Survey for natural Neospora caninum infection in wild and captive birds

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

Microminerais complexados a aminoácidos no desempenho reprodutivo de matrizes pesadas e resposta da progênie

Este estudo teve como objetivo avaliar o uso de microminerais complexados a aminoácidos na alimentação de matrizes de frango de corte sobre o desempenho, variáveis de incubação e desempenho da progênie. O estudo foi dividido em três experimentos: exp.1. desempenho das matrizes, exp.2. incubação e exp.3. desempenho da progênie. No exp. 1, foram utilizadas 260 aves da linhagem CobbAvian 48 (240 matrizes e 20 machos), avaliando-se produção e massa de ovos, conversão alimentar por massa e por dúzia de ovos, peso e gravidade específica. No exp. 2, determinou-se a taxa de eclosão, eclodibilidade e fertilidade. No exp. 3 avaliou-se o desempenho dos pintinhos nascidos das incubações. No exp. 1, não houve interação entre tratamento e período (P>0,05) para produção de ovos, massa de ovos, conversão alimentar por massa e por dúzia de ovos, peso e gravidade específica dos ovos. Contudo, houve efeito (P<0,05) do período para todas as variáveis citadas anteriormente. No exp. 2, a taxa de eclosão, fertilidade e eclodibilidade não apresentaram diferenças significativas (P<0,05). Com relação ao exp. 3 também não houve diferença significativa (P<0,05) no desempenho da progênie. A suplementação com microminerais complexados a aminoácidos não influencia no desempenho das matrizes, variáveis de incubação e desempenho das progênies.

Sistema de alimentação de livre escolha com milho em grão ou moído e concentrado proteico para frangos de corte

O experimento teve como objetivo avaliar os sistemas de alimentação convencional ou de livre escolha, utilizando 6 dietas a base de milho em grão ou moído e concentrado protéico sobre o desempenho de frangos de corte do l° ao 45° dia de idade. O delineamento experimental foi inteiramente casualizado em esquema fatorial 6x2 (6 tratamentos e 2 sexos), com 3 repetições de 30 aves. Os tratamentos foram: T1-Milho moído + Concentrado do 1° ao 45° dia, fornecidos misturados; T2-Milho moído + Concentrado do 1° ao 45° dia, fornecidos separados em 2 comedouros; T3- Milho moído + Concentrado, fornecidos misturados, do l° ao 14° dia e do 15° ao 45° dia, milho em grão + concentrado, fornecidos misturados; T4 Milho moído + Concentrado, fornecidos misturados, do 1° ao 14° dia e do 15° ao 45° dia, milho em grão + concentrado, fornecidos separados; T5 - Milho moído + Concentrado, fornecidos misturados do 1° ao 21° dia e do 22° ao 45° dia, milho em grão + concentrado, fornecidos misturados; T6- Milho moído + Concentrado, fornecidos separados, do l ° ao 21° dia e do 22° ao 45° dia, milho em grão + concentrado, fornecidos misturados. Pêlos resultados e condições que realizou-se o presente trabalho conclui-se que: o milho em grão pode ser fornecido a frangos de corte misturado ao concentrado a partir do 15° dia de idade; o milho moído não proporcionou bons resultados de desempenho no programa de alimentação de livre escolha: as aves alimentadas com milho em grão apresentaram maior peso de moela, bem como acumularam mais gordura na carcaça; autilização de milho em grão fornecido junto ao concentrado, embora tenha proporcionado desempenho ligeiramente inferior à ração convencional, pode ser considerado satisfarório, sendo uma alternativa para pequenos produtores avícolas.

Trypanosoma cruzi: establishment of permeable cells for RNA processing analysis with drugs

Pre-mRNA maturation in trypanosomatids occurs through a process called trans-splicing which involves excision of introns and union of exons in two independent transcripts. For the first time, we present the standardization of Trypanosoma cruzi permeable cells (Y strain) as a model for trans-splicing study of mRNAs in trypanosomes, following by RNase protection reaction, which localizes the SL exon and intron. This trans-splicing reaction in vitro was also used to analyze the influence of NFOH-121, a nitrofurazone-derivative, on this mechanism. The results suggested that the prodrug affects the RNA processing in these parasites, but the trans-splicing reaction still occurred.

Rapid test for the evaluation of the activity of the prodrug hydroxymethylnitrofurazone in the processing of Trypanosoma cruzi messenger RNAs

No fully effective treatment has been developed since the discovery of Chagas' disease by Carlos Chagas in 1909. Since drug-resistant Trypanosoma cruzi strains are occurring and the current therapy is effectiveness in the acute phase but with various adverse side effects, more studies are needed to characterize the susceptibility of T. cruzi to new drugs. Many natural and/or synthetic substances showing trypanocidal activity have been used, even though they are not likely to be turned into clinically approved drugs. Originally, drug screening was performed using natural products, with only limited knowledge of the molecular mechanism involved in the development of diseases. Trans-splicing, which is unusual RNA processing reaction and occurs in nematodes and trypanosomes, implies the processing of polycistronic transcription units into individual mRNAs; a short transcript spliced leader (SL RNA) is trans-spliced to the acceptor pre-mRNA, giving origin to the mature mRNA. In the present study, permeable cells of T. cruzi epimastigote forms (Y, BOL and NCS strains) were treated to evaluate the interference of two drugs (hydroxymethylnitrofurazone - NFOH-121 and nitrofurazone) in the trans-splicing reaction using silver-stained PAGE analysis. Both drugs induced a significant reduction in RNA processing at concentrations from 5 to 12.5 µM. These data agreed with the biological findings, since the number of parasites decreased, especially with NFOH-121. This proposed methodology allows a rapid and cost-effective screening strategy for detecting drug interference in the trans-splicing mechanism of T. cruzi.

Comparação quantitativa da comunidade de aves de um fragmento de floresta semidecidual do interior do Estado de São Paulo em intervalo de 30 anos

Poucos estudos avaliaram em longo prazo a variação no tamanho populacional das espécies de aves em fragmentos florestais. Para avaliar a riqueza e a abundância específica da comunidade de aves de um remanescente de mata semidecidual do interior do Estado de São Paulo, sudeste do Brasil, foi conduzido o censo da avifauna florestal utilizando-se a metodologia de contagem em transecção. Estes resultados foram comparados com levantamento realizado na mesma localidade 30 anos antes, e as aves foram classificadas de acordo com suas categorias alimentares com a finalidade de associá-las à tendência ao aumento/diminuição de suas abundâncias após este intervalo de tempo. Embora tenha havido predominância de espécies com diminuição populacional, todas as categorias tróficas analisadas apresentaram também espécies com aumento em suas abundâncias. A maioria das espécies com propensão a deslocarem-se entre fragmentos apresentou diminuição em suas abundâncias. Sugerimos que, em relação a suas abundâncias específicas, as categorias tróficas são igualmente afetadas pelos processos da fragmentação, e que a regeneração florestal sofrida pelo remanescente pode ter resultado na perda de espécies de bordas. Espécies cujas abundâncias tenham reduzido neste intervalo de tempo podem sofrer extinção local futuramente.

Efeito da Proteína Bruta e de Diferentes Balanços Eletrolíticos das Dietas Sobre o Desempenho de Frangos no Período Inicial

Foram realizados dois experimentos utilizando-se 520 pintos machos de um dia para avaliar o efeito da proteína bruta (PB) e do balanço eletrolítico (Na + K- Cl) sobre o desempenho de frangos de corte no período inicial. O Experimento I (1 a 7 dias de idade das aves) foi realizado em baterias, utilizando-se 160 pintos machos Cobb, no Experimento II (1 a 21 dias) foram utilizados 360 pintos machos Aviam Farms que foram alojados em um galpão experimental dividido em box. Nos dois experimentos as aves foram aquecidas com lâmpadas infravermelhas e receberam água e ração à vontade. O delineamento foi inteiramente casualizado, em um esquema fatorial 2 x 2 (PB x balanço eletrolítico - BE), com 5 repetições e 8 aves por unidade experimental no Experimento I e fatorial 2 x 3 (PB x BE), com 3 repetições e 20 aves por unidade experimental no Experimento II. Os níveis de K foram mantidos constantes, oscilando-se o Na e o Cl para obter os balanços eletrolíticos desejados. Os parâmetro de desempenho (consumo de ração, ganho de peso e conversão alimentar) foram analisados no final de cada fase experimental. em ambos os experimentos não houve interação entre PB e BE. Os níveis de 21,0 e 23,5% de PB não afetaram o desempenho das aves. em dietas pré-iniciais e iniciais o melhor desempenho foi atingido com 260 mEq/kg.

Isolation of Salmonella enterica Serovar Enteritidis in Blue-Fronted Amazon Parrot (Amazona aestiva)

Avian salmonellosis is a disease caused by bacteria of the genus Salmonella that can cause three distinct diseases in birds: pullorum diseases, fowl typhoid, and paratyphoid infection. Various wildlife species are susceptible to infections by Salmonella, regardless of whether they live in captivity or freely in the wild. The present study verified the presence of Salmonella enterica serovar Enteritidis in three captive specimens of Amazona aestiva. The study involved a total of 103 birds undergoing rehabilitation to prepare for living in the wild, after having been captured from animal traffickers and delivered to the Centrofauna Project of the Floravida Institute in São Paulo, Brazil. This is the first report of Salmonella Enteritidis isolation in A. aestiva that originated from capture associated with animal trafficking; Salmonella was detected during the study by the serologic method of rapid serum agglutination on a plate with bacterial isolate. The antimicrobial profile exam of the isolated samples demonstrated sensitivity to ampicillin, cefaclor, ciprofloxacin, and cloranfenicol. The three samples also presented resistance to more than four antibiotics. The presence of the genes invA and spvC was verified by PCR technique and was associated with virulence and absence of class 1 integron, a gene related to antimicrobial resistance. The commercial antigen for pullorum disease was shown to be a useful tool for rapid detection in the screening of Salmonella of serogroup D(1) in Psittaciformes. New studies on Salmonella carriage in birds involved in trafficking must be performed to better understand their participation in the epidemiologic cycle of salmonellosis in humans and other animals.

Effects of adding polyclonal antibody preparations on ruminal fermentation patterns and digestibility of cows fed different energy sources

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

Two Surgical Approaches to Fracture Malunion Repair

Two birds were presented with malunion fractures. The first was a young toco toucan (Ramphastos toco) with malunion of the tarsometatarsus that was treated by an opening-corrective osteotomy and an acrylic-pin external skeletal fixator (type II) to stabilize the osteotomy. The second bird was m adult southern caracara (Caracara plancus) with radial and ulnar malunion that was treated by closing-wedge osteotomies. Stabilization of the osteotomy sites was accomplished through 1 bone plate fixed cranially on the ulna with 6 cortical screws and an interfragmentary single wire in radius. In both cases, the malunion was corrected, but the manus of the southern caracara was amputated because of carpal joint luxation that induced malposition of the feathers.

A double antibody sandwich ELISA for rapid diagnosis of virus infection and to measure the humoral response against infectious bursal disease on clinical material

A double antibody sandwich ELISA (DAS-ELISA) was developed and employed for simultaneous direct detection of infectious bursal disease virus (IBDV) from bursal samples and to measure the humoral response, using the same basic immunoreagents, the purified and non-purified antigen, capture antibody and chicken hyperimmune sera were prepared, and standardized for this purpose, the DAS-ELISA was applied to both 80 bursal suspensions and 224 corresponding serum samples from vaccinated and non-vaccinated commercial hocks, Bursae samples were collected at 2 weeks of age, and submitted to histological examination, virus isolation in specific pathogen-free chickens embryos, and the DAS-ELISA technique, Serum titres obtained in indirect ELISA and serum neutralization test were compared with those in DAS-ELISA, the agreement was 80% between DAS-ELISA, and the conventional techniques, with high sensitivity (87%) and specificity (90%).

Replication of classical infectious bursal disease virus in the chicken embryo related cell line

Infectious bursal disease (IBD) is an acute, highly contagious viral disease. The diagnosis of IBD depends on time-consuming and costly procedures, like virus isolation on chick embryos and histopathological examination, A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), immunoperoxidase and reverse transcription polymerase chain reaction (RT-PCR) were applied in this study to detect classical IBD virus (IBDV) after three blind passages of the Lukert strain on chicken embryo related (CER) cell monolayer after different periods of infection: 6, 12, 24 and 48 h, Cytophatic effects were most evident 12 h post-infection (p.i.) but were observed at 6 h p.i. The maximum discrimination between IBDV-infected and uninfected cell suspensions obtained by the use of DAS-ELISA for virus detection corresponded to 0.597+/-0.02 and 0.010+/-0.01 after 12h p.i., respectively. The RT-PCR was performed using the set of primers A3.1 and A3.2 to amplify the VP2 region of the IBDV genome, This molecular technique demonstrated that from 6 h p.i., it was possible to detect the viral RNA. The results show that the CER cell line can be used for classical IBDV propagation, confirmed by the DAS-ELISA, immunoperoxidase and RT-PCR assay.

Infectious bronchitis virus replication in the chicken embryo related cell line

The susceptibility of the chicken embryo related (CER) cell line to infectious bronchitis virus (IBV M41) was characterized after five consecutive passages in CER cells. Virus replication was monitored by cytopathic effect observation, electron microscopy, indirect immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR). At 96 h post-infection (p.i.), the cytopathic effect was graded 75% by cell fusion, rounding up of cells and monolayer detachment, and the electron microscopy image characterized by coronavirus morphology. Cytoplasmic fluorescence was readily observed by from 24 h p.i. onwards, and at all times the respective viral RNA from IBV-infected monolayers was demonstrated by RT-PCR. Extra-cellular virus was measured by virus titration performed on chicken kidney cells and embryonated chicken eggs, and respective titres ranged from 4.0 to 6.0 log(10) EID50/ml on embryonated chicken eggs, and from 2.0 to 6.0 log(10) TCID50/ml on both CER cells and chicken kidney cells studied from 24 to 120 h p.i. These results confirmed that the M41 strain replicated well in the CER cell line.

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.

Metabolismo oxidativo de heterófilos de avestruzes jovens

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