Made available in DSpace on 2016-04-28T20:15:28Z (GMT). No. of bitstreams: 1 2007- Ana Luisa Perrone Poerner.pdf: 1459894 bytes, checksum: 00367d6cd6bb2110295c0fa6aad7546d (MD5) Previous issue date: 2007-02-22 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The objectives of this study were to understand the characteristics of human rabies postexposure prophylaxis (PEP) in Center-South Region of Rio de Janeiro State and analyze PEP trends against epidemiologic situation of canine rabies, indirectly analyzing integration between control actions developed in the area. Data from 4310 patients who visited health services because of animal aggressions between years 2000 and 2005 were obtained in Health State Department, as well as canine vaccination coverage information. In this paper, descriptive analysis was used to study variables like: method and source of exposure occurrence, patients age and sex, health conditions and species of the animal aggressor, PEP interruption, characteristics of the wounds and previously PEP prescription. Data related to animal exposure incidences, PEP prescription incidence, dogs density, dog/human rate and canine vaccination coverage were also studied. Simple linear regression was carried out to obtain human rabies prescription and canine vaccination trends. c? test was used to verify significant difference (r ? 0,05) among vaccinated patients proportion in the period. Incidence of animal agression ranged from 10,1/10.000 persons in 2000 to 29,2/10.000 persons in 2005, with an average of 23,7/10.000 persons. Bite exposures accounted for 79,3% of the wounds, dogs were responsible for 86,2% of the cases and most of them were healthy at the moment of the aggression (44,8%). Male patients accounted for 57,5% of the cases. Most of the patients between 1 and 15 years (35%) also were male (62,2%). Most of the wounds were unique (59,7%) superficial (60,8%) and not-provoked (60,1%). Incidence of rabies PEP ranged from 5,1/10.000 persons in 2000 to 23,6/10.000 persons in 2005, reaching an average of 15,2/10.000 persons. Rabies PEP prescription average and estimated canine vaccination coverage in the period was 64,1% and 92,8%, respectively. Estimated relation dog/human in 2005 was 1:5,8 and dog density in the same year was 16,7 dogs/km?. There was significant difference between proportions of vaccinated and non-vaccinated patients in the period. The characteristics of human rabies post-exposure prophylaxis and animal aggressions hadn t changed and there was an ascended trend in human rabies post-exposure prophylaxis prescription, with high proportions and significant difference, despite the high levels of canine vaccination coverage and absence of human and canine rabies notification, indicating a lack of integration between prevention actions developed in the area. / O presente estudo teve como objetivo conhecer as caracter?sticas dos casos de atendimento anti-r?bico humano p?s-exposi??o na Regi?o Centro-Sul do Estado do Rio de Janeiro e analisar a tend?ncia da prescri??o de vacina anti-r?bica a humanos, v?timas de agress?o animal, frente ? situa??o epidemiol?gica da raiva canina, avaliando indiretamente a integra??o das a??es de profilaxia da raiva animal e humana na Regi?o. Os dados referentes ao atendimento anti-r?bico humano p?s-exposi??o de 4310 pacientes e informa??es sobre a cobertura vacinal canina, no per?odo de 2000 a 2005, foram obtidos na Secretaria Estadual de Sa?de. Foi realizada uma an?lise descritiva das vari?veis: tipo e modo de ocorr?ncia da agress?o, esp?cie animal agressora e sua condi??o, circunst?ncias da agress?o, localiza??o, gravidade e caracter?sticas do ferimento, motiva??o para a ocorr?ncia das agress?es, faixa et?ria e g?nero dos indiv?duos expostos, hist?rico de profilaxia anti-r?bica dos pacientes e ocorr?ncia de interrup??o da vacina??o. Foram calculadas a incid?ncia de atendimento antir?bico humano p?s-exposi??o, incid?ncia e percentual de prescri??o de vacina anti-r?bica humana p?s-exposi??o, densidade populacional de c?es, raz?o c?o/homem e cobertura vacinal canina. A regress?o linear simples foi utilizada para obten??o das retas de tend?ncia da prescri??o de vacina anti-r?bica humana e da vacina??o canina. O teste do c? foi utilizado para verificar se a propor??o de pacientes vacinados diferiu significativamente (r ? 0,05) entre os anos analisados. A incid?ncia de atendimentos variou de 10,1/10.000 habitantes em 2000 a 29,2/10.000 habitantes em 2005, com uma incid?ncia m?dia de atendimentos de 23,7/10.000 habitantes. A mordedura foi o principal tipo de agress?o (79,3%) e os c?es foram os principais agressores (86,2%), sendo que a maioria (44,8%) se encontrava sadia no momento da agress?o. A maioria dos pacientes (57,5%) foi do g?nero masculino. Os indiv?duos mais expostos tinham entre 1 e 15 anos de idade (35%), sendo que a maioria tamb?m era do sexo masculino (62,2%). A maior parte das les?es localizou-se nas m?os e p?s (36,6%), sendo que em crian?as menores de 15 anos a maioria das les?es ocorreu na cabe?a e pesco?o (75,6%). Predominaram os ferimentos ?nicos (59,7%), superficiais (60,8%) e n?o-provocados (60,1%). A incid?ncia de prescri??o de vacina anti-r?bica humana variou de 5,1/10.000 habitantes em 2000 a 23,6/10.000 habitantes em 2005, com uma incid?ncia m?dia de prescri??o de vacina de 15,2/10.000 habitantes. O percentual m?dio de pessoas vacinadas foi de 64,1%. A m?dia de cobertura vacinal canina atingida na regi?o foi de 92,8%. A rela??o c?o/homem estimada na regi?o em 2005 foi de 1:5,8 e a densidade populacional canina no mesmo ano foi de 16,7 c?es/km?. A propor??o de pacientes vacinados e n?o vacinados diferiu significativamente no per?odo analisado. As caracter?sticas dos casos de atendimento e dos animais agressores n?o se alteraram em rela??o aos relatados na literatura e observou-se uma tend?ncia ascendente na prescri??o de vacina anti-r?bica humana p?s-exposi??o, com altos percentuais e diferen?as significativas, apesar de altos n?veis de cobertura vacinal canina e aus?ncia de notifica??o de raiva canina e humana, demonstrando uma falta de articula??o entre as a??es do programa de controle na Regi?o.

Made available in DSpace on 2016-04-28T20:15:36Z (GMT). No. of bitstreams: 1 2009 - Julio Cesar Augusto Pompei.pdf: 279592 bytes, checksum: d5618460bc231e06151836a53b7af00a (MD5) Previous issue date: 2009-12-15 / The National Program on Control of Herbivore Rabies (PNCRH) concentrates its actions on population control of the disease s main transmitter, the vampire bat of the Desmodus rotundus species, and favors the use of direct and indirect selective methods. This study sought to evaluate the effectiveness of the VAMPIRICID GEL? for direct selective control of D. rotundus colonies. Six daytime shelters were selected; in each one a count was taken of the live individuals at the time and then again ten days after treatment. Two shelters per treatment Group were addressed. The Vampiricid Gel was applied on the back of vampire bats in 5-percent (Group 1) and of 10-percent (Group 2) samples of the population. For control (Group 3), 10 percent of the D. rotundus bat population was treated with liquid Vaseline. An evaluation was also made of the aggressions by vampire bats on 3,690 bovines and 133 equids on the 22 properties located within a 3-km radius of the shelters. Fisher s Exact and χ2 tests were used with a 1-percent significance level. Under natural conditions, treatment of Groups 1, 2, and 3 reduced by 97.57 percent, 99.30 percent, and 3.6 percent, respectively, the D. rotundus population, and by 92.79 percent, 94.36 percent, and 4.76 percent, respectively, the number of animals attacked, ten days after treatment. Results have shown that the 1-percent VAMPIRICID GEL ? was effective in significantly reducing the bat population and the number of animals attacked in Groups 1 and 2, as compared with Group 3. There was no significant different between Groups 1 and 2 as regards both reduction of the D. rotundus population and the number of animals attacked. Although the use of 1-percent VAMPIRICID GEL ? on 5 percent or 10 percent of the vampire bat population has yielded similar results in the D. rotundus populations in the shelters and thus on attacked bovines and equids, its use on 5 percent of the populations was more effective, given the lower operating cost, and can thus be indicated for direct selective control of D. rotundus. / O Programa Nacional de Controle da Raiva dos Herb?voros (PNCRH) concentra suas a??es no controle populacional de seu principal transmissor, o morcego hemat?fago da esp?cie Desmodus rotundus preconizando a utiliza??o de m?todos seletivos diretos e indiretos. O presente estudo teve como objetivo avaliar a efici?ncia do VAMPIRICID GEL? no controle seletivo direto de col?nias de D. rotundus. Foram selecionados seis abrigos diurnos e em cada um foi realizada a contagem do n?mero de indiv?duos vivos no momento e dez dias ap?s os tratamentos. Utilizaram-se dois abrigos por grupo de tratamento. O gel vampiricida foi aplicado no dorso dos morcegos hemat?fagos em amostras de 5% (grupo 1) e de 10% (grupo 2) da popula??o. Para controle (grupo 3) 10% da popula??o de morcegos D. rotundus foi tratada com vaselina l?quida. Avaliou-se tamb?m a redu??o de agress?es por morcegos hemat?fagos em 3690 bovinos e 133 eq??deos existentes nas 22 propriedades localizadas num raio de 3 km de dist?ncia dos abrigos. Os testes do χ2 e Fisher Exato foram utilizados com n?vel de signific?ncia de 1%. Em condi??es naturais, os tratamentos nos grupos 1, 2 e 3 reduziram em 97,57, 99,30 e 3,6% a popula??o de D. rotundus e em 92,79, 94,36 e 4,76% o n?mero de animais agredidos, respectivamente, 10 dias ap?s os tratamentos. Os resultados obtidos demonstraram que o produto VAMPIRICID GEL ? 1% foi capaz de reduzir significativamente a popula??o de morcegos e o n?mero de animais agredidos nos grupos 1 e 2 quando comparados ao grupo 3. Entre os Grupos 1 e 2 n?o houve diferen?a significativa tanto na redu??o da popula??o de D. rotundus quanto na redu??o do n?mero de animais agredidos. Embora o uso do VAMPIRICID GEL ? 1% em 5% ou 10% da popula??o de morcegos hemat?fagos tenha produzido resultados similares na redu??o da popula??o de D. rotundus nos abrigos, e conseq?entemente do n?mero de bovinos e eq??deos agredidos, o seu uso em 5% da popula??o foi mais eficiente, devido ao menor custo operacional, podendo ser indicado para o controle seletivo direto de D. rotundus.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-09-11T18:28:18Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_ImunopatologiaExperimentalVirus.pdf: 28883093 bytes, checksum: 4d12e0ab15cafaa57569f2075ac00e53 (MD5) / Approved for entry into archive by Irvana Coutinho (irvana@ufpa.br) on 2017-09-22T15:15:34Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_ImunopatologiaExperimentalVirus.pdf: 28883093 bytes, checksum: 4d12e0ab15cafaa57569f2075ac00e53 (MD5) / Made available in DSpace on 2017-09-22T15:15:35Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_ImunopatologiaExperimentalVirus.pdf: 28883093 bytes, checksum: 4d12e0ab15cafaa57569f2075ac00e53 (MD5) Previous issue date: 2015-06-02 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / A raiva é uma zoonose por ter como hospedeiros, reservatórios e transmissores, mamíferos silvestres ou domésticos, caracterizada por doença aguda, causada pelo vírus da raiva (RABV) que compromete o sistema nervoso central, caracterizando-se por encefalite, com prognóstico fatal em quase todos os casos, em qualquer espécie de mamíferos. O objetivo deste trabalho foi descrever achados patológicos e imunopatologia de diferentes cepas de vírus da raiva nos tecidos do sistema nervoso central (SNC) verificando a resposta imunologica celular e humoral durante infecção experimental de camundongos Mus musculus. Os animais foram inoculados experimentalmente com duas variantes antigênicas do RABV (VAg2 e VAg3), por diferentes vias de infecção, e um grupo controle. Os animais foram observados quanto ao desenvolvimento de sinais clínicos e sintomas, sendo coletados e eutanasiados seguindo uma cinética. Os tecidos foram fixados em formaldeído a 10%, incluídos em blocos de parafina, corados por hematoxilina-eosina para análise histopatológica, e marcados com anticorpos específicos para imunohistoquímica a fim de caracterizar e quantificar in situ a distribuição do antígeno e a resposta inflamatória. Antígenos do RABV foram encontrados no SNC de maneira difusa, mas principalmente nos neurônios. Foi observada supressão dos linfócitos TCD4+, com aumento dos linfócitos TCD8+. Observou-se apoptose importante, com morte de células da glia. Houve aumento de citocinas pro-inflamatórias (TNF-α, IFN-γ, IL-6, IL- 1β e IL-8), anti-inflamatórias (TGF-β e IL-4) e iNOS em ambas as variantes antigênicas do RABV, mas sem observação de um perfil TH17. Esta análise possibilitou caracterizar a raiva como uma meningoencefalite, por acometer os microambientes meningeal, perivascular e intraparenquimatoso. E o processo inflamatório foi verificado mesmo quando na presençaa de corpusculos de Negri, porém com menor intensidade. / The rabies is considered a zoonosis due have as host, reservoirs and transmitters the domestic or wild mammals. It´s characterized in acute disease caused by rabies virus (RABV) that affects the central nervous system (CNS) characterized by encephalitis with fatal prognosis in almost all cases, in any mammalian species. The aim of this study was to describe pathological findings and immunopathology of different strains of rabies virus in the tissues of the central nervous system, checking cellular and humoral immune response during experimental infection of Mus musculus mice. The animals were inoculated with two antigenic variants of RABV (VAg2 and VAg3), by different routes of infection, and a control group. The animals were observed for development of clinical signs and symptoms, collected and euthanized following a kinetic. The tissues were fixed in formaldehyde 10%, embedded in paraffin, stained with hematoxylin-eosin for histopathological analysis and with specific antibodies for immunohistochemical to characterize and quantify in situ distribution of the antigen and the inflammatory response. RABV antigens were found in the CNS in a diffuse way, but mainly in neurons. It was observed suppression of CD4+ lymphocytes, with increase of CD8+ lymphocytes. It was observed significant apoptosis with glial cell death and an increase of proinflammatory cytokines (TNF-α, IFN-γ, IL-6, IL-1β and IL-8), anti-inflammatory (TGF-β and IL-4) and iNOS in both antigenic variants of RABV, but without observation of a TH17 profile. The analyses enable the characterization of rabies as meningoencephalitis, since it affects the meningeal, perivascular and intraparenchymal microenvironments. And the inflammatory process was observed even in the presence of inclusion bodies, but with less intensity.

Resumo: A intensa convivência entre o ser humano e os animais de estimação não tem sido acompanhada pela adoção de posturas de posse responsável. Dentre os animais de estimação, cães e gatos são geralmente os de eleição, estando presentes em grande contingente dos lares. Ações educativas que alertem e conscientizem sobre zoonoses e sobre a responsabilidade de possuir animais são fundamentais na mitigação dos problemas de saúde pública; porém, antes da realização de qualquer programa, é imprescindível o conhecimento da realidade local. O presente estudo foi idealizado com o propósito de avaliar a população de cães e gatos, e o nível de conhecimento e comportamento sobre a raiva e posse responsável de animais de estimação, em bairros de níveis socioeconômicos contrastantes da cidade de Jaboticabal, Estado de São Paulo. De setembro a novembro de 2007 foram entrevistados 185 moradores dos bairros Jardim Patriarca, Jardim Paulista e Jardim Santa Rosa, e 197 moradores do bairro Nova Jaboticabal, totalizando 382 entrevistas. Para análise dos dados foram empregados o teste Qui-quadrado e a Análise de Correspondência Múltipla. Apesar do contraste socioeconômico, o nível de conhecimento dos entrevistados sobre posse responsável e raiva animal foi semelhante em todos os bairros estudados; entretanto, o melhor poder aquisitivo favoreceu maiores gastos com serviços veterinários, como atendimento médico e vacinações, e maior número de animais por residência. As informações obtidas respaldam a necessidade de realizar trabalhos educativos constantes sobre posse responsável e prevenção de agravos em toda a cidade, além do estabelecimento de um canal de comunicação entre veterinários e o governo municipal. / Abstract: The intense interactions between people and pets have not been accompanied by responsible ownership practices over the years. Dogs and cats are the pets of choice, and they are found in great amount of homes. It is crucial to put into practice educative actions on zoonoses and on responsibility of owning animals to mitigate public health-related problems. However, before their effectuation, it is necessary to assess the local reality. The present study aimed to assess owned-dogs and cats populations and to evaluate the level of knowledge of responsible pet ownership and rabies among residents of socially and economically contradictory neighborhoods of the city of Jaboticabal, State of São Paulo. From September to November, 2007, 185 residents of Jardim Patriarca, Jardim Paulista and Jardim Santa Rosa neighborhoods and 197 residents of Nova Jaboticabal were interviewed, giving a total of 382 interviews. Chi-square test and Multiple Correspondence Analysis were used for data analysis. Despite socioeconomic contradiction, the level of knowledge was similar. However, better levels of income favored greater expenses with veterinary care services, including vaccinations, and a greater amount of pets per household. Results recommend the need for constant educational actions on responsible pet ownership and injury prevention for all social classes of Jaboticabal, besides the establishment of a communication channel between private veterinarians and local government. / Orientadora: Adolorata Aparecida Bianco Carvalho / Coorientador: Antonio Sérgio Ferraudo / Banca: Luiz Augusto do Amaral / Banca: Maria de Lourdes Aguiar Bonadia Reichmann / Mestre

Orientador: Adolorata Aparecida Bianco Carvalho / Coorientador: Daniel Figuglietti Brandespim / Banca: Fumio Honma Ito / Banca: Darci Lara Perecin Nocitti / Banca: Luís Antônio Mathias / Banca: Maria da Glória Buzinaro / Resumo: A raiva é uma antropozoonose de evolução letal causada por vírus do gênero Lyssavirus. É uma das doenças infecciosas responsáveis por causar prejuízos aos produtores rurais, levando a impactos econômicos significativos no agronegócio. Objetivou-se com o presente trabalho determinar o perfil epidemiológico da raiva em herbívoros no Estado de Pernambuco, Brasil, no período de 2007 a 2012. Foi realizado um estudo retrospectivo dos dados relativos aos casos positivos de raiva de herbívoros, levando em consideração o mês e o ano da ocorrência e a região geográfica. As análises moleculares foram desenvolvidas a partir de amostras de encéfalos provenientes das cinco Mesorregiões (Agreste, Mata, Sertão, Metropolitana e São Francisco) do Estado. No período estudado foram detectados 238 resultados positivos para o vírus da raiva em herbívoros, distribuídas espacialmente nas cinco mesorregiões, em 78 (42,1%) dos 185 municípios. Observou-se no decorrer do período uma diminuição significativa na taxa de incidência, com ausência de sazonalidade. Quando se analisou a taxa de incidência levando em consideração as Mesorregiões, observou-se que a Mata foi a que apresentou maior oscilação. Para complementar a análise epidemiológica, 16 amostras foram submetidas à técnica de RT-PCR para amplificação parcial do gene N. As sequências geradas foram alinhadas com sequências homólogas obtidas no GenBank para a construção da árvore filogenética, pelo método Bayesiana. Todas as amostras foram homólogas... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Rabies is an anthropozoonosis with lethal evolution caused by genus lyssavirus viruses. It is one among infectious diseases who are responsible for causing losses to farmers, leading to a significant economic impacts on agribusiness. The objective of this study was to determine the epidemiological profile in herbivores whithin the State of Pernambuco, Brazil, from 2007 to 2012. A retrospective study was conducted based on data from positive cases of rabies of herbivores, considering the month, the year of occurrence and geographic region. The molecular analyzes were developed using brain samples from the five Mesoregions (Agreste, Mata, Sertão, Metropolitana and São Francisco) within the State. During the study period, were detected positive for rabies 238 from herbivores, spatially distributed in 78 (42.1%) of 185 municipalites from the five mesoregions. It was observed, during the period cited, a significant decrease in the incidence rate, with no seasonal nature. While analyzing the incidence rate considering the mesoregions, the Mata region showed the greatest oscillation. As a complement to the epidemiological analysis, 16 samples were subjected to RT-PCR for the partial amplification of the gene N. The generated sequences were aligned with homologous sequences obtained from the GenBank to build the phylogenetic tree by bayesian method. On the results, it was found that the rabies virus is present all over the state of Pernambuco, related to the lineage of vampire bat De... (Complete abstract click electronic access below) / Doutor

One of the key events in a virus-infected host cell is the activation and repression of a large number of host genes. In recent years, such differentially expressed host genes have been identified for several viruses, bacteria and parasites. Such studies indicate that reprogramming of host transcriptome during infection by a pathogen is a major component of host response and many of the reprogrammed genes may promote or prevent pathogen infection or may contribute to pathogen-induced pathological changes. Host gene expression changes have been studied for a number of viruses in cell lines. However, in case of neurotropic viruses which infect nonrenewable populations of central nervous system (CNS), changes in the host gene expression need to be studied in the intact host rather than cells grown in culture. Since such studies are reported only for a few neurotropic viruses, an attempt has been made in this thesis to identify and characterize genes that are differentially expressed in the mouse brain during infection by Japanese encephalitis virus (JEV) and rabies virus. Using subtraction hybridization technique, subtraction cDNA libraries were generated representing mRNAs that are induced or repressed in the mouse brain during JEV infection. Sequence analysis of the 350 isolated clones resulted in the identification of 73 unique genes. Out of these, 66 were of forward library clones (upregulated genes) and 7 of reverse library clones (downregulated genes). The forward library clones was clustered in different functional groups such as, proteins involved in immune response and interferon-inducible proteins, GTPase and GTP binding proteins, transcriptional regulators, enzymes, ribosomal proteins, neuronal proteins, carrier proteins, DNA-binding proteins, miscellaneous and proteins of unknown function. The differential expression of all these genes was further validated by northern blot analysis of brain RNA isolated from normal and JEV-infected mice, which indicate that out of 66 forward library clones 33 were genuinely upregulated in JEV-infected mouse brain, whereas all 7 reverse library clones were repressed in JEV infection. Since vaccination is known to prevent virus replication in the brain, host gene expression changes in mice immunized with BIKEN JE vaccine was also examined. There was a good correlation between inhibition of JEV replication and reduced expression of JEV-inducible CNS genes in the vaccinated mice. To check whether JEV-induced CNS genes identified in this study are specific to JEV or can be induced by any other neurotropic virus, expression patterns of 15 randomly chosen genes were checked in RV infected mouse brain. Results indicated that all the chosen genes are modulated in the same way during RV infection as well. Comparison of JEV-induced gene expression changes with those induced by other neurotropic viruses indicated that 83% of the JEV-inducible mouse CNS genes are also induced by Sindbis virus, a neurotropic virus of the family alphaviridae, indicating that despite diverse life cycles, these two viruses may activate common host signaling pathways. This study also led to the identification of 9 unique JEV-inducible genes (LRG-21, VHSV induced gene1, Tpt1, SLC25A3, Olfm1, Ina/NF-66, Dst/Bpag1, Mdm2 and Gbp5) which are not reported to be activated by any other neurotropic virus. Since it is beyond the scope of this study to characterize all the JEV-induced and repressed genes, two genes were chosen for a detailed analysis. These are: JEV-inducible gene encoding GARG-39 protein which is a member of the glucocorticoid attenuated response gene family and an unannotated, JEV-repressible gene designated in this study as clone # 137. The gene encoding GARG-39 identified as a JEV-inducible gene in this study was originally discovered as lipopolysaccharide- and interferon-inducible gene in macrophages. This protein contains tetratricopeptide repeat (TPR) motifs that are known to be involved in protein-protein interactions. However, the function of this protein remains unknown till date. Therefore the gene was cloned and over-expressed in E. coli and antibodies were raised against the recombinant protein. Western blot analysis revealed that GARG-39 protein is detectable only in JEV-infected but not in the normal mouse brain. Surprisingly, immunoflourescence studies carried out in NIH3T3 cells revealed that GARG-39 is localized in the cytosol of normal cells and it colocalizes with α-tubulin in the mitotic spindle in a small fraction of cells which are in the mitotic stage. Further, in an in vitro assay, GARG-39 was found to interact with taxol-stabilized tubulin polymers. Since microtubules are known to play an important role in virus assembly, it is possible that GARG-39 may have a role in virus assembly and maturation. Alternatively, microtubule-associated proteins are implicated in several neurodegenerative disorders including Parkinson’s, Alzheimer’s and mental retardation and therefore, a role for GARG-39 in virus-induced neuropathogenesis cannot be ruled out. In addition, the expression of GARG-39 in normal dividing cells in the culture indicates a role for this protein in mitosis. In a normal mouse brain, mitotically active cells are very low in number and hence GARG-39 expression (both at the RNA and protein levels) is below the detection limits. JEV infection may trigger mitotic activity in brain leading to increased expression of GARG-39. One of the cDNA clones identified in this study, designated as clone # 137, hybridized to a ~2.6 kb transcript which was found to be down regulated in the mouse brain by JEV as well as rabies virus. A series of investigations led to the conclusion that clone #137 corresponds to the 3′ end of a ~2.6 kb transcript encoding mouse calcium calmodulin kinase inhibitor II α (mCaMKIINα). Interestingly, only the α isoform but not the β isoform of mCaMKIINα mRNA is down regulated in the mouse brain during JEV infection. Since the physiological function of mCaMKIINα is not known, the gene encoding 8 kDa mouse mCaMKIINα open reading frame was cloned into an E. coli expression vector and antibodies were raised against the purified recombinant protein. Surprisingly, antibodies raised against the ~8 kDa recombinant mouse CaMKIINα reacted with a ∼37 kDa mouse brain protein. This protein designated as CaMKIINα-immunoreactive protein (CaMKIINα-IRP) is also down regulated during JEV infection and is localized in the post synaptic density (PSD) of normal mouse brain. In addition, distinct changes are also observed in the subcellular localization and phosphorylation of CaMKIIα leading to an increase in cytosolic CaMKII activity in JEV-infected mouse brain. The differential regulation of CaMKIIα and CaMKIINα during JEV infection suggests a possible role for CaMKII signaling pathway in JEV infection and/or JEV-induced neuropathogenesis in the CNS. Conclusions: • A number of host genes whose expression is modulated in the mouse brain during JEV and/or rabies virus infection have been identified. • One of the JEV-inducible genes encoding the GARG-39 protein was shown to be a microtubule-associated protein with a possible role in mitosis. • One of the JEV-repressible genes was found to encode the mouse CaMKIINα mRNA. • A novel JEV-repressible ∼37 kDa protein immunoreactive to antibodies raised against the recombinant CaMKIINα was identified in the post synaptic density of the mouse brain.

Pasiutligė – viena iš seniausių ir pavojingiausių žmonių ir gyvūnų ligų. Šia liga užsikrečiama kai pasiutlige sergantis gyvūnas įkanda žmogui ar kitam gyvūnui, ar apseilėja sužalotą odą. Žmonės dažniausiai užsikrečia nuo valkataujančių naminių gyvūnų, todėl išlieka visiems reali užsikrėtimo pasiutligės virusu grėsmė. Vienintelis ir efektyviausias apsisaugojimo būdas nuo pasiutligės yra imunoprofilaktika. Šiame darbe ištirtas pasiutligės viruso paplitimas 2003-2011 metų laikotarpiu ir nustatyta laukinių ir naminių gyvūnų pasiutligės epidemiologinė situacija Lietuvoje. Atliekant 2007–2011 metų pasiutligės geografinio paplitimo analizę buvo nustatyta, kad nagrinėjamu laikotarpiu didžiausias tiek laukinių, tiek naminių gyvūnų infekuotumas buvo nustatytas Lietuvos Rytinėje ir Pietrytinėje dalyje. Naudoti pasiutligės diagnostikos metodai pasižymi speciškumu ir jautrumu. Įrodyta, kad kartu taikant tiesioginį fluorescuojančių antikūnų ir audinių kultūrų infekavimo pasiutligės virusu metodus, galima užtikrinti greitą ir efektyvų pasiutligės viruso nustatymą laukinių ir naminių gyvūnų tiriamuosiuose galvos smegenų mėginiuose, tuo užtikrinant diagnozės patvirtinimą dėl užsikrėtimo pasiutligės virusu. Laukinių gyvūnų oralinės vakcinacijos nuo pasiutligės (ORV) periodu 2007-2011 metais atlikta naminių ir laukinių gyvūnų pasiutligės viruso izoliatų filogenetinė analizė N geno srityje atvirkštinės transkripcijos polimerazės grandininės reakcijos (AT-PGR) metodu. Pirmą... [toliau žr. visą tekstą] / Rabies is one of the oldest and most dangerous human and animal diseases. This is a disease that is transmitted directly from animal to animal and from an animal to a human being. People usually contract this disease from stray domestic animals therefore a realistic threat is posed to everyone to become infected with the rabies virus (RV). The only and the most effective way of protecting oneself from rabies after an infected or unknown animal has bitten one, is immunoprophylaxis. Between 2003 and 2011, the epidemiological situation of rabies in Lithuania was assessed. The methods that were used to diagnose rabies are noted for specificity and sensitivity. It has been proved that applying simultaneously the direct fluorescent antibody test (FAT) and rabies tissue culture infections test (RTCIT) methods it was possible to ensure a fast and effective determination of RV in the samples of wild and domestic animals under investigation thereby ensuring confirmation of the diagnosis of being infected with RV. During the wild fauna oral rabies vaccination (ORV) period between 2007 and 2011, the philogenetic analysis of RV isolates of domestic and wild animals in the sphere of N gene by means of the reverse transcription-polymerase chain reaction (RT-PCR) was carried out for the first time. In 2006-2011, the assessment of the efficacy of ORV in raccoon dogs and red foxes by means of the quantitative the Enzyme-linked immunoassay test (ELISA) research method... [to full text]

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Rabies is one of the oldest and most dangerous human and animal diseases. This is a disease that is transmitted directly from animal to animal and from an animal to a human being. People usually contract this disease from stray domestic animals therefore a realistic threat is posed to everyone to become infected with the rabies virus (RV). The only and the most effective way of protecting oneself from rabies after an infected or unknown animal has bitten one, is immunoprophylaxis. Between 2003 and 2011, the epidemiological situation of rabies in Lithuania was assessed. The methods that were used to diagnose rabies are noted for specificity and sensitivity. It has been proved that applying simultaneously the direct fluorescent antibody test (FAT) and rabies tissue culture infections test (RTCIT) methods it was possible to ensure a fast and effective determination of RV in the samples of wild and domestic animals under investigation thereby ensuring confirmation of the diagnosis of being infected with RV. During the wild fauna oral rabies vaccination (ORV) period between 2007 and 2011, the philogenetic analysis of RV isolates of domestic and wild animals in the sphere of N gene by means of the reverse transcription-polymerase chain reaction (RT-PCR) was carried out for the first time. In 2006-2011, the assessment of the efficacy of ORV in raccoon dogs and red foxes by means of the quantitative the Enzyme-linked immunoassay test (ELISA) research method... [to full text] / Pasiutligė – viena iš seniausių ir pavojingiausių žmonių ir gyvūnų ligų. Šia liga užsikrečiama kai pasiutlige sergantis gyvūnas įkanda žmogui ar kitam gyvūnui, ar apseilėja sužalotą odą. Žmonės dažniausiai užsikrečia nuo valkataujančių naminių gyvūnų, todėl išlieka visiems reali užsikrėtimo pasiutligės virusu grėsmė. Vienintelis ir efektyviausias apsisaugojimo būdas nuo pasiutligės yra imunoprofilaktika. Šiame darbe ištirtas pasiutligės viruso paplitimas 2003-2011 metų laikotarpiu ir nustatyta laukinių ir naminių gyvūnų pasiutligės epidemiologinė situacija Lietuvoje. Atliekant 2007–2011 metų pasiutligės geografinio paplitimo analizę buvo nustatyta, kad nagrinėjamu laikotarpiu didžiausias tiek laukinių, tiek naminių gyvūnų infekuotumas buvo nustatytas Lietuvos Rytinėje ir Pietrytinėje dalyje. Naudoti pasiutligės diagnostikos metodai pasižymi speciškumu ir jautrumu. Įrodyta, kad kartu taikant tiesioginį fluorescuojančių antikūnų ir audinių kultūrų infekavimo pasiutligės virusu metodus, galima užtikrinti greitą ir efektyvų pasiutligės viruso nustatymą laukinių ir naminių gyvūnų tiriamuosiuose galvos smegenų mėginiuose, tuo užtikrinant diagnozės patvirtinimą dėl užsikrėtimo pasiutligės virusu. Laukinių gyvūnų oralinės vakcinacijos nuo pasiutligės (ORV) periodu 2007-2011 metais atlikta naminių ir laukinių gyvūnų pasiutligės viruso izoliatų filogenetinė analizė N geno srityje atvirkštinės transkripcijos polimerazės grandininės reakcijos (AT-PGR) metodu. Pirmą... [toliau žr. visą tekstą]

In Chapter 1, the literature relating to rabies virus and the rabies like lyssaviruses is reviewed. In Chapter 2 data are presented from 1170 diagnostic submissions for ABLV testing by fluorescent antibody test (Centocor FAT). All 27 non-bat submissions were ABLV-negative. Of 1143 bat accessions 74 (16%) were ABLV-positive, including 69 of 974 (7.1%) flying foxes (Pteropus spp.), 5 of 7 (71.4%) Saccolaimus flaviventris (Yellow-bellied sheathtail bats), none of 151 other microchiropteran bats, and none of 11 unidentified bats. Statistical analysis of data from 868 wild Black, Grey-headed, Little Red and Spectacled flying foxes (Pteropus alecto, P. poliocephalus, P. scapulatus, and P. conspicillatus) indicated that three factors; species, health status and age were associated with significant (p&lt 0.001) differences in the proportion of ABLV-positive bats. Other factors including sex, whether the bat bit a person or animal, region, year, and season submitted, were not associated with ABLV. Case data for 74 ABLV-positive bats, including the circumstances in which they were found and clinical signs, is presented. In Chapter 3, the aetiological diagnosis was investigated for 100 consecutive flying fox submissions with neurological signs. ABLV (32%), spinal and head injuries (29%), and neuro-angiostrongylosis (18%) accounted for most neurological syndromes in flying foxes. No evidence of lead poisoning was found in unwell (n=16) or healthy flying foxes (n=50). No diagnosis was reached for 16 cases, all of which were negative for ABLV by TaqMan® PCR. The molecular diversity of ABLV was examined in Chapter 4 by sequencing 36 bases of the leader sequence, the entire N gene, and start of the P gene of 28 isolates from pteropid bats and 3 isolates from Yellow-bellied sheathtail (YBST) bats. Phylogenetic analysis indicated all ABLV isolates clustered together as a discrete group within the Lyssavirus genera closely related to rabies virus and European bat lyssavirus-2 isolates. The ABLV lineage consisted of two variants; one (ybst-ABLV) consisted of isolates only from YBST bats, the other (pteropid-ABLV) was common to Black, Grey-headed and Little Red flying foxes. No associations were found between the sequences and either the geographical location or year found, or individual flying fox species. In Chapter 5, 15 inocula prepared from the brains or salivary glands of naturally-infected bats were evaluated by intracerebral (IC) and footpad (FP) inoculation of Quackenbush mice in order to select and characterize a highly virulent inoculum for further use in bats (Inoculum 5). In Chapter 6, nine Grey-headed flying foxes were inoculated with 105.2 to 105.5 MICED50 of Inoculum 5 divided into four sites, left footpad, pectoral muscle, temporal muscle and muzzle. Another bat was inoculated with half this dose divided into the footpad and pectoral muscle only. Seven of 10 bats developed clinical disease of 1 to 4 days duration between PI-days 10 and 19 and were shown to be ABL-positive by FAT, HAM immunoperoxidase staining, virus isolation in v mice, and TaqMan PCR. Five of the seven bats displayed overt aggression, one died during a seizure, and one showed intractable agitation, pacing, tremors, and ataxia. Viral antigen was demonstrated throughout the central and peripheral nervous systems and in the epithelial cells of the submandibular salivary glands (n=4). All affected bats had mild to moderate non-suppurative meningoencephalitis and severe ganglioneuritis. No ABLV was detected in three bats that remained well until the end of the experiment on day 82. One survivor developed a strong but transient antibody response. In Chapter 7, the relative virulence of inocula prepared from the brains and salivary glands of experimentally infected flying foxes was evaluated in mice by IC and FP inoculation and TaqMan assay. The effects in mice were correlated to the TaqMan CT value and indicated a crude association between virulence and CT value that has potential application in the selection of inocula. In Chapter 8, 36 Black and Grey-headed flying foxes were vaccinated with one (day 0) or two (+ day 28) doses of Nobivac rabies vaccine and co-vaccinated with keyhole limpet haemocyanin (KLH). All bats responded to the Nobivac vaccine with a rabies-RFFIT titer &gt 0.5 IU/mL that is nominally indicative of protective immunity. Plasma from bats with rabies titres &gt 2 IU/mL had cross-neutralising ABLV titres &gt 1:154. A specifically developed ELISA detected a strong but transient response to KLH.