Thesis (M.S.)--State University of New York at Binghamton, Biology Department, 2006. / Includes bibliographical references.

Accurate identification of pathogens and vectors is essential in epidemiological studies of mosquito-borne pathogens. However, the members of the communis and punctor complexes are difficult to distinguish because they are highly cryptic species, with little to no species-specific morphological characters. The objective of this thesis is to develop molecular tools, including RFLP and DNA barcoding using cytochrome oxidase I (COI), internal transcribed spacer 2 (ITS2) and the intron of ribosomal protein S12 (RPS12) to facilitate identification of the members of these two complexes in Manitoba. A distinct interspecific distance for COI was found between the members of the communis complex included here, and diagnostic RFLP profiles were developed for Oc. communis and Oc. churchillensis. Relatively low average interspecific genetic distances using COI, ITS2 and RPS12 were observed between the members of the punctor complex, indicates no discernable boundaries between these species based on DNA barcoding.

Accurate identification of pathogens and vectors is essential in epidemiological studies of mosquito-borne pathogens. However, the members of the communis and punctor complexes are difficult to distinguish because they are highly cryptic species, with little to no species-specific morphological characters. The objective of this thesis is to develop molecular tools, including RFLP and DNA barcoding using cytochrome oxidase I (COI), internal transcribed spacer 2 (ITS2) and the intron of ribosomal protein S12 (RPS12) to facilitate identification of the members of these two complexes in Manitoba. A distinct interspecific distance for COI was found between the members of the communis complex included here, and diagnostic RFLP profiles were developed for Oc. communis and Oc. churchillensis. Relatively low average interspecific genetic distances using COI, ITS2 and RPS12 were observed between the members of the punctor complex, indicates no discernable boundaries between these species based on DNA barcoding.

Recently, the number of human cases of La Crosse encephalitis (LACE), an illness caused by mosquito-borne La Crosse (LAC) virus, has increased in southwestern Virginia, resulting in a need for better understanding of the virus cycle and the biology of its vectors in the region. This project examined the spatial and temporal distributions of the primary vector of LAC virus, Ochlerotatus triseriatus, and a potential secondary vector, Aedes albopictus. Ovitrapping surveys were conducted in 1998 and 1999 to determine distributions and oviposition habitat preferences of the two species in southwestern Virginia. For virus assay, adult mosquitoes were collected at a tire dump and a human case site during 1998 and 1999, and ovitrap samples were taken from a human case site in 2000. In a separate study, a landcover map of Wise County was created by supervised classification of Landsat Enhanced Thematic Mapper imagery, and maps indicating posterior probabilities of high mosquito abundance were created by combining ovitrap survey-derived, landcover-based prior and conditional probabilities for high and low mosquito abundance using remote sensing techniques and Bayesian decision-making rules. Both Oc. triseriatus and Ae. albopictus were collected from all ovitrap sites surveyed in Wise, Scott, and Lee Counties during 1998. Numbers of Oc. triseriatus remained high from late June through late August, while Ae. albopictus numbers increased gradually through June and July, reaching a peak in late August and declining thereafter. Overall, Oc. triseriatus accounted for 90.1% of eggs collected during this period, and Ae. albopictus made up the remaining 9.9%. Abundance of the two species differed among the sites, and in Wise County, relative Ae. albopictus abundance was highest in sites with traps placed in open residential areas. Lowest numbers of both species were found in densely forested areas. Ovitrapping at a human LACE case site during 1998 and 1999 revealed that Aedes albopictus was well-established and overwintering in the area. An oviposition comparison between yard and adjacent forest at the Duncan Gap human LACE case site in 1999 showed that Ae. albopictus preferentially oviposited in the yard surrounding the home over adjacent forested areas, but Oc. triseriatus showed no preference. LAC virus was isolated from 1 larval and 1 adult collection of Oc. triseriatus females from the Duncan Gap human case site, indicating the occurrence of transovarial transmission at this site. The supervised landcover classification for Wise County yielded a landcover map with an overall accuracy of 98% based on comparison of output classification with user-defined ground truth data. Posterior probability maps for Oc. triseriatus and Ae. albopictus abundance reflected seasonal and spatial fluctuations in mosquito abundance with an accuracy of 55-79% for Oc. triseriatus (Kappa=0.00-0.53) and 70-94% for Ae. albopictus (Kappa=0.00-0.49) when model output was compared with results of an ovitrapping survey. Other accuracy measures were also considered, and suggestions were offered for improvement of the model. / Master of Science

Amostras populacionais de Oc. scapularis foram coletadas nos municípios de Tremembé-SP (TRE), São Paulo-SP (SPA) e Itaboraí-RJ (ITA). Foram empregados como marcadores biológicos: o gene mitocondrial Citocromo Oxidase Subunidade-1 (COI), geometria alar e análise da genitália masculina. Tais marcadores são tradicionalmente reconhecidos pelo poder discriminante em estudos desta natureza. As populações ITA, TRE e SPA mostraram-se distintas quanto à forma alar, sugerindo baixo fluxo gênico entre elas. Foi verificado dimorfismo sexual em relação ao tamanho isométrico, à forma alar e ao grau de diferenciação populacional. A população de ITA apresentou menor tamanho dos centróides que as demais populações estudadas. Foi verificado amplo polimorfismo genético, tendo sido detectados 51 haplótipos de COI e apenas 11 compartilhados entre as populações ITA, TRE e SPA. Quando comparados com as populações de estudo anteriormente realizado por Devicari, 2010 encontramos um padrão parecido. Analisando conjuntamente os presentes dados com aqueles obtidos por Devicari, 2010, computamos 52 haplótipos sendo apenas alguns compartilhados. Os valores do índice de diferenciação genética <font face=\"Symbol\">Fst observados foram moderados somente entre SPA e ITA, nas demais populações estudadas a diferenciação foi baixa, a diferenciação observada foi compatível com a hipótese de distanciamento geográfico das populações coletadas. Analisando cada marcador biológico, concluímos que as populações estudadas não se tratam de complexo de espécies. Ainda não descartamos a existência de complexo dentro de Oc. scapularis, porém, para definitiva resposta a essa questão serão necessários mais estudos envolvendo populações de outras regiões. / Samples of Oc. scapularis were collected in the municipalities of Tremembé-SP (TRE), São Paulo-SP (SPA) and Itaboraí-RJ (ITA). We used the following biological markers: mitochondrial cytochrome oxidase subunit-1 gene (COI), wing geometry and shape of male genitalia. These markers are traditionally known by its discriminating power in studies of this nature. ITA, SPA and TRE populations, showed distinct wing shape, suggesting low gene flow. We observed sexual dimorphism concerning the isometric size, wing shape and the degree of populational differentiation. ITA sample exhibited the lowest centroid sizes. We found high genetic polymorphism in all populational samples, being 51 COI haplotypes. Out of them, only 11 haplotypes were noted to be shared between at least two or three populations. When comparing our results with those of a previous survey conducted by Devicari (2010), we found a quite similar pattern of hign polymorfism. In total, both studies comprised 52 haplotypes. The <font face=\"Symbol\">Fst index of genetic differentiation values were considered \"moderate\" between ITA and SPA and \"low\" in the other comparisons. Present results are consistent with the hypothesis that populations are subjected to isolation by geographical distance. Analyzing together each biologcal marker, we conclude that populations studied do not consist a species complex. We do not rule out the possible occurence of a complex in Oc. scapularis, however, a definitive answer to this question will require further studies and sampling of populations from elsewhere.

Amostras populacionais de Oc. scapularis foram coletadas nos municípios de Tremembé-SP (TRE), São Paulo-SP (SPA) e Itaboraí-RJ (ITA). Foram empregados como marcadores biológicos: o gene mitocondrial Citocromo Oxidase Subunidade-1 (COI), geometria alar e análise da genitália masculina. Tais marcadores são tradicionalmente reconhecidos pelo poder discriminante em estudos desta natureza. As populações ITA, TRE e SPA mostraram-se distintas quanto à forma alar, sugerindo baixo fluxo gênico entre elas. Foi verificado dimorfismo sexual em relação ao tamanho isométrico, à forma alar e ao grau de diferenciação populacional. A população de ITA apresentou menor tamanho dos centróides que as demais populações estudadas. Foi verificado amplo polimorfismo genético, tendo sido detectados 51 haplótipos de COI e apenas 11 compartilhados entre as populações ITA, TRE e SPA. Quando comparados com as populações de estudo anteriormente realizado por Devicari, 2010 encontramos um padrão parecido. Analisando conjuntamente os presentes dados com aqueles obtidos por Devicari, 2010, computamos 52 haplótipos sendo apenas alguns compartilhados. Os valores do índice de diferenciação genética <font face=\"Symbol\">Fst observados foram moderados somente entre SPA e ITA, nas demais populações estudadas a diferenciação foi baixa, a diferenciação observada foi compatível com a hipótese de distanciamento geográfico das populações coletadas. Analisando cada marcador biológico, concluímos que as populações estudadas não se tratam de complexo de espécies. Ainda não descartamos a existência de complexo dentro de Oc. scapularis, porém, para definitiva resposta a essa questão serão necessários mais estudos envolvendo populações de outras regiões. / Samples of Oc. scapularis were collected in the municipalities of Tremembé-SP (TRE), São Paulo-SP (SPA) and Itaboraí-RJ (ITA). We used the following biological markers: mitochondrial cytochrome oxidase subunit-1 gene (COI), wing geometry and shape of male genitalia. These markers are traditionally known by its discriminating power in studies of this nature. ITA, SPA and TRE populations, showed distinct wing shape, suggesting low gene flow. We observed sexual dimorphism concerning the isometric size, wing shape and the degree of populational differentiation. ITA sample exhibited the lowest centroid sizes. We found high genetic polymorphism in all populational samples, being 51 COI haplotypes. Out of them, only 11 haplotypes were noted to be shared between at least two or three populations. When comparing our results with those of a previous survey conducted by Devicari (2010), we found a quite similar pattern of hign polymorfism. In total, both studies comprised 52 haplotypes. The <font face=\"Symbol\">Fst index of genetic differentiation values were considered \"moderate\" between ITA and SPA and \"low\" in the other comparisons. Present results are consistent with the hypothesis that populations are subjected to isolation by geographical distance. Analyzing together each biologcal marker, we conclude that populations studied do not consist a species complex. We do not rule out the possible occurence of a complex in Oc. scapularis, however, a definitive answer to this question will require further studies and sampling of populations from elsewhere.

Submitted by (edna.saturno@ufrpe.br) on 2016-07-19T14:48:33Z No. of bitstreams: 1 Cristiane Maia da Silva.pdf: 1998511 bytes, checksum: 58b33e4aff3f932a9eed5c8296403044 (MD5) / Made available in DSpace on 2016-07-19T14:48:33Z (GMT). No. of bitstreams: 1 Cristiane Maia da Silva.pdf: 1998511 bytes, checksum: 58b33e4aff3f932a9eed5c8296403044 (MD5) Previous issue date: 2014-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The insects from culicidae family are present in different habitats since homes until places of habitation of domestics animals. The State of Pernambuco is considered endemic for various diseases transmitted by Culicidae. This is the first survey about culicids in the Municipality of Cabo de Santo Agostinho. The aim of this study was to evaluate the occurrence of different species of Culicidae in the Cabo de Santo Agostinho county located in Rainforest Zone of Pernambuco, Brazil. For this purpose, the larvae of culicidae from 3rd and 4th instars were collected weekly during the period from October 2012 to September 2013. Twenty-three artificial traps were distributed and installed in domiciliary environments, peridomiciliary and within the forest, 20 meters away between them. The larvae were collected and transported in plastic containers containing water. When the larvae arrived in the laboratory were identified. A total of 12,718 culicids larvae (average of 1059.83 specimens / month) were collected during study period. The largest number of specimens was detected in September 2013 (n = 4,084) and lowest in December 2012 (n = 86). Notably, 2,441 (19.2%), 7,098 (55.9%) and 3,179 (24.9%) larvae were collected in domiciliary, peridomiciliary environments and at forest, respectively. In relation to the placement of traps (peridomicile and forest) 8,585 (67.5%) specimens were collected on the floor and 1,692 (13.3%) in the treetop. Ten different species of Culicidae were identified, the more often was Aedes albopictus (46.45%; 5,908/12,718), Culex maxi (35.56%; 4,523/12,718) and Limatus durhami (12.58%; 1,600/12,718). Interestingly, Ae. albopictus was the only species detected throughout all the study period (average of 492 specimens / month). In distributed traps in domicile environments Ae. albopictus was responsible for 95.58% (2,333/2,441) of the samples, while the predominance in around peridomicile (floor) traps were Cx maxi (73.22%; 4,523/6,177) species and (treetop) Ae. albopictus (97.94%, 902/921). The presence of Oc. scapularis was exclusively observed in the peridomicile floor traps, demonstrating the preference of this species in this local. This is very worrying, because this specie is epidemiologically important as vector in filariasis and arboviruses transmission. Thus, there is a need of evaluate the vector competence of the Oc. scapularis population of Cabo de Santo Agostinho, Pernambuco for D. immitis, W. bancrofti and arboviruses, considering epidemiologics aspects and public health importance. Although Ae. albopictus, the most common specie in the studied area, has presented two population peaks from February to April and from June to September, it is also possible to conclude that the human and animal populations who live in studied area are exposed to culicids and pathogenics agents, especially, after the rains. Therefore, control and prevention measures of culicids should be adopted in the studied area in order to reduce the health impact of endemic species here reported. / Os culicídeos estão presentes em diferentes habitats frequentando domicílios humanos e locais de permanência de animais domésticos. O Estado de Pernambuco é considerado endêmico para diversas doenças transmitidas por culicídeos. Trata-se do primeiro levantamento de culicideos realizado na região do Cabo de Santo Agostinho, PE. O objetivo do presente estudo foi avaliar a ocorrência de diferentes espécies de culicídeos no município do Cabo de Santo Agostinho, na zona da Mata do estado de Pernambuco. Para tanto, larvas de culicídeos de 3º e 4º estádios foram coletadas, semanalmente, em criadouros artificiais no período de outubro de 2012 a setembro de 2013. Foram instaladas 23 armadilhas artificiais do tipo larvitrampa distribuídas em ambientes domiciliar, peridomiciliar e dentro da mata distando 20 metros entre elas. As larvas coletadas foram transportadas em recipientes plásticos contendo água e no laboratório foram identificadas. Um total de 12.718 larvas de culicídeos (média de 1.059,83 espécimes/mês) foi coletado durante o período de estudo. O maior número de espécimes foi detectado em Setembro de 2013 (n = 4.084) e o menor em Dezembro de 2012 (n = 86). Particularmente, 2.441 (19,2%), 7.098 (55,9%) e 3.179 (24,9%) larvas foram coletados nos ambientes domiciliar, peridomiciliar e de mata, respectivamente. Em relação ao posicionamento das armadilhas (peridomicílio e mata) 8.585 (67,5%) espécimes foram coletados no solo e 1.692 (13,3%) na copa. Dez diferentes espécies de culicídeos foram identificadas sendo Aedes albopictus (46,45%; 5.908/12.718), Culex maxi (35,56%; 4.523/12.718) e Limatus durhami (12,58%; 1.600/12.718) as mais frequentes. Curiosamente Ae. albopictus foi a única espécie detectada durante todo o período de estudo (média 492 espécimes/mês). Nas armadilhas distribuídas nos ambientes domiciliares houve predominância da espécie Ae. albopictus (95,58%; 2.333/2.441) enquanto nas posicionadas no peridomicílio (solo) predominou a espécie Cx. (Cx.) maxi (73,22%; 4.523/6.177) e (copa) Ae. albopictus (97,94%; 902/921). Nas armadilhas localizadas na mata várias espécies foram encontradas sendo Li. durhami (50,33%; 1.212/2.408) predominante no solo e Ae. albopictus no solo (38,12%; 918/2.408) e copa (55,71%; 430/771). Foi observada também a presença de Oc. scapularis, com frequência nas armadilhas instaladas no solo do peridomicílio demonstrando a domiciliação dessa espécie na área estudada, o que é muito preocupante, uma vez que essa espécie tem importância epidemiológica, pois possui competência vetorial para a transmissão de diversas arboviroses e filarioses. Conclui-se que há a necessidade, então, de avaliar o potencial vetorial da população de Oc. scapularis do Cabo de Santo Agostinho, PE para D. immitis, W. bancrofti e arboviroses considerando os aspectos epidemiológicos e a importância para saúde pública. Embora Ae. albopictus, a espécie mais frequente na área estudada, tenha apresentando dois picos populacionais, de Fevereiro a Abril e de Junho a Setembro, é possível concluir também que a população humana e animal residente na área de estudo está exposta aos culicídeos e aos agentes por eles veículados durante todo o ano, sobretudo após o período de chuvas. Portanto, medidas de controle e prevenção dos culicídeos devem ser adotadas na área estudada visando a redução do impacto sanitário causado pela endemicidade das espécies aqui reportadas.