Resumo: Zaprionus indianus (Diptera:Drosophilidae) é uma espécie provavelmente de origem africana e que rapidamente se dispersou por parte do continente sul-americano. Hoje, indivíduos dessa espécie são encontrados numa amplitude latitudinal de 35°, do Uruguai a Belém (Brasil). Em Z. indianus e nos demais insetos, as esterases constituem um grupo multifuncional e heterogêneo de enzimas que participam da hidrólise de ésteres, além de estarem relacionadas a diversos processos metabólicos. Neste estudo, foram realizadas análises de esterases de Z. indianus e Drosophila melanogaster, em géis de poliacrilamida (PAGE) a 10% de concentração, em indivíduos dos dois sexos e em diferentes fases do desenvolvimento. Os resultados indicaram que algumas carboxilesterases (respectivamente EST-2 e EST-5 de Z. indianus e EST-6 D. melanogaster) apresentam atividade acentuada em machos. Estas enzimas mostraram similaridades nas duas espécies, tais como o mesmo padrão de inibição, expressão principalmente no estágio adulto e aumento da atividade enzimática com o aumento da idade dos indivíduos. As similaridades bioquímicas entre as enzimas dos dois drosofilídeos sugerem ortologia entre seus genes codificadores. Foram então realizadas reações de PCR, utilizando oligonucleotídeos iniciadores desenhados com base na seqüência do gene Est-6 de D. melanogaster e o DNA genômico de Z. indianus. O fragmento amplificado foi seqüenciado, com composição GC de 48,5% e similaridade de 73% com a seqüência do gene de Est-6 de D. melanogaster. A análise da razão das substituições sinônimas e não sinônimas sugere uma proteína sob ação de seleção normalizadora, onde as trocas sinônimas são em sua maioria neutras e as não sinônimas, na maioria das vezes deletérias, foram eliminadas pela seleção natural. A modelagem ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Zaprionus indianus has expanded its geographical distribution since its recent invasion of the South American continent. The first record data of only eight years, and the origin is probably the South Africa. Nowadays, this species can be found in a latitudinal range of 35º, from Uruguay to Belem (Brazil). Esterases comprise a multi-functional and heterogeneous group of enzymes that participated in ester hydrolysis. In insects, they are related to several metabolic processes, including the reproductive function. Esterase patterns in Z. indianus and Drosophila melanogaster were characterized in polyacrylamide gels (PAGE). Two - esterases from Z. indianus, EST-2 and EST-5, showed similarity preference for substrate - naftil acetate and patterns of inhibition as compared to the EST-6 of D. melanogaster, suggesting a possible role in reproductive biology for both enzymes. Biochemical characterization of these esterases and its differential expression in males, suggest orthology among their genes. In this work, the genomic DNA from Z. indianus was submitted to PCR experiments using 3 sets of D. melanogaster Est-6 sequence primers. The PCR products were directly sequenced; its GC content was 48.5% and presented a 73% similarity compared to Est-6 D. melanogaster sequence. Analysis of sequence data, by estimates of nonsynonymous/synonymous rate ratios, showed that the majority of sites evolves under strong or moderate negative selection (81%) and a minority of sites (19%). is under significant positive selection. Molecular modeling indicates that the fundamental structural features important for catalysis are conserved in the esterase of Z. indianus and human bilesalt activated lipase (BAL), which was used as structural model. Structural and sequence comparisons suggest the evolutionary relationship between ...(Complete abstract click electronic access below) / Orientador: Carlos Roberto Ceron / Coorientador: Claudia Marcia Aparecida Carareto / Banca: Maura Helena Manfrin / Banca: Hermione de Campos Bicudo / Mestre

Made available in DSpace on 2014-06-12T15:05:47Z (GMT). No. of bitstreams: 2 arquivo4602_1.pdf: 941507 bytes, checksum: c0ae7f823963e2cbd711e367c4dd1d23 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2006 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Foram analisadas as linhagens Metahizium anisopliae var. acridum e Metarhizium anisopliae var. anisopliae quanto à patogênicidade sobre Zaprinus indianus, utilizando as concentrações 104, 105, 106, 107, 108 conídios/mL considerando o percentual de emergência de adultos. De acordo com a metodologia empregada verificou-se que as duas linhagens apresentaram ação contra Z. indianus. Os marcadores moleculares ITS (Internal Trancride Spacer) do rDNA, Intron Splice Site Primer e Microssatélite (SSR- Simple Sequence Repeats), foram utilizados para avaliar a diversidade genética entre as linhagens antes e após a passagem pela mosca. A análise de agrupamento usando o método de UPGMA baseada nas distâncias genéticas dos marcadores moleculares confirmou a diversidade genética reconhecida no gênero Metarhizium. O microssatélite (GTG)5 e o intron do grupo mRNA nuclear tiveram a mesma sensibilidade em detectar a variabilidade genética entre as linhagens de Metarhizium . Os produtos de amplificação dos loci ITS1-5.8-ITS2 do rDNA com os iniciadores ITS4 e ITS5 foram eficientes em demonstrar que as linhagens estudadas pertence à espécie Metarhizium anisopliae, apesar da diversidade genética demonstrada pelos marcadores (GTG)5 e EI1. Os perfis de amplificações da região microssatélite, intron e ITS após a passagem por Z. indianus comprovaram que as linhagens reisoladas foram às mesmas que foram utilizadas para infectar

Drosophila suzukii (Matsumura) (Diptera: Drosophila) is an invasive frugivore and has become a significant pest of small fruit, cherry and grape throughout the United States. It may be possible to determine if a Virginia vineyard is at risk of D. suzukii infestation by analyzing the biotic and abiotic factors around each vineyard. This pest is known to utilize a wide range of cultivated and wild host plants. A host plant survey was conducted at four vineyards in the Piedmont Region of Virginia to identify hosts used by D. suzukii around vineyards. The seasonal availability of host plants and adult emergence from them were tracked. Six host plant species of D. suzukii were identified, some available season-long. Monitoring D. suzukii in cultivated crops is crucial for the timing of spray applications. Homemade and commercially available baits and traps were deployed in two vineyards to determine the efficacy and selectivity towards D. suzukii. The homemade and commercially available baits that contained red wine caught the most D. suzukii, but none were exclusively attractive to D. suzukii. Wine grape susceptibility was assessed in laboratory choice and no-choice ovipositional bioassays. Ovipositional susceptibility was determined by measuring the physiological and morphological parameters using six wine grape varieties. More eggs were laid in grapes as penetration force decreased. Penetration force and not skin thickness was the limiting factor for oviposition. Survivorship of eggs laid in intact grapes was analyzed and survivorship to adulthood was dependent upon variety and survivorship usually exceeded 9% survival seen in previous studies. Larval developmental parameters of D. suzukii were affected by grape variety and the density of Z. indianus. D. suzukii mortality was increased in most cases when in competition with Z. indianus, but was less pronounced when reared in Viognier grapes. My Z. indianus oviposition study demonstrated that they will follow injury created by D. suzukii, and then the Z. indianus larvae may outcompete D. suzukii within the berries. These studies greatly improved our understanding of D. suzukii biology and ecology in Virginia vineyards. / Ph. D. / Drosophila suzukii (Matsumura) (Diptera: Drosophila), henceforth referred to as spotted wing drosophila (SWD) is an invasive pest of small fruits, cherries and grapes grown throughout the United States. This pest has a wide host range including cultivated and wild host plants. A host plant survey was conducted at four vineyards in the Piedmont Region of Virginia to identify those used by SWD. The seasonal availability of host plants and adult emergence from their fruit were tracked throughout the growing season. Six host plant species were found and these host plants were available to SWD throughout the season. Monitoring SWD in cultivated crops is crucial for the timing of spray applications. Homemade and commercially available baits and traps were deployed in two vineyards to determine the efficacy and selectivity towards SWD in the vineyard. Baits containing red wine, whether homemade or commercially available caught the most SWD, but none were exclusively attractive to SWD. Wine grape susceptibility to SWD oviposition was assessed in laboratory no-choice and choice ovipositional bioassays using six wine grape varieties; physiological and morphological parameters were considered. More eggs were laid in grapes as penetration force decreased. Penetration force and not skin thickness was the limiting factor for oviposition. Survivorship of eggs laid in intact grapes was analyzed and up to 50% of the eggs laid in larger grapes survived to adulthood. Larval interactions between SWD and Z. indianus, African fig fly (AFF), were also analyzed based on competition intensity and grape variety in which they were reared. SWD mortality, developmental parameters and pupal volume were impacted when in competition with AFF. SWD mortality was less pronounced, even when in competition with AFF, when reared in Viognier grapes compared to any other grape variety tested. My Z. indianus oviposition study demonstrated that they will flow injury created by D. suzukii and lay oviposit eggs into those wounds. These studies greatly improved our understanding of SWD biology and ecology in Virginia vineyards.

1.) Sentinel traps were used to survey for parasitoids of frugivorous drosophilids in Virginia fruit cropping systems, and determine if parasitoids were attacking invasive flies Drosophila suzukii (Matsumura) and Zaprionus indianus Gupta (Diptera: Drosophilidae) in the field. Two parasitoids of frugivorous drosophilids, Leptopilina boulardi (Barbotin, Carton, and Kelner-Pillault) (Hymenoptera: Figitidae) and Pachycrepoideus vindemiae (Rondani) (Hymenoptera: Pteromalidae), were reared, but only one P. vindemiae was reared from D. suzukii, and no parasitoids were reared from Z. indianus. Most parasitoids were reared from alternate host Drosophila melanogaster Meigen and other wild drosophilids. 2.) The ability of these parasitoids to attack D. melanogaster, D. suzukii and Z. indianus under controlled conditions was tested. Larval parasitoid L. boulardi did not develop on D. suzukii or Z. indianus, just D. melanogaster. Pupal parasitoid P. vindemiae successfully developed on all three fly species, but also increased pupal fly mortality. 3.) Olfactometry was used to ascertain if L. boulardi and P. vindemiae are selective about the type of fruit their hosts feed in. Results showed that among cherry, raspberry, blueberry, grape, and banana, L. boulardi preferred raspberry and banana to cherry, and preferred grape least, but no fruit was most preferred. Insufficient data were obtained for P. vindemiae. We conclude that parasitoids of Virginia are unlikely to provide effective biological control for D. suzukii or Z. indianus, and classical biological control should be investigated as a pest management option. Olfactometry results indicate tritrophic selectivity by Drosophila parasitoids, suggesting multiple parasitoids could be required for effective biological control. / Master of Science in Life Sciences / The spotted wing drosophila (henceforth SWD) is a globally invasive vinegar fly originating from southeast Asia, and is economically damaging to producers of small fruit, such as berries, cherries, and grapes. The African fig fly (henceforth AFF), a relative of SWD, is another recently invasive species to North America (originating from Africa) that often occurs simultaneously with SWD infestations, but its economic threat to North America is still unclear. With the economic threat posed by SWD, and potential threat posed by AFF, it is important to understand their relationships with other organisms in their environment, especially natural enemies, such as parasitoid wasps. Such information is integral for researchers to develop effective control methods, and will help determine if natural enemies can be used to our advantage as biological control agents. Biological control also helps to limit the use of chemical insecticides, mitigating the development of insecticide resistance in the pests. This project employed unique field trapping methods and laboratory bioassays to investigate the relationships of SWD and AFF with parasitoid wasps in affected fruit cropping systems in southwestern Virginia. We discovered that parasitoids of vinegar flies are present in Virginia fruit cropping systems, but they do not help to control populations of SWD and AFF. The parasitoids that are present prefer to attack other fly species, are unable to attack SWD and AFF, or do not attack in high enough numbers to have an impact on SWD or AFF populations. Biological control success is more likely to come from parasitoid species that have co-evolved with SWD and AFF in their native ranges.

Orientador: Carlos Roberto Ceron / Banca: Victor Hugo Valiati / Banca: Hermione Elly Melara de Campos Bicudo / Banca: Maria Tercília Vilela de Azeredo Oliveira / Banca: Hamilton Cabral / Resumo: Zaprionus indianus é uma espécie de mosca de origem africana que se dispersou pelas regiões tropicais da Ásia e das Américas. Grande interesse tem sido despertado sobre essa espécie, principalmente pela recente expansão da sua área de distribuição com a invasão do continente americano, ocorrida na década de 90. Hoje, indivíduos dessa espécie são encontrados em uma larga amplitude latitudinal, do Uruguai a Belém (Brasil), além do Panamá (América Central) e Flórida (Estados Unidos). Tem sido proposta que a introdução de Z. indianus no Brasil ocorreu no Estado de São Paulo e de lá a espécie se propagou para outros estados brasileiros por meio do comércio de frutas. Entretanto, a rota de dispersão dessa espécie não foi ainda completamente estabelecida. Na tentativa de esclarecer essa questão, avaliamos a variabilidade genética de dois marcadores, um fragmento do gene COI (citocromo oxidase I), de 612 pb, e do gene ortólogo ao Est-6 de Drosophila melanogaster, de 747 pb, em 19 populações de Z. indianus, de diversas regiões geográficas, englobando África, Ásia e Américas. A análise do polimorfismo do gene ortólogo ao Est-6 mostrou uma elevada diversidade nucleotídica, resultado de substituições sinônimas, indicando seleção purificadora atuando neste gene, tanto nas populações ancestrais como nas invasoras. Para o propósito de estabelecer a rota de dispersão de Z. indianus, foi construída uma rede de haplótipos por meio do método "median-joning network" para os dois marcadores. Os resultados são inéditos, pois indicam a ocorrência de duas invasões no Brasil, ao contrário de estudos anteriores que sugeriram apenas uma. Esses resultados também sugerem que a invasão de Z. indianus na América do Norte se deu a partir de migrantes de populações brasileiras, e não africanas ou asiáticas. Essa invasão pode estar diretamente... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Zaprionus indianus is a species of an African origin which has dispersed through tropical Asian as well as South and North America regions. Great interest has been taken at this species, mainly for the fact that recently its distribution has been expanded to American continent due to the invasion probably occurred in the nineties. Nowadays individuals from this species are found in wide latitudinal extent, from Uruguay to Belem (Brazil), besides Panama (Central America) and Florida (The United States). It has been proposed that Z. indianus introduction in Brazil happened from the São Paulo to other Brazilian states through fruit trading, though this species' dispersion route has not been completely established. In order to clarify this issue we evaluated genetic variability of two markers, a sequence of the gene COI (cytochrome oxidase I), 612 bp long and of the ortholog to Est-6 gene from Drosophila melanogaster, 747 bp long, in 19 populations of Z. indianus from several geographic regions, covering Africa, Asia and America. Polymorphism analysis of ortholog to Est-6 gene showed an elevated nucleotide diversity, resulting from synonym substitutions, indicating a purifying selection acting on this gene, which occurred in ancestral populations, as well as in the invader ones. Aiming at establishing the Z. indianus dispersion route, a haplotype network was built, using median-joining network method for the two markers. The results are unreleased, for they indicate the occurrence of two invasions in Brazil, in opposition to previous studies that have indicated just one. They also demonstrate that Z. indianus' invasion in North America has happened from Brazilian population migrants, not African or Asian ones. This invasion can be directly related to the fact that Brazil is currently the third largest world fruit producer and exports over... (Complete abstract click electronic access below) / Doutor