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.

Avaliação do serviço de polinização da abelha mandaçaia melipona quadrifasciata em macieiras.

O cultivo de macieira é uma atividade dependente da polinização cruzada para frutificação e, para tanto, da ação de visitantes florais para a transferência de pólen entre cultivares compatíveis (SALOMÉ; ORTH, 2014). O sucesso da polinização depende de uma série de fatores, entre os quais a eficiência do agente polinizador. A abelha Apis mellifera é a espécie mais comumente encontrada sobre flores de macieira. No entanto, não é a única espécie a realizar o serviço de polinização nos pomares. Segundo Lourdes et al. (1996) existe uma grande diversidade de espécies nativas de abelhas no interior de pomares de macieiras, as quais participam ativamente na polinização. A ação de polinizadores nativos é vista como complementar a realizada por A. mellifera (WITTER et al., 2014), e a descoberta de espécies potenciais para este fim é uma necessidade da pesquisa. Segundo Witter et al. (2014), a preocupação com o desparecimento de polinizadores pelo mundo, gerou inúmeros questionamentos sobre a dependência de uma única espécie de polinizador na agricultura. Embora escassas as informações sobre a utilização de abelhas sem ferrão em polinização de cultivos de importância econômica, há evidências do sucesso da ação de espécies nativas na 21 polinização de algumas culturas agrícolas. Por exemplo, a utilização da abelha mandaçaia Melipona quadrifasciata na polinização de cultivos de tomates possibilitou incremento no número de frutos por planta e melhora na qualidade dos frutos (SANTOS et al., 2009). Em macieira, uma alternativa seria o uso conjunto de abelhas nativas e africanizadas em serviços de polinização. De acordo com Viana et al. (2014) a introdução de colmeias de Melipona quadrifasciata anthidioides juntamente 26 com as de A. mellifera incrementou o número de sementes por fruto e a produção de frutos de macieira. Como a abelha mandaçaia Melipona quadrifasciata quadrifasciata apresenta maior atividade externa em horários de temperaturas mais baixas e de umidades relativas altas (GUIBU; IMPERATRIZ-FONSECA, 1984), diferentes aos apresentados por A. mellifera em macieira (SANTOS et al., 2013), pode haver menor competição entres as espécies pelo recurso floral, o que pode permitir ganhos na polinização de macieiras. Assim, objetivou-se avaliar a ação exclusiva da abelha mandaçaia M. quadrifasciata quadrifasciata na frutificação efetiva de macieiras cultivares Gala Baigent e Fuji Suprema.

A holistic approach for the investigation of bee health

Despite current evidence is in strong disagreement with an emergency for the conservation of Apis mellifera, great concern is related to profitability of beekeeping operations. A growing involvement of veterinary science in addressing bee health topics will therefore be fundamental to preserve and protect the entire sector. The experiments in this thesis focused on two different and interdependent levels related to bee health: the biochemical level and the parasitological level. At the biochemical level the impact of plant protection products on bee physiology and survival was studied, elucidating synergistic interactions between poor nutrition and pesticide exposure in A. mellifera and between an insecticide and a fungicide in Osmia bicornis. Moreover, an innovative fingerprinting approach on honey bee haemolymph was applied to detect population imbalances in the hive. The control of Varroa infestations was studied both at the biochemical and parasitological level. A panel of biomarkers in honey bee haemolymph was applied to compare different mite control protocols. This resulted in relevant indications for beekeeping operations pursuing the least impact on nutritional status of the colonies. To guide the decision making of beekeepers, a new formic acid evaporator was tested in comparison with a more established one. Considering its widespread distribution in the country, efforts were directed also towards N. ceranae. In particular, the pivotal aspect of diagnosis was studied, proposing a new qPCR method to overcome some limits of the existing ones. In conclusion, this works fills some of the knowledge gaps of the beekeeping sector. However, many of them still need to be addressed and the upcoming menaces of climate change and dispersal of pathogens via globalization should be targeted by research efforts in the near future. Therefore, a multifaceted vision of bee health is of capital importance, aware of the complementarity of reductionist and holistic approaches.

Physicochemical properties and antioxidant capacity of propolis of stingless bees (Meliponinae) and Apisfrom two regions of Tocantins, Brazil

ABSTRACTThe composition of propolis depends on time, vegetation and the location of the collection area. The objective of this study was to determine the physicochemical characteristics, the concentration of phenol compounds and the antioxidant capacity of propolis of native stingless bees (Meliponinae)and Apis from the State of Tocantins. Extraction with 80% ethanol (v/v) was performed in order to obtain the extracts. Parameters examined were: propolis mass loss by desiccation at 105 ºC, ashes, wax concentration and pH. Furthermore, the propolis antioxidant activity was measured, as well as the total concentration of phenol compounds. The extracts were also analyzed by high performance liquid chromatography. The total concentration of phenol compounds varied between 121.78 and 631.29 (mg GAE g-1). The antioxidant activity expressed by the value of CE50 varied between 29.81 and 845.38 µg mL-1. High performance liquid chromatography analysis allowed us to infer the existence of phenol compounds. The results indicated that the studied propolis samples constitute good sources of natural antioxidants. The variety of phenol compounds identified in this study, and the diverse biological functions reported in literature for these compounds indicated that this stingless bee propolis (Meliponinae) and Apis has a high pharmacological potential.

Sex determination in honey bees (Apinae and Meliponinae) and its consequences

The first experiments on sex determination in bees began with Dzierzon, Meves, Nachtsheim, Paulcke, Petrunkewitsch, Manning. Whiting, (1943) found multiple alleles in Bracon xo that are the Rosetta stone of sex determination in Hymenoptera. Whiting also discovered that some species of microhymenoptera do not possess xo sex alleles. Therefore, Hymenoptera apparently presents two types of sex determination superimposed on haplodiploidy. In the panmictic groups hemizygous (xo1, xo2,... xon) and homozygous (xo1xo1, xo2xo2... xonxon) are males while heterozygous (xo1xo2, ... xon-1xon) are females. There is no such series of xon in endogamous Hymenoptera, since the constant elimination of diploid males would be damaging to the population and the mutation of xo to xon would be quickly eliminated. Besides the Whiting hypothesis, four others are discussed. The new hypothesis of genomic imprinting, of Beukeboom, is eliminated since: a) spermatozoa that develop within the egg produce male tissue; b) telitokous parthenogenesis due to the fusion of two haploid cells develop into females; c) last instar larvae treated with juvenile hormone become queens. The Cunha and Kerr hypothesis (female determining genes are totally or partially additive and male determination is totally or partially nonadditive) explains all known cases. The xo is a female determining gene. Sex determination in social bees led to the gradual evolution of two systems of caste determination: one in which queens and workers are similar and males are very different (Apinae), and another in which workers and males are very similar and both very different from the queens (Meliponinae). This second system in stingless bees implies that many of the mutations that improve worker capacities also affect the males that will carry out some activities that in Apis are clearly female ones. Ten of these activities are described.

Lipid distribution in salivary glands of larvae and adult bees (Hymenoptera : Apidae)

Cytochemical studies were carried out to establish lipid distribution in the salivary glands of larvae and adult bees, using the imidazole buffer technique. In the duct cells of the larval salivary gland, the reaction was positive in the epicuticle and negative in the glandular lumen. The absence of smooth endoplasmic reticulum and the presence of lipids in the intercellular space suggest that lipids absorbed from the haemolymph could be used in the constitution of the epicuticle, after having been conveyed through the epithelium. In adult workers (new-emerged, nurse and forager workers), the head salivary glands presented a positive reaction in the secretion in glandular lumen, identifying its lipidic nature.

Presence of wall-devoid micro-organisms in hypopharyngeal glands of forager bees (Hymenoptera, Apidae)

The occurrence of mycoplasma-like bodies in the axial duct and intracellular canaliculli from hypopharyngeal glands of bees (Meliponinae and Apinae) is described. Since they are not found within cells and due to the absence of cellular alterations in the infected glands it is suggested that micro-organisms are not pathogenic to the bees.

Gender identification of five genera of stingless bees (Apidae, Meliponini) based on wing morphology

Currently, the identification of pollinators is a critical necessity of conservation programs. After it was found that features extracted from patterns of wing venation are sufficient to discriminate among insect species, various studies have focused on this structure. We examined wing venation patterns of males and workers of five stingless bee species in order to determine if there are differences between sexes and if these differences are greater within than between species. Geometric morphometric analyses were made of the forewings of males and workers of Nannotrigona testaceicornis, Melipona quadrifasciata, Frieseomelitta varia, and Scaptotrigona aff. depilis and Plebeia remota. The patterns of males and workers from the same species were more similar than the patterns of individuals of the same sex from different species, and the patterns of both males and workers, when analyzed alone, were sufficiently different to distinguish among these five species. This demonstrates that we can use this kind of analysis for the identification of stingless bee species and that the sex of the individual does not impede identification. Computer-assisted morphometric analysis of bee wing images can be a useful tool for biodiversity studies and conservation programs.

Differences between the quality of strawberries (Fragaria x ananassa) pollinated by the stingless bees Scaptotrigona aff. depilis and Nannotrigona testaceicornis

We investigated the success of two stingless bee species in pollinating strawberries in greenhouses. Three greenhouses and one open field area were used; one greenhouse had only strawberry plants (control), another (G1) had three colonies of Scaptotrigona aff. depilis and another (G2) had three colonies of Nannotrigona testaceicornis. In the open field area, the flowers could be visited by any bee. The total production of fruits was counted and a random sample (N = 100) from each area was used to measure weight, length, circumference, and achenes number (N = 5). The percentages of deformed strawberries were: 23% (no bees); 2% (greenhouses with bees) and 13% (open field). The strawberries from the greenhouse with N. testaceicornis and the open field were heavier than those from the greenhouses with no bees and with S. depilis. The fruit circumference was largest in the greenhouses with bees. The achenes number did not differ among the experimental areas. The strawberries produced in the greenhouses with stingless bees had more quality and greater commercial value than the fruits produced in the open field area and the greenhouse without bees. We conclude that stingless bees are efficient pollinators of strawberry flowers cultivated in greenhouses.

Trap-nesting bees (Hymenoptera: Apoidea) in forest fragments of the State of Sao Paulo, Brazil

We studied the community ecology of trap-nesting bees in two forest fragments of the State of Sao Paulo, Brazil, during two years, utilizing bamboo canes and tubes made of black cardboard as trap nests. The traps were inspected once a month with an otoscope. One hundred and fifteen nests were obtained at Estacao Ecologica de Paulo de Faria, Paulo de Faria (EEPF). These included nine species belonging to five genera and two families. At Santa Cecilia Farm (SCF), 12 species belonging to seven genera and three families built 392 nests. Natural enemies reared from nests of both areas included Hymenoptera, Diptera and Coleoptera. Species richness was similar between the areas but the communities differed considerably in species composition. The higher diversity found at EEPF was due to more even distribution of the species. No difference was observed between the numbers of nests built in each year in each area. Although the species richness was lower in the cool/dry season of both years at SCF, and in the first year at EEPF, the nesting frequencies did not differ between seasons for both the overall community but for each of the most abundant species. No annual fluctuation in the frequencies of nesting was observed. As temperature and precipitation were not found to be significantly different between the two years of study in each area, we concluded that climatic stability resulted in population stability.

The first record on Brazilian stingless bees published 450 years ago by Hans Staden

Only a few decades after 1492, when Christopher Columbus arrived on a Caribbean island and Pedro Alvares Cabral claimed Brazil for Portugal in 1500, a German mercenary gave the first description of stingless bees in 1557. He got to know them when he was imprisoned for months by an anthropophagous tribe in the coastal region of Santos, today in the State of Sao Paulo. This rather short but nevertheless extremely exact record on stingless bees is hidden in the first book on Brazil. Three species and important aspects of their life history were treated. This early description has been completely overlooked by bee scientists until now. My note intends to close this evident gap.

Male sleeping aggregations of solitary oil-collecting bees in Brazil (Centridini, Tapinotaspidini, and Tetrapediini; Hymenoptera: Apidae)

Males of solitary bees usually spend the night in clusters on small branches of plants, cavities and flowers. The individuals usually return to the same location each evening during their life, exhibiting site fidelity to a particular plant. We report on the sleeping roosts of the males of some oil-collecting bees of the genera Centris, Paratetrapedia, Lanthanomelissa, Monoeca, and Tetrapedia, as well as the host plants. We discuss the role of the male clusters to the associated plants.

Chemical analyses confirm a rare case of seed dispersal by bees

It has been suggested that dispersal of seeds of Coussapoa asperifolia magnifolia could have endozoochoric dispersal by frugivorous birds and monkeys because the fruits are red when ripe, or exozoochoric dispersal, because the exocarp is mucilaginous and sticky. However, our field observations showed only stingless bees collecting the exocarp with seeds of C. asperifolia magnifolia, which are used for building and repairing their nests, from which the plants sprout. This paper aimed to determine the fruit chemical composition, since we postulated that C. asperifolia magnifolia is neither consumed by birds nor monkeys due to being very sticky and apparently resinous. Apolar extract analyses revealed the fruits are not resinous but extremely rich in waxes ( mainly esterified triglycerides), and polar extract analyses revealed the sugar content to be close to the sensorial minimum level. This probably accounts for why only stingless bees are seen visiting fruits and dispersing seeds.

Pheromone paths attached to the substrate in meliponine bees: helpful but not obligatory for recruitment success

In contrast to marking of the location of resources or sexual partners using single-spot pheromone sources, pheromone paths attached to the substrate and assisting orientation are rarely found among flying organisms. However, they do exist in meliponine bees (Apidae, Apinae, Meliponini), commonly known as stingless bees, which represent a group of important pollinators in tropical forests. Worker bees of several Neotropical meliponine species, especially in the genus Scaptotrigona Moure 1942, deposit pheromone paths on substrates between highly profitable resources and their nest. In contrast to past results and claims, we find that these pheromone paths are not an indispensable condition for successful recruitment but rather a means to increase the success of recruiters in persuading their nestmates to forage food at a particular location. Our results are relevant to a speciation theory in scent path-laying meliponine bees, such as Scaptotrigona. In addition, the finding that pheromone path-laying bees are able to recruit to food locations even across barriers such as large bodies of water affects tropical pollination ecology and theories on the evolution of resource communication in insect societies with a flying worker caste.