Isolation and characterization of 15 microsatellite loci in the stingless bee Plebeia remota (Apidae: Meliponini)

The destruction of Brazilian natural habitats has reduced bee populations and negative impacts of native flora pollination have been noticed. This work describes the isolation and characterization of microsatellite loci and evaluates them as molecular markers to study genetic variability of the stingless bee Plebeia remota. A microsatellite enriched genomic library was constructed and 15 primer pairs were designed for this species. The survey was conducted by analyzing 21 unrelated individuals. Genetic diversity indexes were calculated. The mean allelic richness was 6.3, the observed heterozygosity was 0.568, and the percentage of polymorphic loci was 93.33%. Also the primers were tested in cross-species amplification and showed promising results for P. droryana, P. emerina, P. lucii, P. meridionalis, P. pugnax, and P. saiqui. The microsatellite loci described here will be useful to evaluate genetic variability of stingless bees, and certainly will improve our knowledge about population dynamics especially in threatened environments.

Morphometrical, biochemical and molecular tools for assessing biodiversity. An example in Plebeia remota (Holmberg, 1903) (Apidae, Meliponini)

We see today many efforts to quantify biodiversity in different biomes. It is very important then to develop and to apply other methodologies that allow us to assess biodiversity. Here we present an example of application of three tools with this goal. We analyzed two populations of Plebeia remota from two distinct biomes that already showed several differences in morphology and behavior. Based on these differences, it has been suggested that the populations of Cunha and Prudentopolis do not represent a single species. In order to verify the existence or absence of gene flow between these two groups, we characterized the patterns of mtDNA through RFLP, the patterns of wing venation through geometric morphometry, and the cuticular hydrocarbons through gas chromatography-mass spectrometry. We used bees collected in these two locations and also from colonies which have being kept for around 9 years at Sao Paulo University. We found six different haplotypes in these specimens, of which three of them occurred exclusively in the population of Cunha and three only in the Prudentopolis population. The fact that the populations do not share haplotypes suggests no maternal gene flow between them. The two populations were differentiated by the pattern of the wing veins. They also had different mixtures of cuticle hydrocarbons. Furthermore it was shown that the colonies kept at the university did not hybridize. These two groups may constitute different species. We also show here the importance of using other methodologies than traditional taxonomy to assess and understand biodiversity, especially in bees.

Molecular Characterization of Propolis-Induced Cell Death in Saccharomyces cerevisiae

Propolis, a natural product of plant resins, is used by the bees to seal holes in their honeycombs and protect the hive entrance. However, propolis has also been used in folk medicine for centuries. Here, we apply the power of Saccharomyces cerevisiae as a model organism for studies of genetics, cell biology, and genomics to determine how propolis affects fungi at the cellular level. Propolis is able to induce an apoptosis cell death response. However, increased exposure to propolis provides a corresponding increase in the necrosis response. We showed that cytochrome c but not endonuclease G (Nuc1p) is involved in propolis-mediated cell death in S. cerevisiae. We also observed that the metacaspase YCA1 gene is important for propolis-mediated cell death. To elucidate the gene functions that may be required for propolis sensitivity in eukaryotes, the full collection of about 4,800 haploid S. cerevisiae deletion strains was screened for propolis sensitivity. We were able to identify 138 deletion strains that have different degrees of propolis sensitivity compared to the corresponding wild-type strains. Systems biology revealed enrichment for genes involved in the mitochondrial electron transport chain, vacuolar acidification, negative regulation of transcription from RNA polymerase II promoter, regulation of macroautophagy associated with protein targeting to vacuoles, and cellular response to starvation. Validation studies indicated that propolis sensitivity is dependent on the mitochondrial function and that vacuolar acidification and autophagy are important for yeast cell death caused by propolis.

COMPOSITION OF FRESHLY HARVESTED BRAZILIAN ROYAL JELLY - IDENTIFICATION OF CARBOHYDRATES FROM THE SUGAR FRACTION

Samples of Brazilian royal jelly from Africanized Apis mellifera were analysed in order to determine the gross composition: crude moisture ranged from 67.80% to 69.40%, crude protein from 15.80% to 16.70%, crude lipid from 2.90% to 3.98% and-total sugars from 11.40% to 11.50%. The sugar fraction was investigated and revealed the presence of the following compounds identified by their retention time during HPLC analysis: ribose, fructose, glucose, sucrose, mannose, trehalose, erythritol, adonitol and mannitol.

Entomofauna visitante de Stenolobium stans (Juss.) Seem (Bignoniaceae), durante seu período de floração

Realizaram-se estudos sobre as formas de reprodução de Stenolobium stans (Juss.) Seem e determinou-se a diversidade, freqüência e constância dos insetos visitantes nas flores em diferentes horas, durante quatro anos. As flores de S. stans começam a se abrir nas primeiras horas do dia entre 5 e 6h, com duração de 3 a 8h. Quando o estigma está receptivo, o pólen tem 90% de viabilidade. Além do pólen, a flor possui outros atrativos para os insetos visitantes, ou seja, os osmóforos responsáveis pelo odor adocicado, luz ultravioleta refletida e néctar com 25% de açúcar. A planta é autocompatível, reproduzindo-se por autogamia, geitonogamia ou xenogamia o que determina a necessidade de polinizadores externos e justifica ser a espécie vegetal em estudo uma séria invasora de campos e pastagens. Grande diversidade de insetos foi verificada visitando as flores, com predominância das abelhas. Os polinizadores foram Centris collaris Lepeletier, Bombus morio (Swederus), Eulaema nigrita Lepeletier e Epicharis sp. No meio rural houve menor incidência das espécies nativas do que no ambiente urbano, com predominância da abelha introduzida Apis mellifera L. Fatores ambientais, principalmente a temperatura, luminosidade, umidade relativa do ar e velocidade do vento, influenciaram a atividade forrageadora dos insetos.

Visitantes florais de Lagerstroemia speciosa Pers: (Lythraceae)

Studies were carried out with Lagerstroemia speciosa Pers. on floral reproductive systems, diversity and constancy of visiting insects at different hours of day, the behaviour of these insects at the flowers and the influence of these environmental factors in relation to their visits. The fenology, anthesis and others particularity of this vegetal species was studied. A great diversity of insects was verified visiting the flowers with the predominance of bees. The most frequent and constant species encountered were: Nannotrigona testaceicornis (Lepeletier, 1836) (40,2%), Tetragonisca angustula (Latreille, 1811) (16,9%), Apis mellifera Linnaeus, 1758 (11,8%), Plebeia droryana (Friese, 1900) (9,1 %) e Exomalopsis fulvofasciata (Smith, 1879) (8,5%). The blossoms possessis features of melittophily syndrome and diurnal anthesis. The environmental factors influence the insects foraging activity, mainly temperature, light, time of day, humidity and wind speed. The effective pollinators were the large insects like Bombus morio (Swederus, 1787), Bombus atratus (Franklin, 1913), Centris tarsata (Smith, 1874), Centris flavifrons Fabricius, 1775, Xylocopa suspecta Camargo & Moure, 1988, Xylocopa frontalis (Olivier, 1789) and Eulaema nigrita Lepeletier, 1841.

Visitantes florais de Erythrina speciosa Andr. (Leguminosae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Entomofauna visitante das flores de Cassia spectabilis (L.) D C. (Leguminosae)

Realizaram-se estudos sobre sistemas de reprodução de Cassia spectabilis (L.) D C. (Leguminosae) observando-se a diversidade, freqüência e constância dos insetos visitantes em diferentes horários. Também testou-se a influência dos fatores ambientais em relação às visitas. Os resultados de polinização manual sugerem que C. spectabilis é autocompatível, porém, a xenogamia é o sistema de reprodução predominante. As inflorescências foram visitadas por uma grande quantidade de insetos, havendo predominância de abelhas. O horário de maior ocorrência dos insetos nas flores de C. spectabilis foi das 8 às 14 h e de menor ocorrência entre 7 e 8 h e das 17 às 18 h. Quanto ao comportamento dos insetos em relação à flor de C. spectabilis, observou-se que Xylocopa frontalis Olivier, X. suspecta Camargo & Moure, Bombus morio Swederus e Centris scopipes Friese possuem comportamento e morfologia adequados aos polinizadores legítimos; C. similis F., Oxaea flavescens Klug e Epicharis rustica flava Cockerell foram considerados polinizadores ocasionais. Pseudaugochloropsis graminea (F.), Tetragonisca angustula Latreille e A. mellifera L. foram considerados pilhadores. A polinização por vibração é o método usado pelas abelhas para coleta de pólen.

Trigona spinipes (Fabr.) (Hymenoptera: Apidae) em espécies de maracujazeiro: flutuação populacional, horário de visitação e danos às flores

Avaliou-se a infestação, flutuação populacional e horário de visitação de Trigona spinipes (Fabr.) (Hymenoptera: Apidae) em espécies de maracujazeiro. As espécies utilizadas foram Passiflora coccinea, P. setacea, P. alata, P. edulis f. flavicarpa, P. laurifolia e P. nitida, aos três anos de idade. O ensaio foi conduzido em parcelas de 1,5 m de comprimento, avaliando-se os dois lados da espaldeira, considerando-se apenas o 0,5 m superior desta e totalizando, para os dois lados, a área de 1,5 m². Avaliaram-se os seguintes parâmetros: número de irapuás e porcentagem de dano nos botões florais, flores, frutos, ramos, folhas e pedúnculos. Para a determinação do horário de visitação, foi utilizada P. coccinea, sendo as avaliações realizadas às 9:00h, 12:00h e 15:00h. A correlação entre o número médio de T. spinipes presentes nas flores de espécies de maracujazeiro e a porcentagem de danos foi positiva e significativa (r = 0,99). em nenhuma das observações efetuadas, constatou-se a presença de T. spinipes nos botões florais, frutos, ramos, folhas ou pedúnculos das espécies de maracujazeiro, não ocorrendo danos nessas estruturas. Esse resultado sugere que as abelhas são atraídas pelas flores ocasionando danos nessas estruturas, provavelmente, por utilizar o tecido floral ou resinas contidas neste para a construção de ninhos. P. coccinea foi a espécie mais danificada por T. spinipes, apresentando suscetibilidade ao ataque desse inseto. Os horários de maior incidência de T. spinipes foram 9:00h e 12:00h, ocorrendo decréscimo significativo no número de abelhas por flor às 15:00h. Constatou-se maior infestação de T. spinipes nas flores das plantas de maracujá nos meses de outubro a novembro, coincidindo com a primavera, em Jaboticabal, SP.

Otimização de superfícies seletivas de frequência com elementos pré-fractais utilizando rede neural MLP e algoritmos de busca populacional

This thesis describes design methodologies for frequency selective surfaces (FSSs) composed of periodic arrays of pre-fractals metallic patches on single-layer dielectrics (FR4, RT/duroid). Shapes presented by Sierpinski island and T fractal geometries are exploited to the simple design of efficient band-stop spatial filters with applications in the range of microwaves. Initial results are discussed in terms of the electromagnetic effect resulting from the variation of parameters such as, fractal iteration number (or fractal level), fractal iteration factor, and periodicity of FSS, depending on the used pre-fractal element (Sierpinski island or T fractal). The transmission properties of these proposed periodic arrays are investigated through simulations performed by Ansoft DesignerTM and Ansoft HFSSTM commercial softwares that run full-wave methods. To validate the employed methodology, FSS prototypes are selected for fabrication and measurement. The obtained results point to interesting features for FSS spatial filters: compactness, with high values of frequency compression factor; as well as stable frequency responses at oblique incidence of plane waves. This thesis also approaches, as it main focus, the application of an alternative electromagnetic (EM) optimization technique for analysis and synthesis of FSSs with fractal motifs. In application examples of this technique, Vicsek and Sierpinski pre-fractal elements are used in the optimal design of FSS structures. Based on computational intelligence tools, the proposed technique overcomes the high computational cost associated to the full-wave parametric analyzes. To this end, fast and accurate multilayer perceptron (MLP) neural network models are developed using different parameters as design input variables. These neural network models aim to calculate the cost function in the iterations of population-based search algorithms. Continuous genetic algorithm (GA), particle swarm optimization (PSO), and bees algorithm (BA) are used for FSSs optimization with specific resonant frequency and bandwidth. The performance of these algorithms is compared in terms of computational cost and numerical convergence. Consistent results can be verified by the excellent agreement obtained between simulations and measurements related to FSS prototypes built with a given fractal iteration

Agentes polinizadores e produção de grãos em cultura de café arábica cv. “Catuaí Vermelho”

Coffee (Coffea arabica L.) plants had the pollination process of their flowers studied in three coffee producing regions in Brazil: the first in Ribeirao Preto, state of São Paulo, in September of 2004, the second in Carmo do Paraiba, in the state of Minas Gerais, in October of 2006, and, the third, in Altinopolis, state of São Paulo, in September of 2007. In all these locations the coffee variety used was the 'Catuai Vermelho'. The pollinator species visiting the flowers, the frequency with which the flowers were visited by the insects, and the type of material collected by them ( nectar and/or pollen) were determined by counts taking place at the first 10 minutes of each hour from 7:00 AM till 6:00 PM during the months mentioned above. To estimate the fructification percentage, 25 branches were identified with color threads and protected with wire frames covered with a nylon fabric to prevent them from being visited by insects. Another group of 25 branches was similarly identified but remained without any covering so that their flowers were thus freely visited by insects. on the average, the most frequent pollinator was the Africanized bee Apis mellifera (73.7%) followed by the bee Trigona spinipes (14.5%), and Tetragonisca angustula (9.5%). The Africanized bee collected nectar and pollen to almost the same extent: 58.5% of the first and 41.5% of the second. To collect nectar, that species increased its visiting frequency up to 1:00 PM. To collect pollen, A. mellifera visited the flowers up to 1:00 PM with frequency peaks between 8:00 and 9:00 AM. T. spinipes collected pollen (58,5%) and nectar (41.5%). T. angustula collected only pollen. The branches freely visited by insects produced 181 grains on the average whereas those covered with the nylon fabric produced only 81 grains, this meaning a reduction of 55.25% in grain production. Coffee grains produced on the freely visited branches had a mean weight (1.13 g) significantly superior to that of those produced on the covered branches (0.94 g).

Environmental Fate of Neonicotinoids and Classification of Their Potential Risks to Hypogean, Epygean, and Surface Water Ecosystems in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Bee communities (Hymenoptera: Anthophila) of the Cerrado ecosystem in São Paulo State, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito Peltier em estruturas semicondutoras quasiperiódicas

There is nowadays a growing demand for located cooling and stabilization in optical and electronic devices, haul of portable systems of cooling that they allow a larger independence in several activities. The modules of thermoelectrical cooling are bombs of heat that use efect Peltier, that consists of the production of a temperature gradient when an electric current is applied to a thermoelectrical pair formed by two diferent drivers. That efect is part of a class of thermoelectrical efcts that it is typical of junctions among electric drivers. The modules are manufactured with semiconductors. The used is the bismuth telluride Bi2Te3, arranged in a periodic sequence. In this sense the idea appeared of doing an analysis of a system that obeys the sequence of Fibonacci. The sequence of Fibonacci has connections with the golden proportion, could be found in the reproductive study of the bees, in the behavior of the light and of the atoms, as well as in the growth of plants and in the study of galaxies, among many other applications. An apparatus unidimensional was set up with the objective of investigating the thermal behavior of a module that obeys it a rule of growth of the type Fibonacci. The results demonstrate that the modules that possess periodic arrangement are more eficient