RESUMO
This study describes an outbreak of alopecia in laboratory rats caused by the saprophytic fungi Scopulariopsis brevicaulis in a Brazilian lab animal facility. Initially, lesions were identified only in females during the lactation period, which showed circumscribed areas with alopecia without pruritus extensively at the abdomen and the medial side of the thighs. Later during the course of the infection, skin lesions affected both genders reaching about 10% of the colony. Skin scrapings, hair, and environmental samples from the facility were sent for analysis. The cultures were carried out in Sabouraud agar and over seven days fast-growing colonies varying in color from white to brown were observed. Micromorphology showed mycelium and characteristic ringworms of the fungus S. brevicaulis. There is a similar report of an outbreak of this fungus in Turkey. To date, this is the first report in the literature of the fungus S. brevicaulis causing disease in laboratory rats in Brazil.
Assuntos
Animais , Ratos , Surtos de Doenças/veterinária , Alopecia/veterinária , Alopecia/epidemiologia , Scopulariopsis , Animais de Laboratório/microbiologia , Ratos Wistar , AbdomeRESUMO
Trypanosoma cruzi, the etiologic agent of Chagas disease, has a complex life cycle in which four distinct developmental forms alternate between the insect vector and the mammalian host. It is assumed that replicating epimastigotes present in the insect gut are not infective to mammalian host, a paradigm corroborated by the widely acknowledged fact that only this stage is susceptible to the complement system. In the present work, we establish a T. cruzi in vitro and in vivo epimastigogenesis model to analyze the biological aspects of recently differentiated epimastigotes (rdEpi). We show that both trypomastigote stages of T. cruzi (cell-derived and metacyclic) are able to transform into epimastigotes (processes termed primary and secondary epimastigogenesis, respectively) and that rdEpi have striking properties in comparison to long-term cultured epimastigotes: resistance to complement-mediated lysis and both in vitro (cell culture) and in vivo (mouse) infectivity. Proteomics analysis of all T. cruzi stages reveled a cluster of proteins that were up-regulated only in rdEpi (including ABC transporters and ERO1), suggesting a role for them in rdEpi virulence. The present work introduces a new experimental model for the study of host-parasite interactions, showing that rdEpi can be infective to the mammalian host.
Assuntos
Doença de Chagas/parasitologia , Trypanosoma cruzi/crescimento & desenvolvimento , Trypanosoma cruzi/patogenicidade , Animais , Diferenciação Celular/fisiologia , Interações Hospedeiro-Parasita , Estágios do Ciclo de Vida/fisiologia , Camundongos , Proteínas de Protozoários/metabolismo , Trypanosoma cruzi/genética , Trypanosoma cruzi/metabolismoRESUMO
BACKGROUND: We report the isolation and characterization of dengue virus (DENV) serotype 4 from a resident of Santa Fé, state of Paraná, South Brazil, in March 2013. This patient presented with hemorrhagic manifestations, high viral load and, interestingly, a mixed Th1/Th17 cytokine profile. CASE PRESENTATION: The patient presented with classical dengue symptoms, such as fever, rash, myalgia, arthralgia, and hemorrhagic manifestations including petechiae, gum bleeding and a positive tourniquet test result. A serum sample obtained 1 day after the initial appearance of clinical symptoms was positive for NS1 viral antigen, but this sample was negative for both IgM and IgG against DENV. Dengue virus infection was confirmed by isolation of the virus from C6/36 cells, and dengue virus serotyping was performed via one-step RT-PCR. The infection was confirmed to be caused by a serotype 4 dengue virus. Additionally, based on multiple alignment and phylogeny analyses of its complete genome sequence, the viral strain was classified as genotype II (termed LRV13/422). Moreover, a mixed Th1/Th17 cytokine profile was detected in the patient's serum, and this result demonstrated significant inflammation. Biological characterization of the virus via in vitro assays comparing LRV13/422 with a laboratory-adapted reference strain of dengue virus serotype 4 (TVP/360) showed that LRV13/422 infects both vertebrate and invertebrate cell lines more efficiently than TVP/360. However, LRV13/422 was unable to inhibit type I interferon responses, as suggested by the results obtained for other dengue virus strains. Furthermore, LRV13/422 is the first completely sequenced serotype 4 dengue virus isolated in South Brazil. CONCLUSION: The high viral load and mixed Th1/Th17 cytokine profile observed in the patient's serum could have implications for the development of the hemorrhagic signs observed, and these potential relationships can now be further studied using suitable animal models and/or in vitro systems.
Assuntos
Citocinas/sangue , Vírus da Dengue/isolamento & purificação , Dengue/patologia , Dengue/virologia , Genótipo , Sorogrupo , Carga Viral , Animais , Brasil , Linhagem Celular , Análise por Conglomerados , Vírus da Dengue/classificação , Vírus da Dengue/genética , Humanos , Invertebrados , Masculino , Pessoa de Meia-Idade , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Células Th1/imunologia , Células Th17/imunologia , Vertebrados , Cultura de VírusRESUMO
Dengue is the most prevalent human arboviral disease. The morbidity related to dengue infection supports the need for an early, quick and effective diagnostic test. Brazil is a hotspot for dengue, but no serological diagnostic test has been produced using Brazilian dengue virus isolates. This study aims to improve the development of immunodiagnostic methods for dengue virus (DENV) detection through the production and characterization of 22 monoclonal antibodies (mAbs) against Brazilian isolates of DENV-1, -2 and -3. The mAbs include IgG2bκ, IgG2aκ and IgG1κ isotypes, and most were raised against the envelope or the pre-membrane proteins of DENV. When the antibodies were tested against the four DENV serotypes, different reactivity patterns were identified: group-specific, subcomplex specific (DENV-1, -3 and -4 and DENV-2 and -3) and dengue serotype-specific (DENV-2 or -3). Additionally, some mAbs cross-reacted with yellow fever virus (YFV), West Nile virus (WNV) and Saint Louis encephalitis virus (SLEV). None of the mAbs recognized the alphavirus Venezuelan equine encephalitis virus (VEEV). Furthermore, mAbs D3 424/8G, D1 606/A12/B9 and D1 695/12C/2H were used to develop a capture enzyme-linked immunosorbent assay (ELISA) for anti-dengue IgM detection in sera from patients with acute dengue. To our knowledge, these are the first monoclonal antibodies raised against Brazilian DENV isolates, and they may be of special interest in the development of diagnostic assays, as well as for basic research.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Vírus da Dengue/imunologia , Dengue/imunologia , Animais , Brasil , Linhagem Celular , Reações Cruzadas , Dengue/diagnóstico , Dengue/virologia , Vírus da Dengue/classificação , Epitopos/imunologia , Humanos , Imunoglobulina G/imunologia , CamundongosRESUMO
Com o advento da promulgação da Lei 11794/2008 (Lei Arouca), posteriormente regu- lamentada pelo Decreto 6899/2009, iniciou-se uma corrida para a implementação de mudanças no cenário da experimentação animal no Brasil, com ênfase para a neces- sidade de melhorias no gerenciamento dos biotérios. Com o intuito de facilitar essas melhorias, o uso de softwares específicos para o gerenciamento de biotérios tornou-se uma opção atrativa para a manutenção e recuperação dos dados das diferentes rotinas do criatório animal. Estes programas proporcionam melhor visualização da alocação dos recursos financeiros e facilitam tomadas de decisões gerenciais mais rápidas e seguras. Com o objetivo de melhorar a capacidade do processo gerencial do nosso biotério, desenvolvemos e implementamos um software baseado em linguagens de acesso livre, denominado BioterC. As etapas de desenvolvimento e implementação do software foram fundamentadas na interação direta entre o desenvolvedor e os bioteristas, propiciando a construção de uma ferramenta robusta, versátil, de uso intuitivo, em um conceito de elevada adaptabilidade e flexibilidade, sem qualquer investimento adicional com hardware por parte do biotério. Uma vez colocado em uso, o programa pôde ser posteriormente adaptado, visando à adequação do software com a realidade apresentada na rotina de cada ambiente do biotério. No presente relatório técnico descrevemos o desenvolvimento e a implementação de um software baseado em linguagens de programação de acesso livre, utilizado como uma ferramenta gerencial aplicada a biotérios.(AU)
The publication of Federal Brazilian Law 11.794/2008 (also named Law Arouca), regulated by Decree 6899/2009, was the major circumstance that supported changes in laboratory animal science in Brazil, in special those related to management control. The lack of management into lab animal facilities is related main to a limited capacity of both data recording and data recovery. Thus, softwares specifically developed to Furthermore, it allows a better overview of financial resources allocation and can maintain and analyze lab animal facilities data helping the overall data management. be used to take fast management decisions by lab animal staff. In order to improve the management in our lab animal facility was developed and implemented a free language-based software, termed BioterC. The whole software programming was based on close interaction between programmer developer and lab animal staff. This methodology allowed the construction of a robust, versatile, use intuitive, on a concept of high adaptability and flexibility software, without any additional investment in hardware by the lab animal facility. When software began to be used it could be constantly adapted to reality found into each routine from the lab animal facility. In this technical report we describe the developing and implementing of a free language software-based used as management tool applied in lab animal facilities.(AU)
Assuntos
Animais , Legislação Veterinária/tendências , Software/tendências , Animais de Laboratório/classificaçãoRESUMO
Com o advento da promulgação da Lei 11794/2008 (Lei Arouca), posteriormente regu- lamentada pelo Decreto 6899/2009, iniciou-se uma corrida para a implementação de mudanças no cenário da experimentação animal no Brasil, com ênfase para a neces- sidade de melhorias no gerenciamento dos biotérios. Com o intuito de facilitar essas melhorias, o uso de softwares específicos para o gerenciamento de biotérios tornou-se uma opção atrativa para a manutenção e recuperação dos dados das diferentes rotinas do criatório animal. Estes programas proporcionam melhor visualização da alocação dos recursos financeiros e facilitam tomadas de decisões gerenciais mais rápidas e seguras. Com o objetivo de melhorar a capacidade do processo gerencial do nosso biotério, desenvolvemos e implementamos um software baseado em linguagens de acesso livre, denominado BioterC. As etapas de desenvolvimento e implementação do software foram fundamentadas na interação direta entre o desenvolvedor e os bioteristas, propiciando a construção de uma ferramenta robusta, versátil, de uso intuitivo, em um conceito de elevada adaptabilidade e flexibilidade, sem qualquer investimento adicional com hardware por parte do biotério. Uma vez colocado em uso, o programa pôde ser posteriormente adaptado, visando à adequação do software com a realidade apresentada na rotina de cada ambiente do biotério. No presente relatório técnico descrevemos o desenvolvimento e a implementação de um software baseado em linguagens de programação de acesso livre, utilizado como uma ferramenta gerencial aplicada a biotérios.
The publication of Federal Brazilian Law 11.794/2008 (also named Law Arouca), regulated by Decree 6899/2009, was the major circumstance that supported changes in laboratory animal science in Brazil, in special those related to management control. The lack of management into lab animal facilities is related main to a limited capacity of both data recording and data recovery. Thus, softwares specifically developed to Furthermore, it allows a better overview of financial resources allocation and can maintain and analyze lab animal facilities data helping the overall data management. be used to take fast management decisions by lab animal staff. In order to improve the management in our lab animal facility was developed and implemented a free language-based software, termed BioterC. The whole software programming was based on close interaction between programmer developer and lab animal staff. This methodology allowed the construction of a robust, versatile, use intuitive, on a concept of high adaptability and flexibility software, without any additional investment in hardware by the lab animal facility. When software began to be used it could be constantly adapted to reality found into each routine from the lab animal facility. In this technical report we describe the developing and implementing of a free language software-based used as management tool applied in lab animal facilities.