constatou-se que os arreios ou apetrechos de pesca que mais capturam peixes são o tingui (veneno), a tarrafa e o arco/flecha, respectivamente; iv) durante o verão a atividade de pesca é mais intensa; v) as medidas de esforço de pesca e os fatores associados que foram estatisticamente significativos nas predições das capturas na Reserva Indígena foram: f1 = o (número de pescadores), f2 = o (número de pescadores*tempo total das pescarias) e f3 = o [(número de pescadores*tempo total das pescarias)-(o tempo de deslocamento)] e os fatores aldeias e arreios; vi) apesar da maioria das pescarias serem realizadas a pé até os pesqueiros, as capturas são maiores quando a locomoção se dá através de canoa a remo; e vii) os pescadores mais ativos nas pescarias na Reserva Indígena foram os Kaxinawá.

Validation of an immunohistochemistry assay to detect turkey coronavirus: A rapid and simple screening tool for limited resource settings

The objective of the present study was to develop and apply the direct immunohistochemistry (D-IHC) assay to search for turkey coronavirus (TCoV) antigens in formalin-fixed embedded-paraffin tissues by the use of biotin-labeled polyclonal antibody. Twenty-eight-day-old embryonated turkey eggs (n = 50) were inoculated with TCoV-purified virus, and 3 d after inoculation, sections from ileum, ileum-cecal junction, and ceca were harvested, fixed in neutral formalin, and embedded in paraffin blocks and used as positive control. In addition, a total of 100 field samples from ileum, ileum-cecal junction, and ceca, collected from 30 to 45-d-old turkeys poults experiencing an outbreak of acute enteritis, were used to search for TCoV by the same D-IHC. All results were compared with those obtained by conventional RT-PCR and indirect fluorescent antibody assay (IFA) for all tested samples. Turkey coronavirus was detected in experimentally infected embryo tissues and also in field samples in 100% of ileum-cecal junction and ceca by the 3 detection procedures. With IFA as a reference assay, sensitivity and specificity of D-IHC were 98 and 58%, whereas sensitivity and specificity of reverse transcription-PCR were 96 and 66%, calculated from the total of tested samples from experimental infection. Each of the examined procedures was highly specific (D-IHC, 93%; RT-PCR, 90%), sensitive (D-IHC, 85%; RT-PCR, 86%), and agreement of both D-IHC and RT-PCR was 99 and 100%, respectively, compared with IFA results obtained from all the field samples. These findings demonstrated the utility of D-IHC for direct detection of TCoV from field samples and considering the sensitivity and specificity found here, can be used as an alternative technique.

Myths and facts in the chronicles of the conquest of Ancient Peru

The chronicles of historical sources are difficult because they are a mix of myths and facts experienced by their authors. Starting from this premise, this study analyzes some of the Spanish and indigenous sources on the conquest of the Inca people and the description of this world and people who were so different. Among the chroniclers chosen, the article highlights the reports by Jose de Acosta on the Mundus Novus and by Cieza de Leon and Garcilaso de la Vega on the myths of the first Inca chief; finally, Francisco de Xerez, Juan de Betanzos, among others, on the event of the imprisonment and murder of Atahualpa, which resulted in the conquest of the "Tahuantinsuyu" by the Spanish.

Immunohistochemistry assay to detect Turkey coronavirus (TCoV) from experimentally infected poults

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Poult intestinal organ culture for propagation of Turkey coronavirus and assessment of host-virus interactions

Infection of young poults with turkey coronavirus (TCoV) produces a syndrome characterized by acute enteritis, diarrhea, anorexia, ruffled feathers, decreased body weight gain and uneven flock growth. The objective of this study was to standardize an intestinal organ culture (IOC) in order to assess host-virus interaction related to apoptosis. For this purpose the Brazilian strain (TCoV/Brazil/2006 with GenBank accession number FJ188401), was used for infection. Infected IOC cells had mitochondrial dysfunction and initial nuclear activation with MTT value of 90.7 (± 2.4) and apoptotic factor 2.21 (± 2.1), considered statistically different from uninfected IOC cells (p > 0.05). The kinetics of TCoV antigens and viral RNA was directly correlated to annexin-V, caspases- 2 and -3, p53, BCl-2 antigens at 24, 72 and 96 h post-infection (p.i.). Morphological and biochemical features of apoptosis, such as in situ nuclear fragmentation (TUNEL and annexin-V) and DNA ladder formation were also detected in infected cells at all assayed p.i. intervals. Moreover, different from other coronaviruses, the expression of both effective caspase-2 and - 3 and p53 antigens were considered lower. However, at all p.i., the BCl-2 antigens were expressed quantitatively and qualitatively as viral antigen measured by immunofluorescence microscopy analysis. Because the diagnosis of TCoV infection is only performed by infecting embryonated poult eggs, the pathological characteri tics related to host-virus interaction remain unclear. This is the first report on apoptosis of TCoV infected IOC, and reveals that it may be useful immunological method to assess virus pathogenesis.

Pathological and molecular analysis of Turkey coronavirus replication in captive wild Turkey embryos (broad-breasted bronze breeder)

The degree of genetic and pathologic variation exhibited by a turkey Coronavirus (TCoV) strain was investigated after nine serial passages in 25-day-old turkey embryos obtained from wild broad-breasted bronze breeders. In spite of spleen, liver, kidneys, cloacal bursa and thymus have been collected and analysed, the main histopathological changes were only documented in the intestine sections. Microscopic lesions were characterized as mild enteritis, low degree of enterocyte vacuolization and detachment of the intestinal villous after five consecutive passages and were considered absent in the last passages. Genealogic analysis based on S1 and S2 DNA sequences suggested that Brazilian isolate might be considered as originated from TCoV strains circulating in the United States, as 100% identity with TCoV-Gl strain. Although S1 S2 sequences from each passage revealed no significant point mutations, and no correlation could be speculate between S2 nucleotide changes and pathologic features in infected embryos. This is the first demonstration of wild turkey embryos as a model for TCoV isolation and propagation.