RESUMO
The use of reference genes is required for relative quantification in gene expression analysis and the stability of these genes can be variable depending on the experimental design. Therefore, it is indispensable to test the reliability of endogenous genes previously to their use. This study evaluated nine candidate reference genes to select the most stable genes to be used as reference in gene expression studies with the femoral cartilage of normal and epiphysiolysis-affected broilers. The femur articular cartilage of 29 male broilers with 35 days of age was collected, frozen and further submitted to RNA extraction and quantitative PCR (qPCR) analysis. The candidate reference genes evaluated were GAPDH, HMBS, HPRT1, MRPS27, MRPS30, RPL30, RPL4, RPL5, and RPLP1. For the gene stability evaluation, three software were used: GeNorm, BestKeeper and NormFinder, and a global ranking was generated using the function RankAggreg. In this study, the RPLP1 and RPL5 were the most reliable endogenous genes being recommended for expression studies with femur cartilage in broilers with epiphysiolysis and possible other femur anomalies.
Assuntos
Doenças das Aves/genética , Cartilagem Articular/metabolismo , Galinhas/genética , Epifise Deslocada/veterinária , Algoritmos , Animais , Doenças das Aves/metabolismo , Galinhas/metabolismo , Epifise Deslocada/genética , Epifise Deslocada/metabolismo , Fêmur , Expressão Gênica , Perfilação da Expressão Gênica/estatística & dados numéricos , Perfilação da Expressão Gênica/veterinária , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Haemoproteid species have a wide global distribution, and they have been described in falcon species in several parts of the world. However, few studies in South America have focused on these birds. Haemoproteus spp. infections have been reported as the causative agents of serious histopathological changes, which can lead to the death of the host. Thus, this study aimed to molecularly and phylogenetically characterize Haemoproteus spp. in Caracara plancus, to characterize aspects of parasitism through clinical analysis and biochemical parameters, and to describe the histopathology of infection. To examine these aspects, 5 southern caracaras were examined clinically, and blood samples were collected. Blood smears were subsequently utilized in parasitemia calculations, PCR amplification, and serum biochemical investigations. Histological sections of the liver, kidneys, spleen, and heart were examined to check for possible histopathological changes. The birds showed clinical signs such as pallor and prostration that are consistent with Haemoproteus spp. infection. Moreover, the examination of the blood smears revealed 0.07% parasitemia in young gametocytes only. The PCR and sequencing results confirmed that the parasites belonged to Haemoproteus spp. The activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) enzymes, albumin, total serum proteins, and enzymatic urea were first described in C. plancus and serve as reference for future studies of bird species parasitized by Haemoproteus spp. Histopathology results showed signs of injury that were consistent with haemosporidian infection in the tissues of the analyzed organs. The present study is preliminary, and additional studies of Haemoproteus spp. infections in other bird species are needed to better understand the relationship between parasites and hosts, because despite the low parasitemia recorded, biochemical and histopathological changes in various organs were observed.
Assuntos
Doenças das Aves/parasitologia , Falconiformes/parasitologia , Haemosporida/genética , Infecções Protozoárias em Animais/parasitologia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Sequência de Bases , Doenças das Aves/metabolismo , Doenças das Aves/patologia , Proteínas Sanguíneas/análise , DNA de Protozoário/sangue , DNA de Protozoário/química , Haemosporida/classificação , Haemosporida/patogenicidade , Rim/patologia , Fígado/patologia , Miocárdio/patologia , Parasitemia/veterinária , Filogenia , Infecções Protozoárias em Animais/metabolismo , Infecções Protozoárias em Animais/patologia , Alinhamento de Sequência/veterinária , Baço/patologia , Ureia/sangueRESUMO
The aim of the present study was to evaluate the effects of heat stress (HS) and methionine supplementation on the markers of stress and on the gene expression levels of uncoupling proteins (UCP), betaine-homocysteine methyltransferase (BHMT), cystathionine ß-synthase (CBS), glutathione synthetase (GSS) and glutathione peroxidase 7 (GPx7). Broilers from 1 to 21 d and from 22 to 42 d of age were divided into three treatment groups related to methionine supplementation: without methionine supplementation (MD); recommended level of methionine supplementation (DL1); excess methionine supplementation (DL2). The broilers were either kept at a comfortable thermal temperature or exposed to HS (38°C for 24 h). During the starter period, we observed the effects of the interaction between diet and environment on the gene expression levels of UCP, BHMT and GSS. Higher gene expression levels of UCP and BHMT were observed in broilers that were maintained at thermal comfort conditions and received the MD diet. HS broilers fed the DL1 and DL2 diets had the highest expression level of GSS. The expression levels of the CBS and GPx7 genes were influenced by both the environment and methionine supplementation. During the grower period, the gene expression levels of BHMT, CBS, GSS and GPx7 were affected by the diet × environment interaction. A higher expression level of BHMT was observed in broilers maintained at thermal comfort conditions and on the MD diet. HS induced higher expression levels of CBS, GSS and GPx7 in broilers that received the DL1 and DL2 diets. The present results suggest that under HS conditions, methionine supplementation could mitigate the effects of stress, since methionine contributed to the increased expression levels of genes related to antioxidant activity.
Assuntos
Doenças das Aves/prevenção & controle , Dieta/veterinária , Regulação da Expressão Gênica no Desenvolvimento , Transtornos de Estresse por Calor/veterinária , Metionina/uso terapêutico , Estresse Oxidativo , Músculos Peitorais/enzimologia , Animais , Animais Endogâmicos , Antioxidantes/administração & dosagem , Antioxidantes/uso terapêutico , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Betaína-Homocisteína S-Metiltransferase/genética , Betaína-Homocisteína S-Metiltransferase/metabolismo , Biomarcadores/sangue , Biomarcadores/metabolismo , Doenças das Aves/dietoterapia , Doenças das Aves/metabolismo , Doenças das Aves/patologia , Galinhas , Ingestão de Energia , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Glutationa Sintase/genética , Glutationa Sintase/metabolismo , Transtornos de Estresse por Calor/dietoterapia , Transtornos de Estresse por Calor/metabolismo , Transtornos de Estresse por Calor/prevenção & controle , Homocisteína/sangue , Canais Iônicos/genética , Canais Iônicos/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Metionina/administração & dosagem , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Músculos Peitorais/metabolismo , Músculos Peitorais/patologia , Proteína Desacopladora 1 , Aumento de PesoRESUMO
An adult male Australian parakeet (Melopsittacus undulatus) presented a firm nodular lesion in the lateral metacarpal region of the right wing. Microscopically, there were neoplastic cells, round and polyhedral in shape, with abundant, slightly eosinophilic granular cytoplasm; they were strongly periodic-acid Schiff-positive and resistant to diastase digestion. Some groups of neoplastic cells were immunopositive for smooth muscle actin and desmin. There was no immunopositivity for S-100 protein, CD68 and cytokeratin. Ultrastructurally, the neoplastic cells were round and polygonal in shape, and they were characterized by abundant cytoplasm with numerous homogeneous osmophilic bodies covered by an electron-dense membrane (lysosomes). The histopathologic, immunohistochemical and ultrastructural features of the neoplastic tissue are consistent with a granular cell tumour, which has been described in different animal species and anatomic locations; however, this seems to be an infrequent neoplasm in Australian parakeets. The immunopositivity of the neoplastic cells for smooth muscle actin and desmin, as well as slight positivity for muscle with Masson's trichrome, suggest that this is a tumour of myogenic origin.
Assuntos
Doenças das Aves/patologia , Tumor de Células Granulares/veterinária , Melopsittacus , Neoplasias Cutâneas/veterinária , Animais , Austrália , Doenças das Aves/metabolismo , Desmina/metabolismo , Diagnóstico Diferencial , Tumor de Células Granulares/metabolismo , Tumor de Células Granulares/patologia , Tumor de Células Granulares/ultraestrutura , Imuno-Histoquímica/veterinária , Lisossomos/patologia , Lisossomos/ultraestrutura , Masculino , Microscopia Eletrônica/veterinária , Pele/patologia , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Neoplasias Cutâneas/ultraestruturaRESUMO
Poisoning of waterfowl due to ingestion of lead pellets is a worldwide problem in areas that are subject to hunting. No studies have assessed exposure of waterbirds to this heavy metal in Argentina, in spite of intense hunting activity, and the fact that only lead ammunition is commercially available. The objective of this study was to evaluate duck exposure to lead by examining gizzard and bone samples collected from 30 wild ducks, 16 Rosy-billed Pochard (Netta peposaca), and 14 Fulvous Whistling-Duck (Dendrocygna bicolor), provided by hunters in northern Santa Fe Province, Argentina, in July 2007. Radiographs, followed by dissection of the gizzards, showed that 31% of the Rosy-billed Pochards and 29% of the Fulvous Whistling-Ducks had ingested lead pellets (between one and four per animal). Lead in bone was found at concentrations associated with detrimental health effects. In spite of the small number of samples in this project, these results indicate high levels of lead exposure (both recent and chronic) in these species. This is the first report of a problem in Argentina that could represent a threat to the health and conservation of native aquatic species, their predators, and the wetlands they inhabit.