No presente trabalho procura-se caracterizar biológica e ecologicamente o Aedes albopictus (Skuse) (Diptera: Culicidae) no seu mais recente ambiente geográfico, isto é, nas Américas. São descritos aspectos relacionados com a chegada e dispersão da espécie no Continente Americano. Paralelamente, são discutidos aspectos relativos ao comportamento, biologia, bionomia, ecologia, hematofagia, interação com outras espécies do gênero Aedes, bem como a importância epidemiológica do Ae. albopictus. Esses aspectos foram descritos, por serem considerados importantes pela influência que exercem na competência vetora da espécie, bem como na sua adaptação ao ambiente antrópico. Ressaltando a importância epidemiológica deste vetor para a espécie humana, citam-se dados referentes à potencialidade do Ae. albopictus participar na transmissão de arboviroses. Assim, procura-se caracterizar a importância da possível participação do Ae. albopictus na veiculação do vírus dengue, febre amarela, bem como outros arbovírus que afetam o ser humano nas Américas. Paralelamente, discute-se a possibilidade do Ae. albopictus atuar como vetor da Dirofilaria immitis (Leidy) ao homem. / In the present work, biological and ecological features of Aedes albopictus (Skuse) (Diptera: Culicidae) are discussed based on literature records. The main focus of this study is the epidemiological importance of the presence of this species in Americas. Aspects relative to colonization and dispersion of Ae. albopictus in Americas are discussed. Dispersion, ethology, biology, ecology, haematophagy, bionomics as well as epidemiological importance, and relationships among Ae. albopictus and other members of the genus Aedes are discussed. These aspects are included because of their influence in the vector competence of Ae. albopictus, and also in its adaptation to the human environment. Because of the epidemiological importance of Ae. albopictus to humans, data relative to the potentiality of this species to participate in the transmission of arboviruses are discussed. In this sense, the importance of Ae. albopictus as vector of dengue, yellow fever, and other arboviruses that cause human diseases is described. Besides, it is discussed the potentiality of Ae. albopictus to act as a vector of Dirofilaria immitis (Leidy) to humans.

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2016-05-11T14:27:17Z No. of bitstreams: 1 PDF - Noiana de Paula Noia.pdf: 1156392 bytes, checksum: a6a9764178b9ec1cd59f8dadd9cb363e (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-07-22T13:24:22Z (GMT) No. of bitstreams: 1 PDF - Noiana de Paula Noia.pdf: 1156392 bytes, checksum: a6a9764178b9ec1cd59f8dadd9cb363e (MD5) / Made available in DSpace on 2016-07-22T13:24:22Z (GMT). No. of bitstreams: 1 PDF - Noiana de Paula Noia.pdf: 1156392 bytes, checksum: a6a9764178b9ec1cd59f8dadd9cb363e (MD5) Previous issue date: 2013-12-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The Aedes genus is of great epidemiological importance, highlighting the Aedes aegypti Linnaeus, 1762 and Aedes albopictus Skuse, 1894, the main vectors of dengue, yellow fever and other arboviruses. How are spread across the planet, have become a public health risk factors. Aimed to evaluate the effects of temperature on competition and in the life cycle, and the source of blood meal in fecundity and longevity of females of A. aegypti and A. albopictus. The collections of eggs were performed using traps ovitraps type. The experiments were performed at the Laboratory of Entomology, University of Paraíba State - UEPB in Campina Grande, PB. Competition between A. aegypti and A. albopictus was examined at temperatures of 20, 24, 28, 32 and 36 ° C in the proportion of 10:0, 7:3, 5:5, 3:7 and 10:0 (A. aegypti : A. albopictus). The cycle life was examined at the same temperatures. Feeding two treatments with each meal in quail and the other in swiss mice were used. For all experiments we used 12h light and 70% humidity. In interspecific relationship, there was overlap of A. aegypti on A. albopictus from the first generation (F1), independent of temperature and the proportion of species. The long life cycle did not differ significantly among species independent of temperature, but decreased with increasing temperature. The base temperature (Tb) and thermal constant (K) and hence the number of generations was greater for A. albopictus than for A. aegypti. The repast in mice resulted in greater number of eggs and female longevity for both A. aegypti as for A. albopictus. The substrate used for biting that was more appropriate for the two species, the mouse was, since we obtained the highest female fertility and longevity. The temperature and the substrate used for blood feeding interfere with competition, development, longevity and fecundity of the species studied. The number of generations of insects in the laboratory is higher for A. albopictus than for A. aegypti. / O grande importância epidemiológica, destacando o Aedes aegypti Linnaeus, 1762 e Aedes albopictus Skuse, 1894, principais vetores da dengue, febre amarela e outras arboviroses. Como estão distribuídos por todo o planeta, se tornaram fatores de risco à saúde pública. Objetivou-se avaliar os efeitos da temperatura na competição e no ciclo de vida, e da fonte de repasto sanguíneo na fecundidade e longevidade de fêmeas de A. aegypti e A. albopictus. As coletas dos ovos foram realizadas utilizando armadilhas do tipo ovitraps. Os experimentos foram realizados no Laboratório de Entomologia da Universidade Estadual da Paraíba – UEPB, em Campina Grande, PB. A competição entre A. aegypti e A. albopictus foi analisada nas temperaturas de 20, 24, 28, 32 e 36ºC nas proporções de 10:0, 7:3, 5:5, 3:7 e 10:0 (A. aegypti: A. albopictus). O ciclo de vida foi analisado nas mesmas temperaturas. Para nutrição foram utilizados dois tratamentos um com o repasto em codorna e o outro em camundongo swiss. Para todos os experimento utilizou-se fotofase de 12h e umidade de 70%. Na relação interespecífica, constatou-se a sobreposição de A. aegypti sobre A. albopictus a partir da primeira geração (F1), independente da temperatura e da proporção das espécies. O tempo do ciclo de vida não diferiu significativamente entre as espécies, independente da temperatura, porem diminuiu em função do aumento da temperatura. A temperatura base (Tb) e a constante térmica (K) e consequentemente o número de gerações anuais foi maior para A. albopictus do que para A. aegypti. O repasto em camundongos resultou em maior número de ovos e longevidade das fêmeas tanto para A. aegypti como para A. albopictus. O substrato utilizado para hematofagia que se mostrou mais adequado para as duas espécies, foi o camundongo, visto que obteve-se a maior fertilidade e longevidade da fêmea. A temperatura e o substrato utilizado para o repasto sanguíneo interferem na competição, desenvolvimento, longevidade e fecundidade das espécies estudadas. O número de gerações anuais dos insetos em laboratório é maior para A. albopictus do que para A. aegypti.

West Nile is an important novel virus in the United States, having spread rapidly since it was first detected in New York in 1999. The Centers for Disease Control and Prevention as well as many State Health Departments, have mandated programs for surveillance of West Nile Virus activity. These programs incorporate many different aspects including existing arboserology programs with additional testing for West Nile Virus and new plans that incorporate active and passive surveillance methods. The objective of this study was to examine all aspects of the Florida West Nile surveillance program to determine if there was transmission in the animal systems prior to human cases. The predictive analyses were done using regional data graphs, spatial information, correlations and regression models. Data for sentinel chickens, bird necropsy and mosquito pool surveillance from participating counties in Florida were obtained from the State of Florida surveillance database. The human data was obtained from the State of Florida reportable disease database for each county whether participating in the state surveillance programs or not. Clinical cases were examined by demographics (gender and age) and an incidence rate was calculated to demonstrate the effects of disease. Specific statistical methods used included Pearson's coefficient correlation, Poisson distribution regression modeling to show if any of the surveillance systems were predictors for human disease. The incidence rate analysis for clinical cases showed clustering of cases in adjacent counties within a region where Florida's panhandle and adjacent counties northeast had the highest incidence. Florida's central and southern regions had moderate human incidence. This provides useful information in transmission geography for prevention and control measures. Demographic analysis showed that there were twice as many males than females diagnosed with West Nile in Florida, this was true across the groups as well. The highest number of cases was seen within the age group over 55 years of age for West Nile Neuroinvasive Disease and for West Nile Fever the highest number of cases was within the 36-54 age range. The temporal distribution was determined using graphical representations of all of the surveillance types and clinical cases. In order to include all relevant data, the temporality was set from week 20 to week 52. This study found that all of the surveillance types (dead birds, mosquitoes and sentinels) offered a specialized strength for predicting clinical cases. However, mosquitoes proved to be the least efficient out of the three surveillance systems. The regional and spatial analysis showed that positive dead birds and sentinels provided the coverage for the surveillance systems in the state. However, Pearson's correlation coefficient was low for sentinel surveillance; this may be due to higher participation showing West Nile Virus activity in areas (especially rural) that have no reported human cases. This analysis did show that West Nile is detected in mosquito pool samples before it is detected in the dead bird or sentinel surveillance systems which provides an earlier warning for human cases. The Poisson distribution regression model was only useful for the pooled years and 2003. These showed that mosquitoes, positive dead birds and sentinels were good predictors for clinical cases for the combined years and dead birds and sentinels were significant for 2003 as well. The recommendations based on the results from this study would be to continue all the current surveillance efforts but with the following enhancements: 1. Increase the coverage and consistency of submissions for all surveillance types. 2. Set standard levels of participation for all counties based on the regional analyses and populations at risk. 3. Create standardized approaches for sampling, shipping and submitting samples (especially for mosquito pool submissions) and require that participating counties adhere to these standards. 4. Only submit specific birds known to be especially susceptible to West Nile Virus (e.g. corvids). 5. Targeted prevention and education strategies for higher risk groups based on their potential levels of exposure.

Florida's mild climate supports year round enzootic transmission of arthropod-borne (arbo) viruses, such as St. Louis Encephalitis virus (SLEV), West Nile virus (WNV), and Eastern Equine Encephalomyelitis virus (EEEV). First isolated in 1960 from two Florida blue jays, Highlands J virus (HJV) is endemic to the state and vectored by the same mosquitoes as EEEV (Henderson et al, 1962). EEEV and HJV are both alphaviruses, but HJV is not pathogenic to humans, occasionally causes encephalitis in horses, and is a recognized pathogen in some bird species (turkeys, emus, etc) (Cilnis et al, 1996). The Florida Sentinel Chicken Arboviral Surveillance Program, established in 1978, utilizes sentinel chickens to detect arboviral activity throughout the state. Current serologic antibody detection methods include the hemagglutination inhibition (HAI), IgM antibody capture enzyme-linked immunosorbent (MAC-ELISA), and serum neutralization plaque reduction (PRNT) assays (Blackmore et al, 2003). In 2003, sentinel chickens detected significantly greater alphavirus activity than seen in the previous 15 years (Stark & Kazanis, 2003). This increase raised concerns that bridging into the human population had become a serious threat as well as an important issue for veterinary health. The objective of this study was to determine if cross-reactions with Highlands J virus were impacting the serologic diagnostic tests routinely performed for identification of EEEV. For 2003, the HAI test detected 476 alphavirus positive sentinels. We tested 316 of these chickens in the PRNT, which identified 176 EEEV positive sentinels and 75 HJV positive sentinels. Our results indicate that Highlands J virus is extensively cross-reactive in the HAI test and that the MAC-ELISA is more specific for the detection of antibodies solely to EEEV. We demonstrated that EEEV antibody titers in the HAI test were positively correlated to antibody titers in the PRNT assay. Analysis of alphaviral activity by county indicates widespread transmission of HJV across the northern and panhandle regions of the state; however EEEV activity was greater than HJV activity in all but four counties. Consequently, distinguishing between the two agents can reduce the expenditure of resources on unnecessary vector control and medical alerts to protect the public health from Highlands J virus.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2017-05-03T15:18:12Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) FERREIRA, H. S._Dissertação_FINAL.pdf: 9243519 bytes, checksum: f2021a9042905cd6559a1e42abc0aa57 (MD5) / Made available in DSpace on 2017-05-03T15:18:12Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) FERREIRA, H. S._Dissertação_FINAL.pdf: 9243519 bytes, checksum: f2021a9042905cd6559a1e42abc0aa57 (MD5) Previous issue date: 2016-02-29 / FACEPE / O planejamento urbano atrelado às perspectivas de desenvolvimento sustentável podem minimizar os riscos socioambientais urbanos que se consolidaram na maioria das grandes cidades brasileiras, dentre eles, os ricos à saúde. As repercussões locais das alterações climáticas observadas na atualidade têm provocado mudanças nos padrões dos elementos atmosféricos, potencializando ainda mais os riscos à saúde, especialmente a expansão e a persistência de surtos epidêmicos de arboviroses como a dengue. O objetivo deste trabalho foi analisar a relação entre a dinâmica do clima urbano e a ocorrência de surtos e formação de epidemias de dengue na cidade do Recife – PE. Foram analisadas as características do clima urbano do Recife – variáveis atmosféricas – temperaturas e umidade relativa do ar, as influencias dos elementos atmosféricos sobre a formação das epidemias e o padrão de distribuição espacial das notificações em relação à indicadores socioeconômicos e físicoambientais. Conforme resultados encontrados verificou-se que os surtos epidêmicos mais expressivos na cidade estão relacionados com as ocorrências de precipitações no primeiro semestre do ano, especialmente durante a quadra chuvosa, sendo possível a identificação de bairros mais e menos suscetíveis aos surtos da doença em períodos com condições atmosféricas favoráveis a reprodução e proliferação do mosquito vetor da dengue. As áreas de morros das zonas norte e sul apresentaram maior problemática em relação a situação da dengue no município, com persistência importante de surtos epidêmicos. A ligação entre os surtos de dengue e as condições climáticas locais, faz das condições atmosféricas um fator digno de vigilância e monitorização como parte das estratégias de combate e controle das epidemias de dengue na cidade. / Urban planning linked to sustainable development prospects can minimize the environmental risks that have been consolidated in most of the Brazilian cities. One of these risks are the health risks. Nowadays, local effects of climate change can be observed, and they are leading to changes in atmospheric elements patterns, which in turn enhance health risks even more. Among these risks, we highlight the expansion and persistence of arboviral diseases outbreaks, such as dengue episodes. The objective of this study was to analyze the relationship between urban climate dynamics and occurrences of dengue epidemics outbreaks in the city of Recife, PE. We analyzed the characteristics of the urban climate of Recife (temperature and relative humidity), the influence of atmospheric elements on formation of epidemic outbreaks and its spatial distribution regarding socioeconomic, physical and environmental indicators. Results indicated that the most significant outbreaks in the city are related to the occurrence of rainfall in the first half of the year, especially during the rainy season. In addition, it is possible to identify more or less susceptible neighborhoods to disease outbreaks in the periods of favorable weather conditions to reproduction and proliferation of the mosquito. The areas of hills in the northern and southern areas have greater problems regarding the situation of dengue than other areas in the city, showing recurrence of outbreaks. The connection between the dengue outbreaks and the local weather conditions makes weather conditions a decent factor of surveillance and monitoring as part of combat strategies and control of dengue epidemics in the city.

Made available in DSpace on 2015-04-20T12:31:38Z (GMT). No. of bitstreams: 1 tese doutorado total.pdf: 1236976 bytes, checksum: e7099549621a6ea8ce476ffa5cba2cc1 (MD5) Previous issue date: 2007-04-20 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The arboviruses represent serious problems of public health in Brazil, in particular in the Amazon area. Four arboviruses have shown to be particularly important, namely dengue, yellow, Oropouche and Mayaro, since they cause severe human disease with high mortalities and are involved in periodic outbreaks. The mapping of the main arboviruses in the Rondonia State is necessary, as well as the knowledge of their biological cycles, in order to define control measures with the understanding of their period and mechanisms of transmission. It is also necessary to improve diagnostic procedures by viral isolation and molecular characterization by RT-PCR, with the primers selected for identification of these viruses, directly from the serum or tissue fluids of patients, or indirectly from cell cultures inoculated with patient s materials. In the present manuscript are presented results of arbovirus isolation and characterization procedures from two hundred and sixty six samples of blood colleted from patients with suspected of arboviruses infections in the following Rondonia State s cities: Ariquemes, Cacoal, Colorado D´Oeste, Jarú, Ouro Preto D´Oeste, Porto Velho, Theobroma and Vilhena. The samples were inoculated in C6/36 monolayers and after seven days the cellular supernatants were collected for RNA extraction by Trizol® method. The RNA extracted was used for RT-PCR with primers selected from the specialized literature s data. The universal primer was selected for identify Flavivirus, Alphavirus genera and Simbu serogroup. Specific primers were also used to identify the four serotypes of dengue virus and yellow fever virus. The amplicons were submitted to electrophoresis in agarose gel 1,7% and visualized in ultra violet light. For genetic sequencing was used the interruption of extention of chain of nucleotide by dideoxynucleotide technique. The ABI PRISM 310 Genetic Analyser (PE Biosyntesis) was used for sequencing the dengue virus serotype 1, the Personal Seq 4X4 (Amersham Pharmacia Biotech, USA), for sequencing the dengue virus serotype 3 and the Mega BACE1000 (Amersham Pharmacia Biotech, USA) for sequencing the Cacipacore virus. One hundred fourteen samples were indentifyed by RT-PCR. The results of identification of positive collected samples from different periods of 2003 to 2005 was the following: 70 patients with dengue serotype 1, one patient with dengue serotype 2, 42 patients with dengue serotype 3 and one patient com Cacipacore virus. The genetic sequencing performed of dengue virus serotypes 1 and 3 samples analyzed by software Clustal W showed 98% of similarity with others dengue virus isolated and sequenced in southeastern areas of Brazil. The Cacipacore virus sample isolated fom the Theobroma patient was the first sample isolated from human infections and presented 91% of similarity with others virus samples isolated in Brazil from migratory birds. We concluded from, our studies, that the epidemiologic surveillance, using molecular methods associated to serological ones is more accurate and precise than serological methods alone. It must be added the easiness of the procedure and the speed for getting the result in contrast with complicated methodologies like hemaglutination inhibition or complement fixation methodologies that need double samples and eventually give cross-reacting results with others arboviruses. The selected primers, for Multiplex-RT-PCR and for nested-RT-PCR used in this work, identify the main arboviruses circulating not only in the Rondonia State that can be used in other centers of research in the region north region of Brazil. Our results also emphasize the agility of the method consisting in the direct use of patient s serum for the RT-PCR procedure which reduces the time for getting a diagnosis. / As arboviroses representam graves problemas de saúde pública no Brasil, particularmente na Região Amazônica. Quatro arboviroses têm-se mostrado especialmente relevantes, a saber: dengue, febre amarela, Oropouche e Mayaro, pois causam doenças humanas graves, com mortalidade relativamente elevada e são responsáveis por surtos epidêmicos. O mapeamento e caracterização dos principais arbovírus registrados no Estado de Rondônia são necessários para identificar quais estão circulando, definir o período de transmissão e seu diagnóstico através de isolamento viral e caracterização molecular pela técnica de RT-PCR, Isso foi realizado neste estudo com a seleção de primers para a identificação desses vírus tanto isolados a partir do soro do paciente quanto a partir de culturas celulares semeadas com soro dos pacientes. No presente trabalho, é apresentado resultado de isolamento e caracterização de arbovírus a partir de duzentas e sessenta e seis amostras de sangue coletadas de pacientes com suspeitas de arbovirose nas cidades de Ariquemes, Cacoal, Colorado D Oeste, Jarú, Ouro Preto D Oeste, Porto Velho, Theobroma e Vilhena. As amostras foram inoculadas em culturas de células C6/36 e após sete dias os fluídos celulares foram recolhidos e submetidos à extração de RNA pelo método do TRIzol®. O RNA extraído foi utilizado para RT-PCR com os primers selecionados na literatura especializada. Os primers universais para Flavivirus, Alphavirus, para os quatro sorotipos de dengue, para o sorogrupo Simbu e especifico para febre amarela foram testados nessas amostras. Os amplicons foram submetidos à corrida eletroforética em géis de agarose 1,7% e visualizados em transiluminador ultravioleta. Para o seqüencimento gênico utilizou-se a técnica de interrupção da cadeia por dideoxinucleotídios. Foram utilizados os seqüenciadores ABI PRISM 310 Genetic Analyzer (PE Biosynthesis, USA), para seqüenciar o vírus dengue sorotipo 1, o Personal Seq 4X4 (Amersham Pharmacia Biotech, USA), para seqüenciar o genoma do vírus da dengue sorotipo 3 e o Mega BACE 1000 (Amersham Pharmacia Biotech, USA), para seqüenciar o vírus Cacipacoré. Cento e quatorze amostras foram identificadas após a realização da RT-PCR. Os resultados de isolamento e caracterização das amostras virais foi o seguinte: 70 pacientes com dengue sorotipo 1, um paciente com dengue sorotipo 2; 42 pacientes com dengue sorotipo 3 e um paciente com o vírus Cacipacoré. A seqüência gênica dos vírus da dengue sorotipos 1 e 3 foram analisadas pelo programa Clustal W e apresentaram 98% de semelhança com aqueles isolados em regiões do sudeste do Brasil. Note-se que a amostra isolada de vírus Cacipacoré representa o primeiro isolamento do vírus de seres humanos e seu seqüenciamento apresentou 91% de similaridade com o de amostras isoladas precedentemente de aves migratórias selvagens. Concluiu-se nestes estudos que a vigilância epidemiológica, associando métodos moleculares e sorológicos de caracterização viral são mais precisos do que aquela feita apenas por técnicas sorológicas. Soma-se a isso o grau de relativa facilidade para se obter o resultado, não sendo uma metodologia que implica em interpretação complexa como a Inibição da Hemaglutinação ou Fixação do Complemento, técnicas que necessitam de amostras pareadas e eventualmente podem fornecer reações cruzadas entre diferentes viroses. Os primers escolhidos tanto para Multiplex-RT-PCR, quanto para Nested-RT-PCR usados neste trabalho, identificam as principais arboviroses circulantes no Estado de Rondônia, e podem ser usados em outros centros de pesquisas na região norte. Os resultados destes estudos mostram que a agilidade do uso direto do soro de pacientes com suspeitas de arboviroses como fonte de material para a RT-PCR recomendam o uso dessa técnica, pois diminui o tempo na obtenção do resultado.

No presente trabalho procura-se caracterizar biológica e ecologicamente o Aedes albopictus (Skuse) (Diptera: Culicidae) no seu mais recente ambiente geográfico, isto é, nas Américas. São descritos aspectos relacionados com a chegada e dispersão da espécie no Continente Americano. Paralelamente, são discutidos aspectos relativos ao comportamento, biologia, bionomia, ecologia, hematofagia, interação com outras espécies do gênero Aedes, bem como a importância epidemiológica do Ae. albopictus. Esses aspectos foram descritos, por serem considerados importantes pela influência que exercem na competência vetora da espécie, bem como na sua adaptação ao ambiente antrópico. Ressaltando a importância epidemiológica deste vetor para a espécie humana, citam-se dados referentes à potencialidade do Ae. albopictus participar na transmissão de arboviroses. Assim, procura-se caracterizar a importância da possível participação do Ae. albopictus na veiculação do vírus dengue, febre amarela, bem como outros arbovírus que afetam o ser humano nas Américas. Paralelamente, discute-se a possibilidade do Ae. albopictus atuar como vetor da Dirofilaria immitis (Leidy) ao homem. / In the present work, biological and ecological features of Aedes albopictus (Skuse) (Diptera: Culicidae) are discussed based on literature records. The main focus of this study is the epidemiological importance of the presence of this species in Americas. Aspects relative to colonization and dispersion of Ae. albopictus in Americas are discussed. Dispersion, ethology, biology, ecology, haematophagy, bionomics as well as epidemiological importance, and relationships among Ae. albopictus and other members of the genus Aedes are discussed. These aspects are included because of their influence in the vector competence of Ae. albopictus, and also in its adaptation to the human environment. Because of the epidemiological importance of Ae. albopictus to humans, data relative to the potentiality of this species to participate in the transmission of arboviruses are discussed. In this sense, the importance of Ae. albopictus as vector of dengue, yellow fever, and other arboviruses that cause human diseases is described. Besides, it is discussed the potentiality of Ae. albopictus to act as a vector of Dirofilaria immitis (Leidy) to humans.

Rondônia apresenta área com rica diversidade de artrópodes, porém pouco se conhece sobre a transmissão de arbovírus por estas espécies. O presente trabalho visou detectar arbovírus, por meio da RT-PCR e da Duplex RT-PCR, nas espécies de dipteros coletados no Estado, bem como caracterizá-los pela reação de sequenciamento. A RT-PCR e a Duplex RT-PCR detectaram as suspensões dos vírus Mayaro e Oropouche até 104 e 101 TCID50/mL, respectivamente, porém o vírus Dengue 2 em pools contendo menos de três mosquitos infectados foi negativa. O controle endógeno foi detectado em 66,8 % das amostras, sendo que, em pools contendo entre um e três mosquitos, a detecção foi aproximadamente metade da detecção nos pools contendo entre 11 e 15. Em 0,66 % dos pools foi encontrado o vírus Oropouche e em outros 0,66 %, o vírus Cacipacoré. O vírus Oropouche foi detectado em Coquillettidia sp. e Deinocerites sp., enquanto o vírus Cacipacoré foi encontrado em Anopheles sp. e Culex sp. As técnicas possibilitaram a detecção dos arbovírus pesquisados nos pools coletados em Rondônia. / The Rondônia state has an area with rich arthropods diversity although the knowledge about the arboviruses transmition for these species is poor. The present work aimed to detect arboviruses through RT-PRC and RT-PCR Duplex in the diptera species collected in the region as well as their characterization through the sequence reaction. The RT-PRC and RT-PCR Duplex detected the Mayaro and Oropouche virus suspensions until 104 e 101 TCID50/mL respectively, although it was negative for the Dengue 2 virus in pools containing less than three infected mosquitoes. The endogenous control was detected in 66,8 % of samples and from pools containing from one to three mosquitoes the detection rate was approximately half from that obtained from pools with 11 to 15 mosquitoes. Oropouche virus was found in 0,66 % of pools and Cacipacore virus also in 0,66 % of pools. Oropouche virus was detected in Coquillettidia sp. and Deinocerites sp. while Cacipacoré virus was found in Anopheles sp. and Culex sp. The techniques allowed the detection of examined arboviruses in the pools collected from Rondonia.

Philosophiae Doctor - PhD / Bionomics of vector-borne pathogens (VBPs) is a complex phenomenon that involves understanding the ecology of arthropod borne pathogens and vertebrate hosts potentially involved in their transmission cycles. Investigations into the bionomics of viral and bacterial VBPs circulating in Baringo and Homa Bay Counties of Kenya were carried out. Specifically, vertebrate hosts represented in mosquito bloodmeals, presence of arboviruses in blood fed mosquitoes and patients presenting with acute undiagnosed febrile illnesses in rural health facilities, and tick borne pathogens (TBPs) diversity in ticks of animals were identified. Mosquitoes were trapped by BG sentinel and CDC light traps, while ticks were sampled directly from domestic animals and tortoises close to human habitation along the shores and adjacent islands of Lakes Victoria and Baringo in Kenya. Blood and sera were also sampled from patients presenting with acute febrile illnesses visiting four rural health facilities in Homa Bay County. Mosquitoes and ticks were sorted and identified to species using standard morphological taxonomic keys. All the biological samples (blood-fed mosquitoes, ticks and blood/sera) were processed using molecular and culture procedures for detection of VBPs (arboviruses, Ehrlichia, Anaplasma, Rickettsia and protozoa). Among 445 blood-fed Aedeomyia, Aedes, Anopheles, Culex, Mansonia, and Mimomyia mosquitoes, 33 bloodmeal hosts were identified including humans, eight domestic animal species, six peridomestic animal species and 18 wildlife species. Further detection of Sindbis and Bunyamwera viruses was done on blood-fed mosquito homogenates by Vero cell culture and RTPCR in Culex, Aedeomyia, Anopheles and Mansonia mosquitoes from Baringo that had fed on humans and livestock. In TBPs assay, 585 tick pools were analysed consisting of 4,126 ticks collected in both study areas. More ticks were sampled in Baringo (80.5%), compared to Homa Bay (19.5%). In Baringo, agents of ehrlichiosis were detected from Amblyomma and Rhipicephalus ticks including Ehrlichia ruminantium (12.3%), Ehrichia canis (10.5%) and Paracoccus sp. (4.4%). Agents of anaplasmosis included Anaplasma ovis (7.2%), Anaplasma platys (4.4%) and Anaplasma bovis (4.0%), all from Hyalomma, Amblyomma and Rhipicephalus ticks, as well as agents of rickettsiosis, including Rickettsia africae, Rickettsia aeschlimannii, Rickettsia rhipicephali, Rickettsia montanensis and a Rickettsia sp. that was not conclusively characterized. Babesia caballi, Theileria sp. and Hepatozoon fitzsimonsi were also detected from both Rhipicephalus ticks and Amblyomma ticks. In Homa Bay, Ehrichia ruminantium (17.5%) and Ehrichia canis (9.3%) were isolated from Amblyomma latum and Rhipicephalus pulchellus, as well as Anaplasma platys (14.4%) and Anaplasma ovis (14.4%) from Amblyomma and Rhipicephalus species. In determination of the occurrence of arboviruses among patients presenting with acute febrile illnesses, acute Bunyamwera 3 (0.9%) and Sindbis 2 (0.6%) infections were detected by RT-PCR and cell culture and Sindbis seroprevalence was determined by plaque assay. Though a significant proportion of these patients tested positive for low Plasmodium parasitemia, none were co-infected with Plasmodium parasites and arboviruses. This study highlights the presence and relative importance of zoonotic VBPs in both study areas.

Background Arboviral diseases are one of the most common causes of acute febrile illness (AFI) and a significant health problem in South America. In Peru, laboratory etiologic identification of these infections occurs in less than 50% of cases, leading to underdiagnoses of important emerging arboviruses. Aim To assess the prevalence of the Dengue (DENV), Oropouche (OROV), Chikungunya (CHIKV), Mayaro (MAYV) and Zika (ZIKV) viruses in patients with acute febrile illness from Puerto Maldonado (Peru). Methodology Serum samples were obtained from patients with AFI during January 2016 to March 2016. A total of 139 specimens were analyzed for the presence of DENV, OROV, CHIKV, MAYV, and ZIKV using polymerase chain reaction (PCR). Results CHIKV in 9.4% and OROV in 8.6% were the most prevalent arboviruses, followed by DENV and ZIKV, with a prevalence of 6.5% and 5%, respectively. Among all patients, the most common symptoms accompanying fever were headaches 79.9%, muscle pain 65.5% and joint pain 63.3%. Conclusions During this short 3-month period, 4 arboviruses were detected by PCR, CHIKV and OROV being the most common arboviruses in Puerto Maldonado (Peru). Thus, it is crucial to include OROV detection in the national health surveillance. Furthermore, the etiologic clinical diagnosis of arboviral infections is not possible due to the low specificity of symptoms; therefore an increase of cases confirmed by molecular diagnostic methods will enhance arboviral surveillance in Peru.