Estabelecimento de escores histopatológicos de lesão hepática e determinação dos valores normais das enzimas aspartato aminotransferase e creatina quinase em frangos de corte

A aflatoxicose é uma doença causada pela ingestão de aflatoxinas, as quais constituem um grupo de metabólitos altamente tóxicos e carcinogênicos, produzidos principalmente pelos fungos Aspergillus flavus e Aspergillus parasiticus. O mecanismo de ação das micotoxinas na célula animal ocorre na maioria das vezes, através de alterações dos processos metabólicos básicos e de alterações da função mitocondrial, da síntese protéica e de ácidos nucléicos, sendo este último um dos principais sítios de ação das aflatoxinas. A aflatoxicose pode se apresentar de duas formas. A forma aguda se caracteriza por desordem hepática, hemorragias e alta mortalidade; e a crônica, pela queda na produção, penas arrepiadas, paralisia imunossupressão e diarréia. Foram estudados 18 lotes de frangos de corte com 28 dias de idade, provenientes de seis produtores de nível bom produtivo, seis de nível médio e seis produtores de baixo nível de produção; totalizando 180 aves. As aves foram pesadas e sacrificadas, sendo coletadas amostras de fígados e de soros. Os fígados foram fixados em formalina a 10%, para análise histopatológica e congelados a -20°C para teste de ELISA. Das amostras de soro foram realizados testes de dosagem enzimática de aspartato aminotransferase (AST) e creatina quinase (CK) na tentativa de confirmar a lesão hepática. Os cortes histológicos de fígado foram analisados e escores de lesão estabelecidos para necrose e vacuolização dos hepatócitos, hiperplasia dos ductos biliares e infiltração periportal de células inflamatórias. Os escores de cada lesão foram comparados com os níveis de aflatoxina obtidos pelo teste de ELISA, com o peso das aves e com os valores de AST e CK. Além disso, foram comparados os níveis de aflatoxina com o peso dos frangos e com os valores enzimáticos. Por fim, estes valores de AST e CK foram relacionados com o peso dos frangos. Foi possível concluir que, nas condições estudadas, os escores de lesões hepáticas são inversamente proporcionais aos níveis de aflatoxina detectados nos fígados de frangos com 28 dias de idade. Observou-se também, que não há relação entre os valores de AST, em função dos níveis de lesão hepática e dos valores de aflatoxina detectados; e a dosagem de AST não se presta como teste preliminar para detecção de lesão hepática causada pela ingestão de aflatoxina em frangos de corte aos 28 dias de idade

Classification of Avian Pathogenic Escherichia coli by a Novel Pathogenicity Index Based on an Animal Model

Background: Avian Pathogenic Escherichia coli is the main agent of colibacillosis, a systemic disease that causes considerable economic losses to the poultry industry. In vivo experiments are used to measure the ability of E. coli to be pathogenic. Generally, these experiments have proposed different criteria for results interpretation and did not take into account the death time. The aim of this study was to propose a new methodology for the classification of E. coli pathogenicity by the establishment of a pathogenicity index based in the lethality, death time and the ability of the strain to cause colibacillosis lesions in challenged animals.Materials, Methods & Results: A total of 293 isolates of E. coli were randomly selected to this study. The strains were isolated from cellulitis lesions, broiler bedding material or respiratory diseases and were previously confirmed through biochemical profile. The bacterial isolates were kept frozen at -20°C. The strains were retrieved from stocks and cultured in brain-heart infusion broth overnight at 37°C to obtain a final concentration of 109 UFC/mL. A total of 2940 one-dayold chicks from commercial breeding hens were randomly assigned to groups containing 10 animals and each group was subcutaneously inoculated in the abdominal region with 0.1 mL of the standard inoculum solution containing each of the strains. A control group of 10 broilers were inoculated with 0.1 mL of brain-heart infusion broth by the same route. The chicks were kept for seven days. They were observed at intervals of 6, 12 and 24 h post-inoculation during the first days. From the second day on, the chicks were observed at intervals of 12 h. According to the death time and to the scores of each lesion (aerosaculitis, pericarditis, perihepatitis, peritonitis and cellulitis), a formula to determine the Individual Pathogenicity Index was established. A value of 10 was established as the maximum pathogenicity rate for an inoculated bird. From this rate, 5 points corresponded to scores for gross lesions present at necropsy. For each lesion present, it represents 1 point. The remaining 5 points corresponded to the death time. To obtain the death time value, an index of 1, corresponding to the maximum value assigned to a death on the first day, was divided by the number of days that the birds were evaluated, resulting in a value of 0.1428, which corresponded to a survival bonus factor. It was possible to classify E. coli strains into four pathogenicity groups according to the pathogenicity index: high pathogenicity (pathogenicity index ranging from 7 to 10), intermediate pathogenicity (pathogenicity index ranging from 4 to 6.99), low pathogenicity (pathogenicity index ranging from 1 to 3.99) and apathogenic (pathogenicity index ranging from 0 to 0.99). The analysis of the strains according to their origin revealed that isolates from broiler bedding material presented a lower pathogenicity index.Discussion: It is possible that the source of isolation implies in different results, depending on the criteria adopted. This data reinforces the importance of use a more accurate mathematical model to represents the biological phenomenon. In the study, all avian pathogenic Escherichia coli strains were classified based on a pathogenicity index and the concept of the death time represents an interesting parameter to measure the ability of the strain to promote acute and septicemic manifestation. The use of a support method for poultry veterinary diagnostic accompanying the fluctuation of the bacteria pathogenicity inside the farms may indicate a rational use of antimicrobial in poultry industry

Flutuação estacional de protozoários e helmintos gastrintestinais na região de Eldorado do Sul, Rio Grande do Sul, Brasil : (resultados parciais)

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Intoxicação por ferro (hemocromatose) em um pato (Anas acuta) da região serrana do RS : relato de caso

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Characterization of immune and enteric systems of broilers after imunosupression with dexamethasone

Bursa of Fabricius (BF) and the thymus are primary lymphoid organs of poultry and play a major role in avian immunity. Enteric system is also involved in immunity. Several pathologic conditions directly impact BF and thymus size, and also affect intestinal parameters. Besides, there are several immune system depressor agents which affect birds. The selection of glucocorticoid as inducer of immunosuppression is applied in many experiments; however there are few studies that are applied to the reality in the field. In this context, the aim of this study was to evaluate the effects of dexamethasone as an inducer of immunosuppression on lymphoid organs and microscopic structures of the jejunum. Materials, Methods & Results: One-day-old chicks were used as a control group (n = 8) and the treated group (n = 25) received intramuscular dexamethasone on 21, 23, 24 and 26 day-old. Control birds and treated birds were euthanized 8, 16, 24, 32 and 40 h after inoculation; four control birds and six treated birds were euthanized on the eighth day after the last inoculation. Thymus, BF and jejunum were collected during the necropsy. The selected organs were processed, stained with hematoxylin and eosin and photographed. The BF and thymus cuts were evaluated by three histopathologists to determine the depletion score. Ten villi of each jejunum were evaluated for width and length of villi, depth crypt, microvillus length, enterocyte length of each villus, and wall thickness. Treated birds presented a mean weight lower than control group during all the experiment. The mean weight and the relative weight of the BF and thymus of control birds were significantly higher than treated ones. The lymphocyte depletion in BF and thymus scores differed significantly between groups, being higher in the group challenged with dexamethasone. There were no significant differences between groups for depth of crypt, height of core and height of microvilli. The intestines of the control group had higher mean values for length of villi, width of villi, height of enterocyte and thickness of wall. Significant correlation between weight of thymus and weight of BF, between weight of thymus and weight of bird and between weight of BF and bird weight were found. Discussion: Easy administration, low cost and the absence of suffering during inoculation make the use of glucocorticoids more advantageous to mimic immunosupression in poultry. Treatment with dexamethasone interfered directly in the weight of the birds. Evaluation of immune response of birds can be performed by the ratio of the BF weight in relation to the weight of the bird, as observed in the present study. The relative weight of the BF of untreated animals varied at all ages between 0.21% and 0.29%. It was also observed that the relative weight of BF and thymus in immunosuppressed birds tended to increase according to the end of the treatment. Depletion scores were higher in thymus, suggesting that glucocorticoid promoted more harmful effects on this organ. Dexamethasone had negative influence on length of villus, once mean value was 13% lower than those of control group. Width of villus and height of enterocyte core of treatment group were lower than those observed in control animals. Coefficient of determination greater than 70% was observed for almost all established relationships, except for the length of villi and weight of the thymus. Administration of dexamethasone promotes significant and negative effects on the gain of weight, length of villi, width of villi and height of enterocytes. These finding are useful for future experiments with controlled immunosuppression induction, once it provides significant information of the secondary effects of glucocorticoids administration in poultry

Characterization of Immune and Enteric Systems of Broilers after Imunosupression with Dexamethasone

Background: Bursa of Fabricius (BF) and the thymus are primary lymphoid organs of poultry and play a major role in avian immunity. Enteric system is also involved in immunity. Several pathologic conditions directly impact BF and thymus size, and also affect intestinal parameters. Besides, there are several immune system depressor agents which affect birds. The selection of glucocorticoid as inducer of immunosuppression is applied in many experiments; however there are few studies that are applied to the reality in the field. In this context, the aim of this study was to evaluate the effects of dexamethasone as an inducer of immunosuppression on lymphoid organs and microscopic structures of the jejunum.Materials, Methods & Results: One-day-old chicks were used as a control group (n = 8) and the treated group (n = 25) received intramuscular dexamethasone on 21, 23, 24 and 26 day-old. Control birds and treated birds were euthanized 8, 16, 24, 32 and 40 h after inoculation; four control birds and six treated birds were euthanized on the eighth day after the last inoculation. Thymus, BF and jejunum were collected during the necropsy. The selected organs were processed, stained with hematoxylin and eosin and photographed. The BF and thymus cuts were evaluated by three histopathologists to determine the depletion score. Ten villi of each jejunum were evaluated for width and length of villi, depth crypt, microvillus length, enterocyte length of each villus, and wall thickness. Treated birds presented a mean weight lower than control group during all the experiment. The mean weight and the relative weight of the BF and thymus of control birds were significantly higher than treated ones. The lymphocyte depletion in BF and thymus scores differed significantly between groups, being higher in the group challenged with dexamethasone. There were no significant differences between groups for depth of crypt, height of core and height of microvilli. The intestines of the control group had higher mean values for length of villi, width of villi, height of enterocyte and thickness of wall. Significant correlation between weight of thymus and weight of BF, between weight of thymus and weight of bird and between weight of BF and bird weight were found.Discussion: Easy administration, low cost and the absence of suffering during inoculation make the use of glucocorticoids more advantageous to mimic immunosupression in poultry. Treatment with dexamethasone interfered directly in the weight of the birds. Evaluation of immune response of birds can be performed by the ratio of the BF weight in relation to the weight of the bird, as observed in the present study. The relative weight of the BF of untreated animals varied at all ages between 0.21% and 0.29%. It was also observed that the relative weight of BF and thymus in immunosuppressed birds tended to increase according to the end of the treatment. Depletion scores were higher in thymus, suggesting that glucocorticoid promoted more harmful effects on this organ. Dexamethasone had negative influence on length of villus, once mean value was 13% lower than those of control group. Width of villus and height of enterocyte core of treatment group were lower than those observed in control animals. Coefficient of determination greater than 70% was observed for almost all established relationships, except for the length of villi and weight of the thymus. Administration of dexamethasone promotes significant and negative effects on the gain of weight, length of villi, width of villi and height of enterocytes. These finding are useful for future experiments with controlled immunosuppression induction, once it provides significant information of the secondary effects of glucocorticoids administration in poultry

Curva de eliminação de oocistos do gênero Eimeria spp. e identificação das espécies, em uma propriedade na região de Eldorado do Sul, Rio Grande do Sul, Brasil : resultados parciais

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Aflatoxicose em criação de marrecos no sul do Brasil

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Establishment of the histopathologics scores from hepatic lesion and determination of the normal values of enzymes aspartate aminotransferase and creatine kinase in chickens

A avicultura industrial apresenta altos índices produtivos, caracterizando-se pela alta tecnificação e pela eficiência. Inúmeros fatores podem prejudicar estes resultados, dentre os mais importantes citam-se as doenças imunodepressoras, as quais são agentes primários para o estabelecimento de inúmeros patógenos capazes de agravar o quadro clínico das aves, elevando ainda mais as perdas. Em vista disso, resolveu-se estudar um método mais eficaz para a determinação da depleção linfocitária da bolsa de Fabricius, órgão fundamental para a proliferação e maturação de linfócitos B. Dentre as doenças que mais acometem a bolsa de Fabricius citam-se a doença infecciosa da bolsa de Fabricius, as micotoxicoses e a anemia infecciosa. Foram utilizadas 50 amostras de bolsa de Fabricius coletadas intactas, processadas e o escore óptico de depleção estabelecido (de 1 a 5). As bolsas foram divididas em quadrantes e 12 folículos selecionados por amostra. As imagens foram adquiridas, analisadas com o software MATLAB® 6.5 e suas características extraídas. Com os dados foram geradas redes neurais (NEUROSHELL®), comparando-se os escores óticos e a classificação realizada pela rede. A rede foi capaz de classificar corretamente com alta sensibilidade (até 89,81%) e especificidade (até 96,17%) a maioria dos folículos, tendo um melhor desempenho utilizando-se três categorias (sensibilidade de até 79,39% e especificidade de até 91,94%) e duas categorias (sensibilidade e especificidade chegando a 92,54%). Os resultados mostraram que é possível a utilização de análise de imagem e redes neurais para a classificação histopatológica de depleção linfocitária da bolsa de Fabricius. A análise de imagem é uma ferramenta prática, com resultados objetivos, dimensiona o erro classificatório e padroniza a avaliação da depleção linfocitária bolsa.The industrial poultry has high productive indices, characterizing itself for the high technification and efficiency. Many factors can harm these results, the imunodepressives disease are amongst the most important causes. These diseases are the primary agents for the establishment of many secondary pathogens, aggravating the losses. In sight of this, we decided to study a more efficient method for the determination of the lymphoid depletion of bursa of Fabricius. The infectious bursal disease, the micotoxins and the infectious chicken anemia are the most important diseases of bursa of Fabricius. Fifty BF were examined by conventional optical microscopy and digital images were acquired and processed using MATLAB® 6.5 software. The ANN was generated using NEUROSHELL® CLASSIFIER software and the optical and digital data were compared. The ANN was able to make a comparable classification of digital and optical scores. The use of ANN was able to classify correctly the majority of the follicles with sensibility and specificity of 89% and 96%, respectively. When the follicles were scored and grouped in a binary fashion the sensibility and specificity increased significantly to 90 and 92%, respectively. These results demonstrate that the use of digital image analysis and ANN is a useful tool for the pathological classification of the BF lymphoid depletion. In addition

Estabelecimento de escores histopatológicos de lesão hepática e determinação dos valores normais das enzimas aspartato aminotrasferase e creatina quinase em frangos de corte.

A aflatoxicose é uma doença causada pela ingestão de aflatoxinas, as quais constituem um grupo de metabólitos altamente tóxicos e carcinogênicos, produzidos principalmente pelos fungos Aspergillus flavus e Aspergillus parasiticus. O mecanismo de ação das micotoxinas na célula animal ocorre na maioria das vezes, através de alterações dos processos metabólicos básicos e de alterações da função mitocondrial, da síntese protéica e de ácidos nucléicos, sendo este último um dos principais sítios de ação das aflatoxinas. A aflatoxicose pode se apresentar de duas formas. A forma aguda se caracteriza por desordem hepática, hemorragias e alta mortalidade; e a crônica, pela queda na produção, penas arrepiadas, paralisia imunossupressão e diarréia. Foram estudados 18 lotes de frangos de corte com 28 dias de idade, provenientes de seis produtores de nível bom produtivo, seis de nível médio e seis produtores de baixo nível de produção; totalizando 180 aves. As aves foram pesadas e sacrificadas, sendo coletadas amostras de fígados e de soros. Os fígados foram fixados em formalina a 10%, para análise histopatológica e congelados a -20°C para teste de ELISA. Das amostras de soro foram realizados testes de dosagem enzimática de aspartato aminotransferase (AST) e creatina quinase (CK) na tentativa de confirmar a lesão hepática. Os cortes histológicos de fígado foram analisados e escores de lesão estabelecidos para necrose e vacuolização dos hepatócitos, hiperplasia dos ductos biliares e infiltração periportal de células inflamatórias. Os escores de cada lesão foram comparados com os níveis de aflatoxina obtidos pelo teste de ELISA, com o peso das aves e com os valores de AST e CK. Além disso, foram comparados os níveis de aflatoxina com o peso dos frangos e com os valores enzimáticos. Por fim, estes valores de AST e CK foram relacionados com o peso dos frangos. Foi possível concluir que, nas condições estudadas, os escores de lesões hepáticas são inversamente proporcionais aos níveis de aflatoxina detectados nos fígados de frangos com 28 dias de idade. Observou-se também, que não há relação entre os valores de AST, em função dos níveis de lesão hepática e dos valores de aflatoxina detectados; e a dosagem de AST não se presta como teste preliminar para detecção de lesão hepática causada pela ingestão de aflatoxina em frangos de corte aos 28 dias de idade