Resin-foraging by colonies of Trigona sapiens and T. hockingsi (Hymenoptera: Apidae, Meliponini) and consequent seed dispersal of Corymbia torelliana (Myrtaceae).

Resins are a critical resource for stingless bees and resin-collecting bees act as seed dispersers in tropical plants. We describe the diurnal foraging patterns of colonies of Trigona sapiens and T. hockingsi on resin and pollen. We also document patterns of waste removal and seed dispersal of Corymbia torelliana. At most, only 10% of foragers collected resin or dispersed seed. Nevertheless, bees dispersed 1-3 seeds outside the nest per 5 minutes, and 38-114 seeds per day for each nest. The proportion of returning bees carrying pollen was highest in the morning for both species. The proportion of foragers returning with resin loads showed no significant diurnal variation in any season. Waste removal activity peaked in the afternoon for T. sapiens and in the morning for T. hockingsi. Seed removal peaked in the afternoon in one year only for T. sapiens. Bees dispersed thousands of seeds of C. torelliana over the season even though only a small proportion of the colony was engaged in seed transport.

Host species of bees from the tribe Meliponini Nest in semi - arid Chaco forest near Las Lomitas, Formosa (Argentina)

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Host species of bees from the tribe Meliponini Nest in semi - arid Chaco forest near Las Lomitas, Formosa (Argentina)

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Morphological and molecular characters reveal differentiation in a Neotropical social bee, Melipona beecheii (Apidae : Meliponini)

Morphometrics and DNA microsatellites were used to analyse the genetic structure of populations of the stingless bee M. beecheii from two extremes of its geographic range. The results showed that populations from Costa Rica and Yucatan exhibit substantial phenotypic and molecular differentiation. Bees from Yucatan were smaller and paler than those from Costa Rica. The value of multilocus F-ST = 0.280 (P <0.001) confirmed that there were significant molecular genetic differences between the two populations. Populations showed significant deviation from Hardy Weinberg equilibrium and the values of FIS (the inbreeding coefficient) were positive for Costa Rica = 0.416 and the Yucatan Peninsula = 0.193, indicating a lack of heterozygotes in both populations possibly due to inbreeding. The DNA sequence of 678 bp of the mitochondrial gene COI differed between populations by 1.2%. The results of this study should be considered in conservation programmes, particularly with regard to the movement of colonies between regions.

Body size differs in workers produced across time and is associated with variation in the quantity and composition of larval food in Nannotrigona perilampoides (Hymenoptera, Meliponini)

Although variation in body size has been recently reported in stingless bees (Meliponini), empirical evidence evaluating possible factors related to such variation is lacking, and thus it is not clear if it may have an adaptive significance. We evaluated if variation in the body size and weight of workers of stingless bees fluctuates across a seasonal pattern and if this could be related to characteristics of the food consumed during the larval stage. The weight of larval provisions, their protein, and sugar content were evaluated in four colonies of Nannotrigona perilampoides every 2 months across 1 year. Worker-destined larvae from the same combs were allowed to develop and were sampled as callow workers to determine their weight and size using morphometric data. The weight and size of workers were highly correlated and varied across the seasons in established colonies, suggesting that size variation cycles across the year in stingless bees. An increase in the protein content and, to a lesser degree, the quantity of larval food were positively linked to variation in body weight and size; food with richer protein content resulted in larger and heavier workers. This study provides the first evidence of an effect of the quantity and composition of larval food on the size of workers in stingless bees. Although body weight and size of workers differed across seasons, they were not readily noticeable as changes seem to occur as a continuum across the year. Since size polymorphism was of a larger magnitude across time but not within age cohorts and as it was highly determined by food resources, it may not be an adaptive feature in stingless bees. However, more studies are needed to determine the role of the cyclical change in worker body size on colony performance and thus its adaptive significance in stingless bees.

Comparative study in stingless bees (Meliponini) demonstrates that nest entrance size predicts traffic and defensivity

Stingless bees (Meliponini) construct their own species-specific nest entrance. The size of this entrance is under conflicting selective pressures. Smaller entrances are easier to defend; however, a larger entrance accommodates heavier forager traffic. Using a comparative approach with 26 species of stingless bees, we show that species with greater foraging traffic have significantly larger entrances. Such a strong correlation between relative entrance area and traffic across the different species strongly suggests a trade-off between traffic and security. Additionally, we report on a significant trend for higher forager traffic to be associated with more guards and for those guards to be more aggressive. Finally, we discuss the nest entrance of Partamona, known in Brazil as boca de sapo, or toad mouth, which has a wide outer entrance but a narrow inner entrance. This extraordinary design allows these bees to finesse the defensivity/traffic trade-off.

Genetic structure of Partamona helleri (Apidae, Meliponini) from Neotropical Atlantic rainforest

Stingless bees of the genus Partamona are distributed from southern Mexico to southern Brazil. This genus has been subject to different approaches to solve questions concerning general biology, taxonomy, systematics and biogeography, but population studies applying molecular techniques are inexistent. We analyzed the genetic structure of P. helleri across its geographic distribution along the coastal Atlantic tropical rainforest in Brazil. Ten mtDNA haplotypes were observed in 47 colonies of P. helleri of which some were exclusive and others shared among geographic sub-groups. Statistical analysis showed high genetic differentiation between geographic areas sampled. Fragmentation of the Atlantic forest during Pleistocene glaciations is discussed as a possible cause of the present haplotype distribution and frequency.

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.

Characterization of microsatellite loci of Tetragonisca angustula (Hymenoptera, Apidae, Meliponini)

An enriched genomic library was constructed from Tetragonisca angustula, a stingless bee species widely distributed in Brazil. The library was screened using two simple-repeat oligonucleotide probes and 21 microsatellite primer pairs were designed flanking a selection of repeat sequences within positive clones. The polymorphism of the microsatellite loci was analyzed by screening a sample of 19 unrelated T. angustula workers. Fifteen out of 21 loci were shown to be polymorphic, with observed heterozygosity estimates ranging from 0.00 to 0.89. The primers were also successfully used to amplify microsatellite loci from other stingless bee species, Tetragonisca fiebrigi, Tetragonisca weyrauchi, Lestrimelitta maracaia and Schwarziana quadripunctata. The results from variability analyses suggest that the microsatellite loci isolated from T. angustula will be useful in further population studies for the species and also for other Meliponini.

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.

Ultrastructure of the ducts of the reproductive tract of males of Melipona bicolor bicolor Lepeletier (Hymenoptera, Apinae, Meliponini)

The present paper describes the ultrastructural features of seminal vesicle, post-vesicular vas deferens and ejaculatory duct of Melipona bicolor bicolor from newly emerged and mature males. Although the results do not show very consistent morphological signs of secretory activity by the epithelium of these organs, lipidic droplets and lamellar granules present in mature males' seminal vesicles and the vacuoles present in post-vesicular vas deferens are probably secretion. Besides, the spermatozoa in the lumen are immersed in a material of characteristic structure, which must be produced in superior regions of the reproductive system of immature males, not studied here. The presence of sperm cells, apparently in cytoplasm vesicles of seminal vesicle and post-vesicular vas deferens, suggests spermiophagy by their epithelium.

Class III glands in the abdomen of Meliponini

Class III tegumentar glands were studied in workers, as well as in queens and males when available, of 56 Meliponini species. The presence and development of these glands varies widely among and within species. However, the queen typically has more glands than do workers, and males rarely have any. Gland development in workers was evaluated by counting and determining the size of cells in histological sections. Laying queens were found to have more active gland cells than did virgins. Cell numbers and cell ultrastructure differed among glands similarly located in workers, queens and males. Cell size and ultrastructure also varied from tergite to tergite. In conclusion, since it is likely that most of them produce pheromones, the wide variability in these glands suggests that they are important to social interaction.

Comparative study of the ultrastructure and secretory dynamic of hypopharyngeal glands in queens, workers and males of Sceptotrigona postica latreille (Hymenoptera, Apinae, Meliponini)

The secretory cycle of hypopharyngeal glands (HPGs) in Scaptotrigona postica resembles that of Apis mellifera: in newly emerged workers the HPGs are in prefunctional state, their maximum development happens in the nurse workers and in forager workers they show signs of reabsorption. In S. postica these glands are also present in queens and males where they are more developed in newly emerged individuals. The ultrastructural features of the HPG secretory cycle in workers of S. postica and A. mellifera are alike: granular endoplasmic reticulum well developed, large secretion masses around the intracellular canaliculus in nurse workers and extensive degenerative structures in forager workers. Then it is suggested that the HPG secrete similar substances in both species. A second secretory cycle seems to occur in early foragers, may be with production of enzymes. The role of the HPGs in queens and males remains unknown but one possibility is enzyme production.

Tegumentary epithelial glands in the abdomen of virgin and physogastric queens of the stingless bee Scaptotrigona postica Latreille (Meliponini: Trigonina)

Nas operárias das abelhas eussociais, regiões do epitélio tegumentar do abdome podem hipertrofiar-se e tornarem-se glandulares, sendo responsáveis pela produção de cera usada na contrução dos alvéolos de cria. Nos meliponíneos, epitélio com essas características também foi observado nas rainhas, porém sua função ainda continua desconhecida. Teria esse epitélio função homóloga nas operárias? As rainhas, além da função reprodutiva, desempenhariam funções na colônia até então consideradas exclusivas das operárias? Para tentar colaborar no esclarecimento dessas questões realizou-se um estudo histológico e ultra-estrutural das glândulas epiteliais tegumentares do terceiro tergito em rainhas virgens e fisogástricas de Scaptotrigona postica Latreille. Os resultados morfológicos mostraram que o epitélio glandular existe e é facilmente discernível no terceiro tergito das rainhas, sendo muito mais desenvolvido nas fisogástricas do que nas virgens. A ultra-estrutura mostrou que há diferenças no tipo de organização do retículo endoplasmático liso entre as rainhas, o que, juntamente com os resultados da histologia e morfometria, indicam poder haver diferenças funcionais desse epitélio entre elas. Como já observado na literatura, nas rainhas virgens tal epitélio pode estar envolvido na produção de cera e confecção de alvéolos de cria, porém sua função nas rainhas fisogástricas é totalmente desconhecida. A hipótese lançada no presente estudo é de que esse epitélio pode estar envolvido na produção de feromônios que irão atuar nas interações sociais da rainha poedeira.