Ruptura idiopática de bexiga urinária em duas éguas

Urinary bladder rupture is rarely reported in adult horses and is usually associated with parturition, history of traumaor urethral obstruction. This paper reports two cases in mares free of these predisposing factors, diagnosed throughclinical manifestations, abdominal ultrasound and abdominocentesis, corrected by cystorrhaphy through laparotomy.While recovery was satisfactory in one case, the other case had complications in celiorraphy such as development ofincisional hernia, small colon eventration and intestinal adherences, opting by the euthanasia of the animal. These reportsare important because of the rarity with which they occur in adult mares. The different prognoses may be related to theseverity of each case, according to the location and extension of the wound, which directly or indirectly influence thetechnique and recovery of the animals.A ruptura de bexiga urinária é raramente relatada em equinos adultos, sendo normalmente associada ao parto, antecedentesde trauma ou obstrução de uretra. Este trabalho relata dois casos em éguas livres destes fatores predisponentes,diagnosticados através de manifestações clínicas, ultrassonografia abdominal e abdominocentese, corrigidos por cistorrafiaatravés de laparotomia. Em um dos casos a recuperação ocorreu satisfatoriamente, porém no outro houve complicaçõesna celiorrafia como desenvolvimento de hérnia incisional, eventração de cólon menor e posterior desenvolvimento deaderências intestinais, optando-se pela eutanásia do animal. Estes relatos são importantes pela raridade com que ocorremem éguas adultas. Os diferentes prognósticos podem estar relacionados à gravidade de cada caso, de acordo com o locale extensão da ferida, que direta ou indiretamente influenciam na técnica empregada e recuperação dos animais

Development and application of a real-time RT-PCR for brazilian Vesiculovirus

Os Vesiculovirus são um gênero de vírus de RNA da família Rhabdoviridae que inclui os sorotipos Carajás, Cocal, Marabá, Piry, Alagoas e Indiana. Estes são causadores de estomatite vesicular em ruminantes e doença febril humana no Brasil. As vesiculoviroses e suas epidemiologias são pouco conhecidas em seres humanos. Ainda, os Vesiculovirus (VSV) são pouco diagnosticados no homem e em animais pela escassez de métodos laboratoriais diagnósticos. Por isso, objetivamos neste trabalho desenvolver e testar uma RT-PCR em tempo real, pelo método SYBR Green, visando à detecção de VSV brasileiros. Primers que amplificam parte do gene da proteína G dos VSV foram utilizados no teste o qual mostrou-se capaz de detectar genomas dos VSV Piry, Indiana, Alagoas e Carajás. O método foi usado para testar amostras séricas de pacientes com doença febril aguda, de bovinos, de equinos e de macerados de artrópodos. A RT-PCR em tempo real mostrou-se 100 vezes mais sensível que a RT-PCR convencional para Vesiculovirus e também, permitiu detectar até 10 cópias de RNA do vírus Piry. Ainda, a RT-PCR em tempo real para Vesiculovirus mostrou-se capaz de diagnosticar e quantificar o VSV Alagoas nas amostras séricas de bovinos e de equinos. Portanto, a RT-PCR em tempo real desenvolvida neste trabalho, provavelmente, será muito útil no diagnóstico e também, em futuras pesquisas, que permitirão ampliar o conhecimento epidemiológico, ainda pouco conhecido, sobre os Vesiculovirus.The Vesiculovirus is a Rhabdoviridae family genre of RNA virus that includes serotypes Carajás, Cocal, Maraba, Piry, Alagoas and Indiana. These are causes of vesicular stomatitis in ruminants and human febrile illness in Brazil. The vesiculoviroses and its epidemiology are little known in humans. Still, Vesiculovirus (VSV) are poorly diagnosed in humans and laboratory animals by the lack of diagnostic methods. Therefore, we proposed in this work to develop and test a real-time RT-PCR by SYBR Green method, focusing on the detection of Brazilian VSV. Primers that amplify part of the VSV G protein gene were used in the test which proved capable of detecting genomes of VSV Piry, Indiana, Alagoas and Carajás. The method was used to test serum samples from patients with acute febrile disease, cattle, horses and macerated arthropods. Real time RT-PCR showed to be 100 times more sensitive than conventional RT-PCR for Vesiculovirus and also was possible to detect up to 10 RNA copies of the Piry virus. Also, the real-time RT-PCR for Vesiculovirus proved able to diagnose and quantify Alagoas VSV in serum samples from cattle and horses. Therefore, the real-time RT-PCR developed in this work will probably be very useful in the diagnosis and in future research, which will increase the epidemiological knowledge, as it is still little known about the Vesiculovirus

Research on compounds with potential antiviral action for Mayaro and Oropouche viruses

Os Alphavirus e Orthobunyavirus são arbovírus importantes que causam grande impacto econômico e social no Brasil. Não existem antivirais disponíveis para o tratamento desses vírus. Portanto, é fundamental ter medicamentos terapêuticos que combatam os sintomas e sinais manifestados pela doença em casos leves e moderados. Neste trabalho, desenvolvemos um ensaio de luminescência baseada em células para a triagem de potenciais pequenas moléculas para os vírus Mayaro e Oropouche que mede o efeito citopático (CPE) induzido pelo vírus em células Vero usando o sistema CellTiter Glo baseado em luminescência. O ensaio foi validado no formato de placa de 384 poços e mostrou valores de Z maiores que 0,7, background maior que 30 e sinal-ruído maior que 10, demonstrando alta capacidade. Quatro bibliotecas de compostos foram testadas na concentração de 10 uM para ambos os testes. Identificamos cinco compostos que inibiram o efeito citopático induzido pelos vírus em > 50%, com valores de EC50/CC50 comparáveis aos determinados por outros ensaios baseados em células, validando assim a precisão e a capacidade do ensaio de servir como uma ferramenta para a descoberta de novos antivirais para os vírus Mayaro e Oropouche.Alphavirus and Orthobunyavirus are important arboviruses that have a major economic and social impact in Brazil. There are no antivirals available to treat these viruses. Therefore, it is essential to have therapeutic drugs that combat the symptoms and signs manifested by the disease in mild and moderate cases. In this thesis, we developed a cell-based luminescence assay for the screening of potential small molecules for Mayaro and Oropouche viruses that measures the virus-induced cytopathic effect (CPE) in Vero cells (ATCC CCL81) using the luminescence-based CellTiter Glo system. The assay was validated in a 384-well plate format and showed Z values greater than 0.7, background greater than 30 and signal-to-noise greater than 10, demonstrating high capacity. Four compound libraries were tested at a concentration of 10 µM for both tests. We identified five compounds that inhibited virus-induced cytopathic effect by> 50%, with EC50/CC50 values comparable to those determined by other cell-based assays, thus validating the assay\'s accuracy and ability to serve as a tool for discovery of new antivirals for Mayaro and Oropouche viruses

Molecular Characterization Of Capim And Enseada Orthobunyaviruses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Capim and Enseada viruses are members of the genus Orthobunyavirus isolated from mosquitoes and mammals in Brazil. Despite seroprevalence studies indicating human infections in Latin America, these viruses remain relatively unknown and unstudied. In order to better understand the genetic and evolutionary relationships among orthobunyaviruses, we sequenced the three genomic segments of Capim and Enseada orthobunyaviruses. Based on phylogenetic analysis, we demonstrated that these viruses depicted two new distinct clades, one represented by Enseada and another composed of Capim virus. In general, the genome organization and genetic traits of these viruses are similar to other orthobunyaviruses however, the open reading frame (ORF) of the putative nonstructural NSs protein of Enseada orthobunyavirus precedes the nucleocapsid ORF. Overall, our study provides details on the molecular characteristics of the prototype species of two groups within the Orthobunyavirus genus, revealing novel features into the genetic diversity and evolution of this genus. (c) 2016 Elsevier B.V. All rights reserved.404753Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Brazil [13/14929-1, 14/02438-6, 12/24150-9, 15/05778-5, 14/20851-8]CNPq [302032/2011-8, 200024/2015-9]CNPq 1B senior fellowship [301677/2013-1]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Complete Genome Sequence Of Piry Vesiculovirus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Piry virus (PIRYV) is a rhabdovirus (genus Vesiculovirus) and is described as a possible human pathogen, originally isolated from a Philander opossum trapped in Para State, Northern Brazil. This study describes the complete full coding sequence and the genetic characterization of PIRYV. The genome sequence reveals that PIRYV has a typical vesiculovirus-like organization, encoding the five genes typical of the genus. Phylogenetic analysis confirmed that PIRYV is most closely related to Perinet virus and clustered in the same clade as Chandipura and Isfahan vesiculoviruses.161823252328Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Brazil [13/14929-1, 14/02438-6, 12/24150-9, 15/05778-5, 14/20851-8, 13/02256-2]CNPq [302032/2011-8, 200024/2015-9]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Evaluation and optimization of SYBR Green real-time reverse transcription polymerase chain reaction as a tool for diagnosis of the Flavivirus genus in Brazil

Abstract: INTRODUCTION: The genus Flavivirus includes several pathogenic species that cause severe illness in humans. Therefore, a rapid and accurate molecular method for diagnosis and surveillance of these viruses would be of great importance. Here, we evaluate and optimize a quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) method for the diagnosis of the Flavivirus genus. METHODS: We evaluated different commercial kits that use the SYBR Green system for real-time RT-PCR with a primer set that amplifies a fragment of the NS5 flavivirus gene. The specificity and sensitivity of the assay were tested using twelve flaviviruses and ribonucleic acid (RNA) transcribed from the yellow fever virus. Additionally, this assay was evaluated using the sera of 410 patients from different regions of Brazil with acute febrile illness and a negative diagnosis for the dengue virus. RESULTS: The real-time RT-PCR amplified all flaviviruses tested at a melting temperature of 79.92 to 83.49°C. A detection limit of 100 copies per ml was determined for this assay. Surprisingly, we detected dengue virus in 4.1% (17/410) of samples from patients with febrile illness and a supposedly negative dengue infection diagnosis. The viral load in patients ranged from 2.1×107to 3.4×103copies per ml. CONCLUSIONS: The real-time RT-PCR method may be very useful for preliminary diagnoses in screenings, outbreaks, and other surveillance studies. Moreover, this assay can be easily applied to monitor viral activity and to measure viral load in pathogenesis studies

Molecular characterization of capim and enseada orthobunyaviruses

Capim and Enseada viruses are members of the genus Orthobunyavirus isolated from mosquitoes and mammals in Brazil. Despite seroprevalence studies indicating human infections in Latin America, these viruses remain relatively unknown and unstudied. In order to better understand the genetic and evolutionary relationships among orthobunyaviruses, we sequenced the three genomic segments of Capim and Enseada orthobunyaviruses. Based on phylogenetic analysis, we demonstrated that these viruses depicted two new distinct clades, one represented by Enseada and another composed of Capim virus. In general, the genome organization and genetic traits of these viruses are similar to other orthobunyaviruses however, the open reading frame (ORF) of the putative nonstructural NSs protein of Enseada orthobunyavirus precedes the nucleocapsid ORF. Overall, our study provides details on the molecular characteristics of the prototype species of two groups within the Orthobunyavirus genus, revealing novel features into the genetic diversity and evolution of this genus404753CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP302032/2011-8; 200024/2015-9; 301677/2013-113/14929-1; 14/02438-6; 12/24150-9; 15/05778-5; 14/20851-

Molecular characterization of Capim and Enseada orthobunyaviruses

Capim and Enseada viruses are members of the genus Orthobunyavirus isolated from mosquitoes and mammals in Brazil. Despite seroprevalence studies indicating human infections in Latin America, these viruses remain relatively unknown and unstudied. In order to better understand the genetic and evolutionary relationships among orthobunyaviruses, we sequenced the three genomic segments of Capim and Enseada orthobunyaviruses. Based on phylogenetic analysis, we demonstrated that these viruses depicted two new distinct clades, one represented by Enseada and another composed of Capim virus. In general, the genome organization and genetic traits of these viruses are similar to other orthobunyaviruses however, the open reading frame (ORF) of the putative nonstructural NSs protein of Enseada orthobunyavirus precedes the nucleocapsid ORF. Overall, our study provides details on the molecular characteristics of the prototype species of two groups within the Orthobunyavirus genus, revealing novel features into the genetic diversity and evolution of this genus

Complete genome sequence of Piry vesiculovirus

Piry virus (PIRYV) is a rhabdovirus (genus Vesiculovirus) and is described as a possible human pathogen, originally isolated from a Philander opossum trapped in Para State, Northern Brazil. This study describes the complete full coding sequence and the genetic characterization of PIRYV. The genome sequence reveals that PIRYV has a typical vesiculovirus-like organization, encoding the five genes typical of the genus. Phylogenetic analysis confirmed that PIRYV is most closely related to Perinet virus and clustered in the same clade as Chandipura and Isfahan vesiculoviruses161823252328CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP302032/2011-8; 200024/2015-913/14929-1; 14/02438-6; 12/24150-9; 15/05778-5; 14/20851-8; 13/02256-

Sequência completa do genoma de Piry vesiculovirus

University of São Paulo. School of Medicine of Ribeirão Preto. Virology Research Center. Ribeirão Preto, SP, Brazil.University of Sao Paulo. School of Medicine. Department of Cell and Molecular Biology. Ribeirão Preto, SP, Brazil.University of São Paulo. School of Medicine of Ribeirão Preto. Virology Research Center. Ribeirão Preto, SP, Brazil.Universidade Estadual de Campinas. Laboratório Central de Tecnologias de Alto Desempenho em Ciências da Vida. Campinas, SP, Brazil.University of São Paulo. School of Medicine of Ribeirão Preto. Virology Research Center. Ribeirão Preto, SP, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.University of São Paulo. School of Medicine of Ribeirão Preto. Virology Research Center. Ribeirão Preto, SP, Brazil.Piry virus (PIRYV) is a rhabdovirus (genus Vesiculovirus) and is described as a possible human pathogen, originally isolated from a Philander opossum trapped in Para State, Northern Brazil. This study describes the complete full coding sequence and the genetic characterization of PIRYV. The genome sequence reveals that PIRYV has a typical vesiculovirus-like organization, encoding the five genes typical of the genus. Phylogenetic analysis confirmed that PIRYV is most closely related to Perinet virus and clustered in the same clade as Chandipura and Isfahan vesiculoviruses