This thesis deals with infectious diseases transmitted by arthropods in the Czech Republic in period of 2002 - 2011. Meeting the objectives and answer the research questions, I managed using secondary analysis of reported cases infectious diseases transmitted by arthropods in the information system for reporting and recording of infectious diseases - EPIDAT and literature review and subsequent analysis of scientific articles. Important additional informations were provided by State Veterinary Institute in Prague and the National Reference Laboratory for arboviruses in Ostrava. Thesis does not deal with number of single values, but it is focused on epidemiological severity of analyzed cases, severity of clinical course and risk of introducing infection in the Czech Republic. Theoretical part of the thesis describes selected arboviruses infections. Tick-borne encephalitis is one of the most important diseases transmitted by arthropods in our surroundings. Number of reported cases of tick-borne encephalitis was quite constant in monitored period 2002 - 2011, according to my research and statistical verification. Highest morbidity was in 2006. The same year the highest incidence of disease Tick-borne encephalitic was reported in region of Vysočina. This number is far closer to number of reported cases in region of South Bohemia which was the most affected region thought the entire period. Prevention exists against tick-borne encephalitis. It is vaccination which is recommended for people staying in natural foci of infection in the Czech Republic and abroad. In the Czech republic becomes more important West Nile fever. Increased incidence of this disease is recorded in the USA, but also in many European countries (eg Greece, Hungary, Italy) in recent years. One of the goals of this thesis is to characterize the surveillance of West Nile fever in the Czech republic. Monitoring antibody titer against West Nile virus in the serum of horses is one of the basic elements of surveillance of this disease in our country. 5 samples of the blood serum was increased titers of antibodies to West Nile virus. Various geographic distribution of horses and high specific antibody titers indicate increasing activity of West Nile virus. Clinical disease of horses not being recorded in the Czech republic. In the field of human medicine have been reported three imported cases of West Nile fever. Other arboviruses infections, that appeared in our country, is dengue disease and viral disease Chikungunya. In the monitored period there were a total of 98 reported cases of imported disease. Most cases were imported from India. Three cases of Chikungunya disease were reported in 2006 in connection with a stay on the island of Mauritius, where was the epidemic at this time. People can travel freely and there is the risk of importing diseases from the tropics and subtropics. Some infections transmitted by arthropods are in areas where it was previously absent recently. Prevention is essential. Specific and non-specific prevention. Another important element of prevention is strict adherence to methods for surveillance of disease.Viral infections transmitted by arthropods are among the most important emergent infectious diseases. In addition to malaria, malnutrition and helminth infection are the largest public health problem, not only in third world countries.

The thesis is devoted to the issue of vaccination problematics and awareness of tick-borne encephalitis in a selected population group in South Bohemia. For the needs of research, the selected group was composed of students of secondary schools and grammar schools in the South Bohemian Region in the age group from 13 to 19 years old. The objectives of this work was to analyze the incidence of tick-borne encephalitis in the Czech Republic and the South Bohemian Region between 2000 and 2015, to compare the vaccination in the Czech Republic with the other European countries, to find the vaccination of a selected age group in the South Bohemian Region and to analyze the level of awareness of the risks of tick-borne encephalitis by the polling method and the attitude of the age group to vaccination against this disease. To achieve the objectives was used a secondary analysis of the data from the EpiDat system, where the incidence of the disease was detected in the period in the South Bohemian Region and the Czech Republic. The questionnaire survey was used to obtain awareness on observed disease and attitude of the study group towards vaccination against tick-borne encephalitis. The conclusions of the questionnaire survey were obtained on the basis of statistical processing in the MS Excel spreadsheet. For my significant findings from my research, I consider 80% of the vaccination in the monitored group. Information on tick-borne encephalitis in most cases did not get the respondents from a doctor, but from family members, from the internet or from television. However, information on vaccination is given to respondents to a greater extent. Contributions by insurance companies do not motivate more than half of respondents to inoculation. Research has also shown, that respondents have insufficient information about the amount of the contribution and whether their insurance company provide the contribution at all. In view of these circumstances, I consider the increased promotion of this vaccination by insurance companies to be very desirable.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Canine distemper is one of the most prevalent viral diseases of dogs. Several cases are diagnosed in a daily basis in private practices and veterinary hospitals around the country. Most cases of neurological manifestation of canine distemper are fatal and the diagnosis confirmation by histopathology is often necessary. Canine distemper encephalitis is the main cause of death or reason for euthanasia in dogs necropsied at the Laboratório de Patologia Veterinária (LPV) of the Universidade Federal de Santa Maria (UFSM). Consequently the large number of canine distemper cases in the LPV-UFSM files prompted the performance of a retrospective study consisting of 620 neurologic cases of canine distemper which served as the basis for a prospective study on the neuro-histopathologic aspects of the disease. The current study was carried out to perform a detailed histopathologic investigation on the changes in the central nervous system (CNS) of dogs affect by canine distemper aiming to help students of veterinary pathology and veterinary pathologists in the correct diagnose of this disease. Seventy dogs necropsied at the LPV-UFSM and with diagnosis of canine distemper confirmed by the finding of characteristic inclusion bodies in the CNS were included in the study. In order to determine the prevalence of the lesions, several anatomical regions were selected from the brain and spinal cord; these sites were consistently microscopically examined in each case. Overall, most affected anatomical regions were, in decreasing order of frequency: cerebellum (91.4%), diencephalon (78.6%), frontal lobe (75.7%), pons (72.9%) and mesencephalon (70.0%). Demyelination was the most prevalent lesion; it was observed in 91.4% of the cases and was located mainly in the cerebellum (88.6%), pons (65.7%) and diencephalon (61.4%). The five structures most affected by demyelination were: roof of the fourth ventricle (68.6%), cerebellar folia (61.4%), cerebellar medulla (61.4%), cervical spinal cord (46.3%) and mesencephalic tegment (40.0%). Other lesions and their prevalence were non-suppurative encephalitis (70.0%), non-suppurative leptomeningitis (44.3%), non-suppurative myelitis (35.2%), encephalomalacia (31.4%), nonsuppurative myelitis (18.5%), laminar cortical necrosis (17.1%), myelomalacia (13.0%), nonsuppurative ependymitis (7.1%) and plexochoroiditis (1.4%). The occurrence of the inclusion bodies in the cells of the CNS was: astrocytes (82.8%), neurons (44.3%), gemistocytes (31.4%), ependymal cells (22.8%), meningothelial cells (4.3%) and cells of the choroids plexus (1.4%). / A cinomose é uma das doenças virais mais prevalentes em cães. Muitos cães são diagnosticados com a doença diariamente em clínicas e hospitais veterinários de todo o país. A forma neurológica da cinomose é fatal na grande maioria das vezes, e a confirmação do diagnóstico clínico através do exame histopatológico se faz necessária em muitos casos. A encefalite por cinomose representa a principal causa de morte ou razão para eutanásia de cães necropsiados no Laboratório de Patologia Veterinária (LPV) da Universidade Federal de Santa Maria (UFSM). Com isso, o grande número de casos de cinomose arquivados no LPVUFSM proporcionou a realização de um estudo retrospectivo de 620 casos da forma neurológica da doença que serviu como base para a realização de um estudo neurohistopatológico prospectivo. O presente estudo tem como objetivo realizar uma investigação histopatológica detalhada das alterações no sistema nervoso central (SNC) de cães com cinomose para auxiliar estudantes de patologia e patologistas no diagnóstico da doença. Para isso, foram selecionados 70 casos da forma neurológica da cinomose confirmada pelo achado dos corpúsculos de inclusão característicos no SNC em cães necropsiados no LPV-UFSM. Para determinar a prevalência das lesões, diversas regiões anatômicas do encéfalo e medula espinhal foram selecionadas e sistematicamente examinadas histologicamente em cada caso. As regiões anatômicas mais afetadas, em ordem decrescente, independentemente do tipo de lesão, foram: cerebelo (91,4%), diencéfalo (78,6%), lobo frontal (75,7%), ponte (72,9%) e mesencéfalo (70,0%). Desmielinização foi a lesão mais prevalente, observada em 91,4% dos casos, localizada principalmente no cerebelo (88,6%), ponte (65,7%) e diencéfalo (61,4%). As cinco estruturas anatômicas mais afetadas pela desmielinização foram: teto do quarto ventrículo (68,6%), folhas cerebelares (61,4%), substância medular do cerebelo (61,4%), medula espinhal cervical (46,3%) e tegmento mesencefálico (40,0%). A prevalência das outras lesões foi: encefalite não-supurativa (70,0%), leptomeningite não-supurativa (44,3%), mielite não-supurativa (35,2%), encefalomalacia (31,4%), meningomielite não-supurativa (18,5%), necrose laminar cortical (17,1%), mielomalacia (13,0%), ependimite não-supurativa (7,1%) e plexocoroidite não-supurativa (1,4%). A ocorrência dos corpúsculos de inclusão nas células do SNC foi: astrócitos (82,8%), neurônios (44,3%), gemistócitos (31,4%), células do epêndima (22,8%), células meningoteliais (4,3%) e células do plexo coróide (1,4%).

Submitted by Simone Souza (simonecgsouza@hotmail.com) on 2017-09-15T13:37:53Z No. of bitstreams: 1 DISS_2014_Letícia Borges da Silva Heinen.pdf: 4369897 bytes, checksum: 2e00a8b07d093787dbdb2e23dd6ac34e (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2017-09-19T12:59:17Z (GMT) No. of bitstreams: 1 DISS_2014_Letícia Borges da Silva Heinen.pdf: 4369897 bytes, checksum: 2e00a8b07d093787dbdb2e23dd6ac34e (MD5) / Made available in DSpace on 2017-09-19T12:59:17Z (GMT). No. of bitstreams: 1 DISS_2014_Letícia Borges da Silva Heinen.pdf: 4369897 bytes, checksum: 2e00a8b07d093787dbdb2e23dd6ac34e (MD5) Previous issue date: 2014-03-28 / CNPq / O gênero Flavivirus, família Flaviviridae, alberga arbovírus como os vírus dengue (DENV) e da febre amarela, que possuem importância médica e são envolvidos em epidemias de doença febril em áreas urbanas e rurais em regiões tropicais e subtropicais. No Brasil, atualmente, o DENV e o vírus da encefalite de Saint Louis (SLEV) são os dois flavivírus circulantes em áreas urbanas mais frequentes. Desde a introdução e emergência dos diferentes sorotipos de DENV a partir da década de 1980, extensas epidemias de febre do dengue vêm sendo reladas por todo o país. O SLEV, anteriormente reconhecido apenas em ciclos enzoóticos e com pouca relevância médica no Brasil, tem sido implicado em casos de doença febril durante epidemia de dengue no sudeste do país. O objetivo deste estudo foi investigar a circulação de flavivírus em pacientes com doença febril aguda com suspeita de dengue em Mato Grosso (MT) em 2011 e 2012. Material e Métodos: 604 amostras de soro obtidas entre outubro de 2011 e julho de 2012 de pacientes com doença febril aguda suspeita de dengue com até cinco dias do início dos sintomas em MT foram submetidas à multiplex semi-nested RT-PCR para a pesquisa de flavivírus, como os quatro sorotipos de DENV, SLEV, vírus da febre amarela, do Oeste do Nilo, Rocio, Bussuquara, Iguape e Ilhéus. Amostras positivas foram testadas em pelo menos duas reações independentes de single-nested RT-PCR e submetidas a sequenciamento nucleotídico de região do gene da glicoproteína de envelope para análise filogenética. Resultados: Dentre os 604 pacientes, 315 (52,2 %) foram positivos para DENV-4, 24 (4,0 %) para DENV-1, 3 (0,5 %) para SLEV, 1 (0,2 %) para DENV-2 e 1 (0,2 %) para DENV-3. Todas as amostras eram de pacientes oriundos de áreas urbanas de 17 municípios de MT. Entre as amostras positivas, 9 eram co-infecções entre DENV-1/DENV-4, 1 entre DENV-2/DENV-4, 2 por SLEV/DENV-4 e 1 entre SLEV/DENV-1/DENV-4. Os demais flavivírus pesquisados não foram detectados. Amostras negativas para flavivírus totalizaram 273/604 (45,20 %). Discussão: A ocorrência das arboviroses na população geralmente é subestimada, devido a fatores como quadro clínico inespecífico, infecções inaparentes e ausência de diagnóstico diferencial. O DENV-4 foi introduzido no MT em 2012, responsável pela maior casuística nas cidades da Baixada Cuiabana. Co-infecções são frequentes quando há circulação hiperendêmica dos quatro sorotipos do DENV, situação já relatada no Brasil em Manaus e Rio de Janeiro em 2011. O SLEV foi detectado em pacientes de Cuiabá e Várzea Grande. Infecções por SLEV são primariamente inaparentes ou brandas. Em MT, espécies de Culex e outros vetores deste virus são amplamente dispersas. Como humanos são hospedeiros finais e apresentam baixa viremia, sua ocorrência é provavelmente subestimada. Conclusão: DENV-1 e DENV-4 foram os flavivírus identificados com maior frequência. Os quatro sorotipos do DENV foram detectados em Cuiabá e infecções esporádicas pelo SLEV foram identificadas em pacientes co-infectados com o DENV-4 ou o DENV-4/DENV-1 em Cuiabá e Várzea Grande, indicando que outros arbovírus podem circular silenciosamente durante epidemia de dengue em áreas urbanas em MT. / The genus Flavivirus, Flaviviridae family, comprises arboviruses such as the medical important dengue virus (DENV) and yellow fever virus, involved in febrile illness epidemics in urban and rural areas of tropical and subtropical regions. In Brazil, DENV and Saint Louis encephalitis (SLEV) virus are currently the two most important flaviviruses circulating in urban areas. Since the introduction and emergence of different DENV serotypes in the 1980´s, extensive dengue outbreaks have been reported throughout the country.SLEV, previously recognized only in enzootic cycles without medical relevance in Brazil, has been implicated to febrile illness etiology during dengue fever outbreaks in the Southeast region. The aim of this study was to investigate the circulation of flaviviruses in patients with acute febrile illness suspected of harboring dengue in Mato Grosso (MT) between 2011 and 2012. Material and Methods: 604 serum samples obtained between October 2011 and July 2012 from patients with acute febrile illness suspected of dengue lasting less than 5 days in MT were subjected to multiplex semi-nested RT-PCR for flaviviruses, including all four serotypes of DENV, SLEV Yellow Fever, West Nile, Rocio, Bussuquara, Iguape and Ilheus viruses. Positive samples were tested at least twice in independent single-nested RT-PCR reactions and subjected to nucleotide sequencing of the envelope glycoprotein (E) gene region for phylogenetic analysis. Results: Among 604 patients, 315 (52.2 %) were positive for DENV-4, 24 (4.0 %) for DENV-1, three (0.5 %) for SLEV, one (0.2 %) for DENV-2 and one (0.2 %) for DENV-3. All patients are residents in urban areas of 17 cities of MT. Among then, 9 were co-infections among DENV-1/DENV-4, 1 between DENV-2/DENV-4, two between SLEV/DENV-4 and one with SLEV/DENV-1/DENV-4. The other flaviviruses were not detected. Negative samples for flavivirus totaled 273/604 (45.20 %). Discussion: The occurrence of arboviruses in the population generally is underestimated, probably due to unapparent infection or unspecific clinical presentation, associated to the absence of differential diagnosis. The DENV-4 serotype was introduced in MT in 2012, responsible for the largest number of cases in Cuiabá and Varzea Grande. Co-infections are common when hiperendemic circulation of all four serotypes of DENV is observed. This situation has already been reported in Brazil in Manaus and Rio de Janeiro cities in 2011. Three patients were positive for SLEV in Cuiaba and Várzea Grande. SLEV infections are primarily mild or unapparent. In MT, species of Culex and other vectors are widely dispersed. As humans are final hosts and, therefore, present low titer viremia, the occurrence of SLEV in the population is probably underestimated. Conclusion: DENV-1 and DENV-4 were the most frequently flaviviruses identified. The four DENV serotypes were detected in Cuiaba and sporadic SLEV infections were identified in patients co-infected with DENV-4 or DENV-1/DENV-4 in Cuiaba and Várzea Grande, indicating that other arboviruses may circulate silently during dengue epidemics in urban areas of MT.

Submitted by Valquíria Barbieri (kikibarbi@hotmail.com) on 2018-04-20T17:14:55Z No. of bitstreams: 1 DISS_2015_Otacília Pereira Serra.pdf: 1299760 bytes, checksum: adf22b742c77d45b935dd95c6e46cbda (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2018-04-27T17:52:07Z (GMT) No. of bitstreams: 1 DISS_2015_Otacília Pereira Serra.pdf: 1299760 bytes, checksum: adf22b742c77d45b935dd95c6e46cbda (MD5) / Made available in DSpace on 2018-04-27T17:52:07Z (GMT). No. of bitstreams: 1 DISS_2015_Otacília Pereira Serra.pdf: 1299760 bytes, checksum: adf22b742c77d45b935dd95c6e46cbda (MD5) Previous issue date: 2015-04-20 / CAPES / FAPEMAT / Arbovírus são transmitidos por artrópodes hematófagos, representando um problema de saúde pública em áreas tropicais. O objetivo deste estudo foi investigar a diversidade de espécies de culicídeos e sua frequência de infecção por Alphavirus e Flavivirus em Cuiabá, MT. Foram realizadas capturas com aspirador de Nasci e puçá entre janeiro e abril de 2013 em três locais de 200 setores censitários, definidos aleatoriamente. Os culicídeos foram identificados com chave dicotômica de Forattini, alocados em pools (1-20 mosquitos) segundo sexo, espécie, data, local de coleta e armazenados a -80˚C. Pools de fêmeas foram submetidos à extração de RNA e DNA total, à multiplex semi-nested-RT-PCR para cinco alphavírus e 11 flavivírus e Nested- PCR para identificação de Culex (Cx.) quinquefasciatus. Amostras positivas para SLEV, DENV-1, -4 e MAYV foram submetidas a single RT-PCR e sequenciamento nucleotídico. Pools positivos para o MAYV foram inoculados em células Vero e submetidos a RT-PCR para o gene de envelope E1 dos alphavirus. Pools positivos para flavivirus foram inoculados em células C6/36 (Flavivirus). Calculou-se a taxa de infecção mínima (MIR). Foram capturados 11.090 mosquitos, 4.556 fêmeas de 14 espécies, perfazendo 610 pools. Foram analisados 171 pools de Aedes (Ae.) aegypti; 1 Ae. albopictus; 1 Aedes sp.; 5 Cx. bidens/interfor; 1 Cx. spinosus; 403 Cx. quinquefasciatus; 1 Galindomyia sp; 6 Limatus sp; 2 Mansonia wilsoni; 5 Psorophora (Ps.) sp; 1 Ps. ciliata; 11 Ps. varipes/albigenu; 1 Sabethes chloropterus e 1 Uranotaenia sp. Encontrou-se 1/171 (MIR=0,92) pool de Ae. aegypti positivo para DENV-1; 1/403 (MIR= 0,2) Cx. quinquefasciatus para SLEV genótipo V-A, 12/403 (MIR=3,5) de Cx. quinquefasciatus, 4/171 (MIR=3,67) de Ae. aegypti para MAYV; destes, cinco pools apresentaram co-infecção com DENV-4. Um produto de 1,3 kb obtido com o protocolo para o gene de envelope de três pools positivos para o MAYV resultou em sequências nucleotidicas inespecíficas. O MAYV foi isolado de dois pools contendo duas fêmeas não ingurgitadas de Ae. aegypti (#958) e duas de Cx. quinquefasciatus (#489). Positividade para DENV-4 foi identificada em 58/171 (MIR=53,35) Ae. aegytpi, 105/403 (MIR=30,65) Cx. quinquefasciatus, 2/5 (MIR=400) Psorophora sp, 2/11 (MIR=142,85) Ps. varipes/albigenu, 1/1 (MIR= 1000) Sabethes chloropterus, 2/5 (MIR=285.7) Cx. bidens/interfor e 1/1 (MIR=1000) Aedes sp. O DENV-4 foi isolado de dois pools contendo três (#329) e 16 (#806) fêmeas de Cx. quinquefasciatus não ingurgitadas. O SLEV, MAYV e os sorotipos do DENV foram identificados em pacientes com suspeita de dengue na cidade de Cuiabá em estudos prévios do Laboratório de Virologia. Experimentalmente, Culex e Aedes spp. são vetores competentes do MAYV. A identificação do vírus em fêmeas não ingurgitadas sugere que estas espécies podem estar envolvidas no ciclo urbano do MAYV em Cuiabá. Dentre os sorotipos do DENV, somente o DENV-1 e o DENV-4 foram identificados em culicídeos. O DENV-4 tem produzido importantes epidemias em MT desde 2012. A positividade para DENV-4 em diferentes espécies de culicídeos pode ser decorrente de infecção natural ou da hematofagia em humanos, pois muitos destes pools apresentavam fêmeas ingurgitadas. Cuiabá possui ecossistema favorável para a ocorrência de arboviroses e proliferação de vetores. Estudos envolvendo vigilância entomológica e virológica são importantes para estimar a situação epidemiológica dos arbovírus no estado. / Arbovirus are transmitted by hematophagous arthropods, posing a public health issue in tropical areas. The aim of this study was to investigate the diversity of culicidae and their frequency of infection by arboviruses from Alphavirus and Flavivirus genus in Cuiabá, MT. To achieve that, culicids were captured with Nasci aspirators and hand net between January and April 2013 in three locations of 200 censitary sectors randomly defined. The specimens were identified using the Forattini dichotomy key, allocated in pools (1-20 mosquitoes), according to sex, species, day and place of capture, and stored at -80˚C. Female pools were subjected to total RNA and DNA extraction and to multiplex semi-nested-RT-PCR for five alphaviruses and 11 flaviviruses and Nested-PCR for Culex (Cx.) quinquefasciatus identification. The pools positive for SLEV, DENV-1, -4 and MAYV were subjected to single RT-PCR and nucleotide sequencing. MAYVpositive pools were inoculated in Vero cells and subjected to RT-PCR for the E1 envelope gene of alphaviruses. Pools positive for flaviviruses were inoculated in C6/36 cells. The minimum infection rate (MIR) was calculated. 11,090 mosquitoes were captured, 4,556 females belonging to 14 species, comprising 610 pools, 171 pools of Aedes (Ae.) aegypti specimens; 1 Ae. albopictus; 1 Aedes sp.; 5 Cx. bidens/interfor; 1 Cx. spinosus; 403 Cx. quinquefasciatus; 1 Galindomyia sp.; 6 Limatus sp.; 2 Mansonia wilsoni; 5 Psorophora sp.; 1 Ps. ciliata; 11 Ps. varipes/albigenu; 1 Sabethes chloropterus e 1 Uranotaenia sp. Among them, 1/171 (MIR=0.92) Ae. aegypti pool was positive for DENV-1 and 1/403 (MIR=0.3) Cx. quinquefasciatus for SLEV genotype V-A. For MAYV, 12/403 (MIR=3.5) Cx. quinquefasciatus, 4/171 (MIR=3.67) Ae. aegypti were positive, five of them were also infected by DENV-4. A DNA product with 1.3 kb obtained from three pools positive for MAYV with the protocol for the envelope gene resulted in unspecific nucleotide sequences. MAYV was isolated from two pools, both containing two non-engorged females of Ae. aegypti (#958) and Cx. quinquefasciatus (#489). DENV-4 was detected in 58/171 (MIR=53.35) Ae. aegytpi, 105/403 (MIR=30,65) Cx. quinquefasciatus, 2/5 (MIR=400) Psorophora sp., 2/11 (MIR=142.85) Ps. varipes/albigenu, 1/1 (MIR= 1000) Sabethes chloropterus, 2/5 (MIR=285.7) Cx. bidens/interfor and 1/1 (MIR=1000) Aedes sp. DENV-4 was isolated from two pools containing three (#329) and 16 (#806) non-engorged females of Cx. quinquefasciatus. The SLEV, MAYV and the four DENV serotypes were identified in patients suspected of harboring dengue infection in Cuiabá in previous studies of the Virology Laboratory. Experimentally, Culex and Aedes spp. are competent vectors for MAYV. The identification of the virus in nonengorged females suggests these species may be involved in the urban cycle of MAYV in Cuiabá. Among the DENV serotypes, only DENV-1 and DENV-4 were identified in culicids captured in the city. DENV-4 has been responsible for major outbreaks in MT since 2012. The identification of DENV-4 in several mosquito species might be resultant either from natural infection or hematophagy in humans, since several of these pools presented engorged females. Cuiabá presents a favorable ecosystem for the occurrence of arboviruses and vector proliferation. Studies involving entomological and virological surveillance are important to estimate the epidemiological situation of arboviruses in the state.

A saliva de insetos hematófagos possui moléculas capazes de modular o sistema imune do hospedeiro. Com base na literatura a respeito das atividades presentes na saliva de Aedes aegypti, investigamos se o EGS dessa espécie era capaz de modular a EAE. Imunizamos animais C57BL/6 com MOG35-55, e realizamos um tratamento com EGS. O tratamento com EGS diminuiu a incidência da doença e provocou um atraso no aparecimento dos sinais clínicos, além de estes serem mais brandos. Observamos que a modulação se deu na fase de indução da resposta imune, não na efetora. De fato, o EGS consegue suprimir a doença por 4 vias: 1) diminuindo a expressão de MHCII, CD80 e CD86 em células dendríticas, e diminuindo a produção de citocinas responsáveis pela indução das respostas Th1/Th17; 2) induzindo células produtoras de IL-10 in vivo; 3) induzindo apoptose em linfócitos T naive; 4) induzindo células com perfil Th2 produtoras de IL-4 e IL-5. Concluímos que o EGS é capaz de atuar na supressão dos sintomas durante o curso da EAE e na inibição do início da resposta imune. / The saliva of hematophagous insects has molecules that can modulate the host immune system. Based on the literature about activities found in Aedes aegypti saliva, we investigate if SGE of this species could modulate EAE. We have immunized C57BL/6 mice with MOG35-55, and carried out a treatment with SGE. The treatment with SGE reduced the incidence of disease and caused a delay onset of clinical signs making them softer. We have observed that modulation occured in the induction phase of immune response, not in effector phase. In fact, SGE can suppress the disease by four ways: 1) decreasing the expression of MHCII, CD80 and CD86 in dendritic cells and decreasing the production of cytokines responsible for Th1/Th17 response induction; 2) inducing cells producing IL-10 in vivo; 3) inducing apopotosis in naive T lymphocytes; 4) inducing cells Th2 producing IL-4 e IL-5. We came to the conclusion that SGE can act in supressing symptoms during the course of EAE and inhibiting the beggining of autoimmune response.

Infecções no sistema nervoso central (SNC) causadas por microrganismos desencadeiam sintomas de moderados a severos, dependendo da região atingida, podendo ser designadas como encefalites ou meningites. Os vírus são os agentes mais comuns nestas infecções. Os agentes virais responsáveis por essas enfermidades que apresentam maior incidência na população mundial são certos herpesvírus, flavivírus, influenza A, enterovírus e vírus da caxumba. Entretanto, essa prevalência varia de acordo com a população, estado imunológico do indivíduo, idade e região estudada. Embora existam dados bem estabelecidos da etiologia dessas doenças em alguns países, ainda há uma carência de informação no que diz respeito à etiologia dessas moléstias no Brasil. Assim, informações mais precisas em relação à prevalência desses agentes em nosso meio são necessárias para o desenvolvimento e aplicação de métodos de diagnósticos mais rápidos e eficientes. Neste trabalho, foram analisadas 120 amostras de liquido cefalorraquidiano (LCR), procedentes de dois centros da cidade de São Paulo (Irmandade Santa Casa de Misericórdia e Hospital das Clínicas da Faculdade de medicina da Universidade de São Paulo), as quais foram submetidas à reação em cadeia de polimerase para o herpesvirus simples 1 e 2 (HSV 1 e 2), vírus da varicela zoster (VZV), herpesvirus humano 6 (HHV-6), influenza A (FLUA), enterovírus, vírus da caxumba, poliomavírus vírus BK (BKV) e vírus JC (JCV) para flavivírus. Do total, 44 amostras (36,7%) apresentaram resultado positivo para algum dos vírus analisados no âmbito desta pesquisa, sendo 15 (12,5%) para poliomavírus BKV, 2 (1,7%) para poliomavírus JCV, 21 (17,5%) para HSV1 e 2, 5 (4,2%) foram positivos para BKV e HSV1 e 2 (coinfecção) e 1 (0,8%) para vírus Epstein-Barr (EBV). Uma parte das amostras negativas foi submetida a sequenciamento direto de nova geração (n=8 amostras), resultando em amostras positivas para vírus (vírus simio 40), protozoários e bactérias. Este estudo mostrou que infelizmente, menos de 50% das encefalites e meningites assépticas puderam ser relacionadas a algum agente viral. Houve uma alta prevalência de HSV no material estudado, de acordo com o esperado, mas a presença de poliomavírus no LCR destes indivíduos foi acima da observada na literatura. Esses, bem como os resultados de sequenciamento direto e sua associação a etiologia das encefalites e meningites, devem ser interpretados com cautela. / Central nervous system (CNS) infections caused by microorganism trigger moderate to severe symptoms, depending on the region affected and may be referred as encephalitis or meningitis. Viruses are the most common agents in these infections. The viral agents responsible for these diseases with highest incidence worldwide are certain herpesviruses, flaviviruses, influenza A, enteroviruses, and mumps virus. However, their prevalence vary according to the population, immunological state of the individual, age and region studied. Although there are well-established data on the etiology of these diseases in some countries, there is little information regarding the etiology of these diseases in Brazil. Thus, data regarding the prevalence of these agents in our environment is necessary for the development and application of faster and more efficient diagnostic methods. In this study, 120 cerebrospinal fluid (CSF) samples from two centers of the city of São Paulo (Hospital Santa Casa de Misericordia and Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo) were investigated by PCRs for herpes simplex virus (HSV 1 and 2), varicela zoster virus (VZV), human herpesvirus 6 (HHV6), influenza A, enterovirus, mumps virus, polyomavirus BK virus and JC virus and flaviviruses. From these, 44 samples (36.7%) presented positive result for one of the viruses analyzed, being 15 (12.5%) for polyomavirus BKV, 2 (1.7%) for polyomavirus JCV, 21 (17.5%) for HSV 1 and 2, 5 (4.2%) samples were positive for BKV and HSV1 and 2 (coinfection) and 1 (0.8%) for Epstein-Barr virus (EBV). A part of the negative samples (n=8) were submitted to next generation direct sequencing and revealed the presence of agents as viruses (simian virus 40), protozoa and bacteria. This study showed that unfortunately, less than 50% of the aseptic encephalitis and meningitis could be related to some viral agent. It was found high prevalence of HSV, as expected, but the presence of polyomavirus in the CSF of these individuals was higher than that observed in the literature. These results, as well as direct sequencing results and its relationship to the etiology of encephalitis and meningitis should be interpreted with caution.

Para avaliar a fagocitose de células da série monócitos-macrófagos de cabras naturalmente infectadas pelo vírus da artrite encefalite caprina a vírus (VAEC), 30 cabras da raça Saanen foram utilizadas e alocadas em dois grupos distintos, sendo um grupo formado por 15 animais soropositivos à imunodifusão em gel de ágar para detecção de anticorpos séricos antivírus da AEC e o outro formado por 15 animais soronegativos ao teste. Células mononucleares de sangue periférico foram isoladas por gradiente de concentração e plaqueadas em placas de poliestireno para isolamento de células da série monócito-macrófago. Após adesão das mesmas, adicionou-se a bactéria Corynebacterium pseudotuberculosis como desafio antigênico. Dois tipos de fagocitose foram observados em relação ao número de bactérias internalizadas nos vacúolos citoplasmáticos dos macrófagos, isto é, verificou-se a presença de células fagocitando em média 12 bactérias e outro grupo fagocitando um número incontável de bactérias. Dessa forma, classificou-se a fagocitose em fagocitose + (até 12 bactérias) e fagocitose ++ (mais de 12 bactérias). Em relação à fagocitose total (fagocitose + e fagocitose ++) não foram verificadas diferenças estatística significativas entre os grupos experimentais (p < 0,41). Todavia, ao discriminar a fagocitose conforme a quantificação de bactérias fagocitadas, encontrou-se diferença estatística entre os grupos, sendo que o grupo positivo apresentou maior porcentagem de fagocitose ++ (p < 0,001). O grupo negativo apresentou maior porcentagem de fagocitose + (p < 0,012). Correlação positiva entre monócitos fagocitando e fagocitose ++ foi observada no grupo de animais soropositivos (p < 0,006), porém, não foi observada nenhuma correlação no grupo negativo (p < 0,49). Esses resultados demonstram uma possível alteração na intensidade da fagocitose de macrófagos de animais infectados com o VAEC, sugerindo que os animais com artrite encefalite caprina estejam mais susceptíveis à linfadenite caseosa. / To evaluate the phagocytosis from monocyte-macrophage line cells 30 naturally infected Saanen goats with caprine arthritis-encephalitis vírus (VCAE) were used, and divided uniformly in different groups according to agar gel immunodiffusion test (AGID). Peripheral Blood Mononuclear Cell (PBMC) was isolated by density gradient centrifugation. The monocyte-macrophage cells were separated from PBMC by their adherence properties. Afterwards, the phagocytosis function was assessed by phagocytosis assay using Corynebacterium psudotuberculosis as a source of antigen. Therefore, two distinct types of phagocytosis were observed and were set according to the number of bacteria within the cytoplasmatic vacuoles. Thus, the phagocytosis rates were also divided in two groups, on the first was observed up to 12 bacteria in the vacuoles; on the other hand in the second group an uncountable number of bacteria were usually seen. Consequently, the phagocytosis rates were also divided in phagocytosis + (up to 12 bacteria) and phagocytosis ++ (more than 12 bacteria). The results from the phagocytosis rates show any difference, however when the phagocytosis rates were separated in the order of the number of Corynebacterium pseudotuberculosis phagocyted, the phagocytosis ++ from positive animals in the sera test were higher than the negative one (p < 0.001). Nevertheless, the negative group presents higher pahgocytosis + (p < 0.012). Furthermore a positive and significant correlation between phagocytosis ++ and monocyte phagocytosis (p < 0.006) were also encountered in the positive animals, however the same were not observed in the negative group (p < 0.49). In face of, the results from the present trial point out to higher susceptibility to caseous linphadenitis in goats infected with VCAE due to the alteration on the phagocytosis strength in these animals.

O objetivo do presente estudo foi estabelecer os valores de referência do proteinograma de soro lácteo por meio da técnica de eletroforese SDS-PAGE para a lactação plena e avaliar os efeitos do processo de secagem da glândula mamária, fase colostral e primeiro mês de lactação, fase de lactação, número de lactações, isolamento bacteriano e infecção pelo VCAE nas proteínas da secreção láctea de cabras da raça Saanen. Foram analisadas, entre 2007 e 2010, 545 amostras de leite provenientes de 185 cabras em diversas fases da lactação. Durante a lactação plena, baseado nos resultados dos intervalos de confiança, foram determinados os seguintes valores de referência: proteína total entre 2.940,0 e 3.050 mg/dL; proteína do soro lácteo entre 903,0 e 973,0 mg/dL; lactoferrina entre 68,0 e 77,0 mg/dL; albumina entre 88,0 e 97,0 mg/dL; imunoglobulina cadeia pesada entre 93,3 e 103,0 mg/dL; imunoglobulina cadeia leve entre 132,7 e 146,0 mg/dL; &beta;-lactoglobulina entre 299,0 e 329,0 mg/dL e &alpha;-lactoalbumina entre 213,0 e 229,5 mg/dL. Os valores absolutos de proteína total, proteína do soro e frações protéicas aumentam durante a secagem da glândula. Antes da secagem predominavam as frações de &beta;-Lg e &alpha;-La, a partir do 3&ordm; dia, ocorre o surgimento das novas frações e a alteração do perfil protéico sem que haja o predomínio de nenhuma fração. A fase colostral, primeiras 24 horas de lactação, determinam as maiores concentrações de proteína total, proteína do soro e frações protéicas que diminuem após as primeiras 12 horas de lactação estabilizando após o 5&ordm; dia. No colostro as imunoglobulinas são predominantes, e após o período de transição do colostro para o leite as frações &beta;-Lg e &alpha;-La são predominantes. Nos primeiros 15 dias de lactação, devido à influência da fase colostral, observa-se que as concentrações de proteína total e proteína do soro lácteo são maiores. A partir desse momento permanecem estáveis voltando a aumentar no final da lactação. As frações protéicas do soro de leite (lactoferrina, albumina sérica, imunoglobulina de cadeia pesada, imunoglobulina de cadeia leve, &beta;-Lg e &alpha;-La) também são máximas nos primeiros 15 dias de lactação e diminuem ao longo do período. A concentração de proteína do soro e suas frações em cabras primíparas foi menor quando comparadas com cabras pluríparas. O isolamento bacteriano não influencia as concentrações de proteína total do leite e proteína do soro lácteo de cabras, contudo a concentração de lactoferrina é maior e as concentrações de &beta;-Lg e &alpha;-La são menores em amostras com isolamento bacteriano. O CAEV não influencia as concentrações de proteína total do leite e proteína do soro lácteo de cabras, contudo a concentração de lactoferrina é maior e a concentração de e &alpha;-La é menor em cabras sororeagentes positivas ao VCAE. / The aim of this study was to establish reference values of the whey protein through the technique of SDS-PAGE for the full lactation and to evaluate the effects of the dry period of the mammary gland, colostral phase and first month of lactation, lactation, lactation number, bacterial isolation and VCAE infection in proteins of milk secretion in Saanen goats. Were analyzed between 2007 and 2010, 545 milk samples from 185 goats at different stages of lactation. During full lactation, based on the results of the confidence intervals were determined the following reference values: total protein between 2,940.0 and 3,050 mg / dL; whey protein between 903.0 and 973.0 mg / dL; lactoferrin between 68.0 and 77.0 mg / dL, serum albumin between 88.0 and 97.0 mg /dL, immunoglobulin heavy chain between 93.3 and 103.0 mg / dL, immunoglobulin light chain between 132.7 and 146, 0 mg / dL, &beta;-lactoglobulin between 299.0 and 329.0 mg / dL and &alpha;-lactalbumin between 213.0 and 229.5 mg / dL. The absolute values of total protein, whey protein and protein fractions increase during the dry period. Prevailed prior to dry period the fractions of &beta;-Lg and &alpha;-La from the 3rd day, occurs the emergence of new fractions and protein profile changes without the predominance of any fraction The colostral phase, the first 24 hours of lactation, determine the highest concentrations of total protein, whey protein and protein fractions that decrease after the first 12 hours of lactation stabilized after the 5th day. Immunoglobulin in colostrum is prevalent, and after the period of transition from colostrum to milk fractions &beta;-Lg and &alpha;-La are predominant. In the first 15 days of lactation, due to the influence of colostral phase, it is observed that the concentrations of total protein and whey protein are higher. From then remain stable before rising again in late lactation. The protein fractions of whey (lactoferrin, serum albumin, immunoglobulin heavy chain, immunoglobulin light chain, &beta;-Lg and &alpha;-La) are also maximal in the first 15 days of lactation and decrease during the period. The concentration of whey protein and protein fractions in heifers are smaller when compared with multiparous goats. Bacteria isolation does not influence the concentrations of total protein from milk and whey protein of goats, but the concentration of lactoferrin is increased and the concentrations of &beta;-Lg and &alpha;-La is smaller in samples with bacterial isolation. The CAEV does not influence the concentrations of total protein and whey protein in goat, but the concentration of lactoferrin is higher and concentration of &alpha;-La is less in goat positive by the CAEV.

Diante da ausência de tratamentos eficazes para a esclerose múltipla (EM) e sabendo que compostos animais têm sido usados como protótipos para o desenvolvimento de novas drogas, avaliamos o efeito do Tnp, um peptídeo cíclico inédito e com potencial antiinflamatório derivado do veneno de Thalassophryne nattereri, na encefalomielite autoimune experimental (EAE), um modelo representativo da EM. Demonstramos que o Tnp em distintos esquemas de tratamento por mecanismos também dependentes de IL-10 consegue diminuir a intensidade dos sintomas clínicos e adiar o pico de aparecimento dos sintomas graves na EAE por suprimir DC convencionais e propiciar DC plasmocitóides; por bloquear a infiltração de leucócitos e a reativação de clones Th1, Th17, microglia e macrófagos no SNC; por favorecer o aumento de células reguladoras e ainda por ultrapassar a BHE e alcançar o órgão alvo. O Tnp atenua a neuroinflamação e previne a desmielinização, refletindo assim na melhoria dos sinais clínicos na EAE, tornando-se um importante candidato para o tratamento da EM. / Given the lack of effective treatments for multiple sclerosis (MS) and knowing that venomous have been used as prototype for the development of new drugs here we evaluated the effect of the Tnp, a described antiinflammatory cyclic peptide identified in the venom of Thalassophryne nattereri, on experimental autoimmune encephalomyelitis (EAE), a representative model of MS. We found that distinct treatments of Tnp by mechanisms also dependent on IL-10 significantly reduced the clinical severity of EAE. Tnp was related with: suppression of the activation state of conventional DC and the emergence of plasmacytoid DC; blocking the leukocyte infiltration and the reactivation of Th1, Th17, microglia cells and macrophages in the CNS; increasing of regulatory cells and also Tnp can overcome the BBB and reach the target organ. Tnp can reduce the severity of symptoms and delay the peak of onset of severe symptoms. These results shed light on the role of Tnp a small molecule in the regulation of inflammation and provides a new therapeutic opportunity for the treatment of MS.