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.

Thermoregulation and Respiratory Metabolism in two Brazilian Stingless Bee Subspecies of Different Climatic Distribution, Tetragonisca angustula fiebrigi and T a angustula (Hymenoptera: Apidae: Meliponinae)

The thermoregulatory capacity of colonies of the stingless bee subspecies Tetragonisca angustula fiebrigi Schwarz 1938, and Tetragonisca angustula angustula Latreille 1807, was investigated during winter and summer. The temperatures [T] inside and outside the nests were measured for 48 hh every 2 hh. In the brood area, the mean T observed for T a fiebrigi are 28.1° and 29.5° C, respectively, during winter and summer, whereas for T a angustula they are 28.6° and 31.6° C The ambient T in the same period range from 10.5° - 24.4° C (winter) and 20.1 - 36.3° C (summer). In workers, the respiratory rates [RR] increase with a rise in T, however, the differences between workers of the subspecies are not significant in contrast to the RR measured within subspecies in winter and summer. The Q10 values indicate an optimal T range from 15 - 25° C in winter, and from 20 - 30° C in summer for T a fiebrigi. For T a angustula the corresponding values were 25 - 35° C and 30 - 40° C, respectively.

The role of wax and resin in the nestmate recognition system of a stingless bee, Tetragonisca angustula

Recent research has shown that entrance guards of the stingless bee Tetragonisca angustula make less errors in distinguishing nestmates from non-nestmates than all other bee species studied to date, but how they achieve this is unknown. We performed four experiments to investigate nestmate recognition by entrance guards in T. angustula. We first investigated the effect of colony odours on acceptance. Nestmates that acquired odour from non-nestmate workers were 63% more likely to be rejected while the acceptance rate of non-nestmates treated with nestmate odour increased by only 7%. We further hypothesised that guards standing on the wax entrance tube might use the tube as an odour referent. However, our findings showed that there was no difference in the acceptance of non-nestmates by guards standing on their own colony's entrance tube versus the non-nestmate's entrance tube. Moreover, treatment of bees with nestmate and non-nestmate resin or wax had a negative effect on acceptance rates of up to 65%, regardless of the origin of the wax or resin. The role of resin as a source of recognition cues was further investigated by unidirectionally transferring resin stores between colonies. Acceptance rates of nestmates declined by 37% for hives that donated resin, contrasting with resin donor hives where acceptance of non-nestmates increased by 21%. Overall, our results confirm the accuracy of nestmate recognition in T. angustula and reject the hypothesis that this high level of accuracy is due to the use of the wax entrance tubes as a referent for colony odour. Our findings also suggest that odours directly acquired from resin serve no primary function as nestmate recognition cues. The lack of consistency among colonies plus the complex results of the third and fourth experiments highlight the need for further research on the role of nest materials and cuticular profiles in understanding nestmate recognition in T. angustula.

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.

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).

Microdissection: a tool for bee chromosome studies

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

Bee Visitors of Quaresmeira Flowers (Tibouchina granulosa Cogn.) in the Region of Dourados (MS-Brasil)

This research was developed in the center-urban area at the city of Dourados (MS), in Tibouchina granulosa trees, a plant popularly known as quaresmeira in Brazil. The floral visitors of this species were recorded in three daily periods: 7 - 8 a.m., 1 - 2 p.m. and 4 - 5 p. m. Bee visitors of those plants were collected with an entomological net when they landed on the flowers. The bees were anesthetized in a closed camera containing etila acetate, conserved in Dietrich ' s fixative and, soon afterwards, transferred to 70% ethyl alcohol for subsequent identification. Almost 300 specimens of Africanized Apis mellifera, Trigona spinipes and Tetragonisca angustula were collected. The presence of these three species of bees in the flowers of T. granulosa suggests that they can be considered the main floral visitors of that vegetal species. T. angustula workers did not visit the flowers of T. granulosa between 7 and 8 a.m., but they were the main floral visitors from 4 to 5 p.m. The workers of T. spinipes presented a very aggressive behavior against the Africanized A. mellifera workers on the quaresmeira flowers, defending the food sources with strength and efficiency and provoking a drastic reduction in the number of honeybee visits between 1 and 2 p.m. Nevertheless, they did not interfere in the opportunistic activity of foraging by T. angustula workers, mainly between 4 and 5 p.m. The most important result of this research was the detection of the Competitive Exclusion Principle between A. mellifera and T. spinipes species in relation to the foraging behavior, when they exploit the floral resources of T. granulosa.

Entomofauna visitante de Belamcanda chinensis (L.) DC (Iridaceae) durante o período de floração

A study of the reproductive biology of B. chinensis (L.) DC. (Iridaceae) was realized comprising floral biology and breeding systems. The floral biology studies included analyses of nectar production, occurence of osmophores, corolla pigments, ultraviolet reflexion and absortion patterns, viability of pollen, pollinators and flower visitors. The breeding systems were studied taking into account the results of manual pollinators tests. B. chinensis is self-compatible bul cross-pollination is more frequent. The effective pollinators are Plebeia droryana (Friese, 1906) (45,7%), Trigona spinipes (Fabricius, 1793) (27,3%), Tetragonisca angustula (Latreille, 1811) (9,3%). Others insects visitors are considered nectar and pollen thieves. The flowering begins generally in January and February. The complete reproductive cicle, as here considered, begining with floral bud production ending with development of mature fruits, lasts January to June. Seed dispersion is ornitocoric.

Effect of Brazilian propolis on experimental hepatotoxicity in rats

Propolis (bee glue) is one of the major hive products of bees and is rich in flavonoids, which are known for their antioxidant activities. The aim of this study was to evaluate the hepatoprotective effects of the ethanolic extract of propolis (EEP) against experimental carbon tetrachloride (CCl 4)-induced liver toxicity in rats by means of biochemical indices. The animals were divided into 4 groups: GI= received mineral oil; GII= CCl 4(4mL/kg; Lp., single dose) treated; GIII= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Tetragonisca angustula, daily for 3 days and GIV= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Nannotrigonea testaceicornes, daily, for 3 days. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cholesterol and triacylglycerols were estimated after 3 days. CCl 4 caused a maximum increase (p<0,01) above biochemical parameters. As compared to CCl4 group (GII), the EEP (GIII and GIV) showed reduction in cholesterol, triacylglycerol, ALT, AST and alkaline phosphatase activity in the serum. In conclusion, these data indicate that EEP improved the dyslipidaemia, moreover, significantly attenuated increases in serum ALT and AST activities in rats with liver damage induced by carbon tetrachloride.

Características florais e dependência por polinizadores de cinco cultivares de pepino e manejo de colméias em estufas

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Enhanced production of parthenocarpic cucumbers pollinated with stingless bees and Africanized honey bees in greenhouses

Crops have different levels of dependence on pollinators; this holds true even for cultivars of the same species, as in the case of cucumber (Cucumis sativus). The aim of this research was to assess the attractiveness of flowers of three Japanese parthenocarpic cucumber cultivars and evaluate the importance of Africanized bees (Apis mellifera), and the Brazilian native stingless bees, Jata (Tetragonisca angustula) and Ira (Nannotrigona testaceicornis) on fruit production. Several parameters, including frequency of bee visits to flowers as well as duration of nectar collection and fruit set were examined; additionally, fruit weight, length and diameter were evaluated. Three greenhouses located in Ribeirao Preto, SP, were used for planting three cucumber cultivars (Hokushin, Yoshinari and Soudai). The female flowers were more attractive than male flowers; however, Jata bees were not observed visiting the flowers. The Africanized and the Ira bees collected only nectar, with a visitation peak between 10 and 12h. Visits to female flowers had a longer duration than visits to male flower visits in all three cultivars. Africanized bee colonies declined due to loss of bees while in the greenhouse; the native stingless bee colonies did not suffer these losses. When bees were excluded, fruit set was 78%; however, when bees had access to the flowers, fruit set was significantly (19.2%) higher. Fruit size and weight did not differ with and without bees. This demonstrates that even in parthenocarpic cucumber cultivars, which do not require pollination in order to from fruits, fruit production is significantly increased by bee pollination.

Similarity of heterochromatic regions in the stingless bees (Hymenoptera: Meliponini) revealed by chromosome painting

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

A morphologically specialized soldier caste improves colony defense in a neotropical eusocial bee

Division of labor among workers is common in insect societies and is thought to be important in their ecological success. In most species, division of labor is based on age (temporal castes), but workers in some ants and termites show morphological specialization for particular tasks (physical castes). Large-headed soldier ants and termites are well-known examples of this specialization. However, until now there has been no equivalent example of physical worker subcastes in social bees or wasps. Here we provide evidence for a physical soldier subcaste in a bee. In the neotropical stingless bee Tetragonisca angustula, nest defense is performed by two groups of guards, one hovering near the nest entrance and the other standing on the wax entrance tube. We show that both types of guards are 30% heavier than foragers and of different shape; foragers have relatively larger heads, whereas guards have larger legs. Low variation within each subcaste results in negligible size overlap between guards and foragers, further indicating that they are distinct physical castes. In addition, workers that remove garbage from the nest are of intermediate size, suggesting that they might represent another unrecognized caste. Guards or soldiers are reared in low but sufficient numbers (1-2% of emerging workers), considering that <1% usually perform this task. When challenged by the obligate robber bee Lestrimelitta limao, an important natural enemy, larger workers were able to fight for longer before being defeated by the much larger robber. This discovery opens up opportunities for the comparative study of physical castes in social insects, including the question of why soldiers appear to be so much rarer in bees than in ants or termites.