Classical scrapie diagnosis in ARR/ARR sheep in Brazil

Background: Scrapie is a transmissible spongiform encephalopathy (TSE) that affects sheep flocks and goat herds. The transfer of animals or groups of these between sheep farms is associated with increased numbers of infected animals and with the susceptibility or the resistance to natural or classical scrapie form. Although several aspects linked to the etiology of the natural form of this infection remain unclarified, the role of an important genetic control in scrapie incidence has been proposed. Polymorphisms of the PrP gene (prion protein, or simply prion), mainly in codons 136, 154, and 171, have been associated with the risk of scrapie.Case: One animal from a group of 292 sheep was diagnosed positive for scrapie in the municipality of Valparaíso, state of São Paulo, Brazil. The group was part of a flock of 811 free-range, mixed-breed Suffolk sheep of the two genders and ages between 2 and 7 years from different Brazilian regions. Blood was collected for genotyping (for codons 136, 141, 154 and 171), and the third lid and rectal mucosa were sampled for immunohistochemistry (IHC) for scrapie, from all 292 animals of the group. IHC revealed that seven (2.4%) animals were positive for the disease. Collection of samples was repeated for 90 animals, among which the seven individuals diagnosed positive and 83 other animals that had some degree of kinship with those. These 90 she...(AU)

Classical scrapie diagnosis in ARR/ARR sheep in Brazil

Background: Scrapie is a transmissible spongiform encephalopathy (TSE) that affects sheep flocks and goat herds. The transfer of animals or groups of these between sheep farms is associated with increased numbers of infected animals and with the susceptibility or the resistance to natural or classical scrapie form. Although several aspects linked to the etiology of the natural form of this infection remain unclarified, the role of an important genetic control in scrapie incidence has been proposed. Polymorphisms of the PrP gene (prion protein, or simply prion), mainly in codons 136, 154, and 171, have been associated with the risk of scrapie.Case: One animal from a group of 292 sheep was diagnosed positive for scrapie in the municipality of Valparaíso, state of São Paulo, Brazil. The group was part of a flock of 811 free-range, mixed-breed Suffolk sheep of the two genders and ages between 2 and 7 years from different Brazilian regions. Blood was collected for genotyping (for codons 136, 141, 154 and 171), and the third lid and rectal mucosa were sampled for immunohistochemistry (IHC) for scrapie, from all 292 animals of the group. IHC revealed that seven (2.4%) animals were positive for the disease. Collection of samples was repeated for 90 animals, among which the seven individuals diagnosed positive and 83 other animals that had some degree of kinship with those. These 90 she...

Development and application of an ERIC-PCR method for genotyping and differentiating Actinobacillus pleuropneumoniae isolates

Background: The pleuropneumonia caused by Actinobacillus pleuropneumoniae is one the most important swine respiratory diseases. Biochemical and serological tests are widely applied for the diagnosis and characterization of this bacterium. However, in some isolates, confl icting results are found. There are at least 15 serotypes with signifi cant differencesin virulence that have been identifi ed until now. Moreover, cross reactions between serotypes are not uncommon. Theserotype determination from isolates occurring in outbreaks is an important procedure in prophylaxis and control of thedisease. The present work focuses on the application of an ERIC-PCR technique for genotyping and differentiation ofA. pleuropneumoniae isolates.Materials, Methods & Results: Fifteen reference strains for the recognized A. pleuropneumoniae serotypes were analyzedin this work, alongside with 27 fi eld isolates that had been previously characterized regarding biochemical, serological andmolecular features. Total DNA from each sample was purifi ed and subjected to PCR amplifi cation using ERIC-specifi cprimers (ERIC1R and ERIC2). The resulting amplicons were analyzed by agarose gel electrophoresis and their sizes wereestimated from the gel images. Bands with similar sizes were identifi ed and used to construct a binary matrix that tookinto account the presence or absence of individual bands in all lanes. Pair-wise similarity coeffi cients were computed fromthe binary matrix and the similarity matrices obtained were utilized to construct an UPGMA-based dendrogram. The amplicons obtained from the A. pleuropneumoniae reference strains generated a very distinctive pattern for each one of thetested strains. Moreover, all samples presented a large enough number of amplicons (bands) as to enable an unequivocaldifferentiation of each sample. Reproducibility of the developed ERIC-PCR method was assessed by means of duplicate...(AU)

Development and application of an ERIC-PCR method for genotyping and differentiating Actinobacillus pleuropneumoniae isolates

Background: The pleuropneumonia caused by Actinobacillus pleuropneumoniae is one the most important swine respiratory diseases. Biochemical and serological tests are widely applied for the diagnosis and characterization of this bacterium. However, in some isolates, confl icting results are found. There are at least 15 serotypes with signifi cant differencesin virulence that have been identifi ed until now. Moreover, cross reactions between serotypes are not uncommon. Theserotype determination from isolates occurring in outbreaks is an important procedure in prophylaxis and control of thedisease. The present work focuses on the application of an ERIC-PCR technique for genotyping and differentiation ofA. pleuropneumoniae isolates.Materials, Methods & Results: Fifteen reference strains for the recognized A. pleuropneumoniae serotypes were analyzedin this work, alongside with 27 fi eld isolates that had been previously characterized regarding biochemical, serological andmolecular features. Total DNA from each sample was purifi ed and subjected to PCR amplifi cation using ERIC-specifi cprimers (ERIC1R and ERIC2). The resulting amplicons were analyzed by agarose gel electrophoresis and their sizes wereestimated from the gel images. Bands with similar sizes were identifi ed and used to construct a binary matrix that tookinto account the presence or absence of individual bands in all lanes. Pair-wise similarity coeffi cients were computed fromthe binary matrix and the similarity matrices obtained were utilized to construct an UPGMA-based dendrogram. The amplicons obtained from the A. pleuropneumoniae reference strains generated a very distinctive pattern for each one of thetested strains. Moreover, all samples presented a large enough number of amplicons (bands) as to enable an unequivocaldifferentiation of each sample. Reproducibility of the developed ERIC-PCR method was assessed by means of duplicate...

Polimorfismos de nucleotídeos únicos em 15 códons do gene da proteína priônica em um rebanho Suffolk afetado com scrapie no Brasil / Single nucleotide polymorphisms at 15 codons of the prion protein gene from a scrapie-affected herd of Suffolk sheep in Brazil

Scrapie is a transmissible spongiform encephalopathy of sheeps and goats, associated with the deposition of a isoform of the prion protein (PrPsc). This isoform presents an altered conformation that leads to aggregation in the host's central nervous and lymphoreticular systems. Predisposition to the prion agent infection can be influenced by specific genotypes related to mutations in amino acids of the PrPsc gene. The most characterized mutations occur at codons 136, 154 and 171, with genotypes VRQ being the most susceptible and ARR the most resistant. In this study we have analyzed polymorphisms in 15 different codons of the PrPsc gene in sheeps from a Suffolk herd from Brazil affected by an outbreak of classical scrapie. Amplicons from the PrPsc gene, encompassing the most relevant altered codons in the protein, were sequenced in order to determine each animal's genotype. We have found polymorphisms at 3 of the 15 analyzed codons (136, 143 and 171). The most variable codon was 171, where all described alleles were identified. A rare polymorphism was found at the 143 codon in 4 percent of the samples analyzed, which has been described as increasing scrapie resistance in otherwise susceptible animals. No other polymorphisms were detected in the remaining 12 analyzed codons, all of them corresponding to the wild-type prion protein. Regarding the risk degree of developing scrapie, most of the animals (96 percent) had genotypes corresponding to risk groups 1 to 3 (very low to moderate), with only 4 percent in the higher risks group. Our data is discussed in relation to preventive measures involving genotyping and positive selection to control the disease.(AU)

Scrapie é uma encefalopatia espongiforme transmissível de ovinos e caprinos, associado a deposição da isoforma da proteína priônica (PrPsc). Essa isoforma apresenta uma alteração conformacional que leva ao acúmulo da proteína no sistema nervoso central e linforeticular do hospedeiro. A predisposição a infecção pelo agente priônico pode ser influenciado por genótipos específicos relacionados a mutações na sequência de aminoácidos do gene PrPsc. As principais mutações caracterizadas ocorrem nos códons 136, 154 e 171, sendo o genótipo VRQ o mais suscetível e o genótipo ARR o mais resistente. Nesse estudo nós analisamos os polimorfismos de 15 códons diferentes da gene PrPsc em ovinos de um rebanho da raça Suffolk no Brasil afetado com scrapie clássico. Os amplicons do gene da PrPsc, que contem os códons mais frequentemente encontrados foram sequenciados para determinar o genótipo de cada animal. Nós encontramos 3 polimorfismos do 15 códons analisados (136, 143 e 171). O códon que mais teve variações foi o códon 171, onde todos os alelos foram identificados. Um polimorfismo raro foi encontrado no códon 143, em 4 por cento das amostras analisadas, o qual tem sido descrito por aumentar a resistência a scrapie em animais suscetíveis. Nenhum outro polimorfismo foi detectado nos 12 códons restantes, todos então, correspondendo à proteína priônica selvagem. De acordo com a grau de risco a desenvolver scrapie, a maioria dos animais (96 por cento) tiveram genótipo correspondentes aos grupos de risco 1 a 3 (muito baixo a moderado), e somente 4 por cento no grupo de risco alto. Nossos dados discutem a relação das medidas de prevenção envolvendo a genotipagem e a seleção positiva para o controle da doença.(AU)

Alterações clínico-laboratoriais em transtornos renais de cães (Canis familiaris) / Clinical and laboratory alterations in renal disorders of dogs (Canis familiaris)

A síndrome de insuficiência renal acarreta sinais clínicos característicos nos pacientes, bem como alterações bioquímicas hematológicas e urinárias, detectáveis pelo uso de diversos recursos laboratoriais. O presente trabalho teve por objetivo analisar dados clínicos e laboratoriais de cães atendidos com diagnóstico de transtornos renais em um Hospital de Clínicas Veterinárias. Dezesseis cães adultos (média de 6,16 anos) foram estudados. Sinais clínicos como anorexia, prostração, algia abdominal e desidratação, dentre outros, mostraram-se mais freqüentes do que sinais sugestivos de afecção renal, como poliúria, polidipsia e oligúria. O perfil hematológico destes cães revelou anemia normocítica-normocrônica, acompanhada de arcada leucocitose. O perfil bioquímico do sangue mostrou azotemia e hipoalbuminemia marcadas. A urinálise revelou densidade abaixo do normal, bem como presença de proteínas, glicose, nitritos, hemoglobina e pigmentos biliares, associados a diferentes elementos celulares e cilindros no sedimento urinário. A análise criteriosa dos resultados dos exames laboratoriais, juntamente com a amamnese e o exame clínico, fornecem ótimo subsídio para um correto diagnóstico da afecção renal(AU)

Renal insufficiency syndrome results in characteristic clinical signals and biochemical, hematological and urinary abnormalities, detectable by laboratory means. The present work aims to compile clinical and laboratory data from dogs with renal disorder diagnosis examined at a Veterinary Hospital in Southern Brazil. Sixteen adult dogs were studied. Clinical signals, like unappetency, prostration, abdominal pain and dehydration were more frequent than specific signs of renal disorders, like poliuria, polidipsia and oligúria. Hematologic profiles showed normocytic-normocromic anemia accompanied by marked leukocytosis. The blood biochemical profile showed increase in plasma urea creatinina concentrations and hipoalbuminemia. Urinalysis showed low urine density and presence of proteins, glucose, nitrites, hemoglobin and bile pigments, with different elements in the sediment. A careful analysis of laboratory results, joined to a good anamnesis and clinical examination, is the best resource for a correct diagnosis of renal disorders(AU)

Alterações clínico-laboratoriais em transtornos renais de cães (Canis familiaris) / Clinical and laboratory alterations in renal disorders of dogs (Canis familiaris)

A síndrome de insuficiência renal acarreta sinais clínicos característicos nos pacientes, bem como alterações bioquímicas hematológicas e urinárias, detectáveis pelo uso de diversos recursos laboratoriais. O presente trabalho teve por objetivo analisar dados clínicos e laboratoriais de cães atendidos com diagnóstico de transtornos renais em um Hospital de Clínicas Veterinárias. Dezesseis cães adultos (média de 6,16 anos) foram estudados. Sinais clínicos como anorexia, prostração, algia abdominal e desidratação, dentre outros, mostraram-se mais freqüentes do que sinais sugestivos de afecção renal, como poliúria, polidipsia e oligúria. O perfil hematológico destes cães revelou anemia normocítica-normocrônica, acompanhada de arcada leucocitose. O perfil bioquímico do sangue mostrou azotemia e hipoalbuminemia marcadas. A urinálise revelou densidade abaixo do normal, bem como presença de proteínas, glicose, nitritos, hemoglobina e pigmentos biliares, associados a diferentes elementos celulares e cilindros no sedimento urinário. A análise criteriosa dos resultados dos exames laboratoriais, juntamente com a amamnese e o exame clínico, fornecem ótimo subsídio para um correto diagnóstico da afecção renal

Renal insufficiency syndrome results in characteristic clinical signals and biochemical, hematological and urinary abnormalities, detectable by laboratory means. The present work aims to compile clinical and laboratory data from dogs with renal disorder diagnosis examined at a Veterinary Hospital in Southern Brazil. Sixteen adult dogs were studied. Clinical signals, like unappetency, prostration, abdominal pain and dehydration were more frequent than specific signs of renal disorders, like poliuria, polidipsia and oligúria. Hematologic profiles showed normocytic-normocromic anemia accompanied by marked leukocytosis. The blood biochemical profile showed increase in plasma urea creatinina concentrations and hipoalbuminemia. Urinalysis showed low urine density and presence of proteins, glucose, nitrites, hemoglobin and bile pigments, with different elements in the sediment. A careful analysis of laboratory results, joined to a good anamnesis and clinical examination, is the best resource for a correct diagnosis of renal disorders