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.

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.

Composition and diversity of bees (Hymenoptera) attracted by Moericke traps in an agricultural area in Rio Claro, state of São Paulo, Brasil

Foram estudadas a composição e diversidade de abelhas em uma área agrícola no município de Rio Claro, Estado de São Paulo, de maio de 2003 a junho de 2004, utilizando armadilha de Moericke. O local de coleta, uma área com 58,08 hectares, caracteriza-se pela produção de grãos e a prática de plantio direto, sendo que 70% da área de entorno é utilizada para o plantio de cana-de-açúcar. Foram coletadas 456 abelhas distribuídas em 20 gêneros, pertencentes às famílias Andrenidae (4,8%), Apidae (40,8%) e Halictidae (54,4%). Espécimes dos gêneros Dialictus (38%) e Diadasia (30%) foram predominantes nesta área. A diversidade de espécies avaliadas pelos índices de Shannon e Simpson foram H =1,88 e 1/D= 4.15, respectivamente, e o índice de Equitatibilidade de 0,61.

Dwarf gynes in Nannotrigona testaceicornis (Apidae, Meliponinae, Trigonini). Behaviour, exocrine gland morphology and reproductive status

The behaviour and morphology of dwarf gynes produced in worker-sized cells of normal colonies in Nannotrigona testaceicornis (Meliponinae, Trigonini) were studied. The behaviour of these dwarf virgin queens was the same as observed for normal Trigonine gynes. The glandular equipment is also the same: Dufour glands, fat bodies and spermathecae are present. Despite these similarities, their ovaries are different. The functional significance of dwarf gynes is unknown, but may be a basis for an alternative reproductive strategy.

Hydrolases in the hypopharyngeal glands of workers of Scaptotrigona postica and Apis mellifera (Hymenoptera, Apinae)

Hydrolytic enzymes from hypopharyngeal gland extracts of newly emerged, nurse and foraging workers of two eusocial bees, Scaptotrigona postica, a native Brazilian stingless bee, and the Africanized honey bee (Apis mellifera) in Brazil, were compared. The hypopharyngeal gland is rich in enzymes in both species. Fifteen different enzymes were found in the extracts, with only a few quantitative differences between the species. Some of the enzymes present in the extracts may have intracellular functions, while others seem to be digestive enzymes. Scaptotrigona postica, had lower β-glucosidase and higher lipase esterase activities than A. mellifera. The differences may be due to different feeding habits and behavioral peculiarities of the two species. ©FUNPEC-RP.

Investigation of Partamona helleri (Apidae, Meliponini) B chromosome origin. An approach by microdissection and whole chromosome painting

The stingless bee Partamona helleri in southeast Brazil shows the regular chromosome number 2n = 34 and a variable number of up to four minute B1 or B2 chromosomes. Previous cytogenetic analyses have indicated morphological similarities between the B1 chromosome and chromosome segments in the regular karyotype. In this study, microdissection and chromosome painting were employed along with C banding, NOR banding, and base-specific fluorochrome staining to investigate the origin of the B1 chromosome in P. helleri. B1-generated probe hybridized exclusively to B1 chromosomes. This result suggests an independent origin from the regular karyotype or, alternatively, that the B chromosome may have suffered substantial genetic alterations along its independent evolution. The absence of higher dosages of these small B chromosomes in this population of P. helleri may be related to the existence of either a genetic or cytogenetic constraint in the establishment of such high numbered karyotypes. © 2012 INRA, DIB and Springer-Verlag, France.

Composição da fauna de Hymenoptera associada a área agrícola de manejo tradicional: abelhas nativas e parasitóides

Pós-graduação em Ciências Biológicas (Zoologia) - IBRC

Inventário de abelhas solitárias (Hymenoptera: Apoidea) com o auxílio de ninhos-armadilhas no campus da Unesp de Rio Claro - SP

As abelhas solitárias são caracterizadas pela independência da fêmea em construir e aprovisionar sozinha seus ninhos, sem auxílio, cooperação ou divisão de trabalho entre as fêmeas de uma mesma geração, ou entre mãe e filhas. Dentro deste grupo, se enquadram aquelas que nidificam em cavidades pré-existentes. O objetivo deste trabalho é inventariar as espécies de abelhas solitárias no Campus da UNESP de Rio Claro - SP com o auxílio de ninhos-armadilha. Para este estudo foram escolhidos quatro locais, e em cada um foram expostos 212 ninhos-armadilha, confeccionados com cartolina preta, fechados numa das extremidades, todos com o mesmo comprimento, porém diferentes quanto ao diâmetro: 12,0 x 0,6 cm (ninho pequeno, NP) e 12,0 x 0,8 cm (ninho grande, NG). Esses ninhos foram arranjados dentro de tubos de PVC e apoiados em prateleiras ou em troncos, sempre a 1,40 metros de altura e protegidos contra o sol e chuva. A amostragem dos ninhos foi realizada no período entre maio de 2009 e abril de 2010, com coletas a cada dez dias. Cada ninho retirado foi reposto para manter o número de ninhos disponíveis. O período de emergência foi acompanhado de maio de 2009 a agosto de 2010. Apenas um ponto de amostragem houve coletas de ninhos de abelhas, o “Sítio de Nidificação”, com nidificação de sete espécies de abelhas pertencentes às famílias Apidae, Centris (Hemisiella) tarsata, Centris (Heterocentris) analis, Tetrapedia curvitarsis e Tetrapedia diversipes, e Megachilidae, Carloticola sp., Epanthidium tigrinum e Megachile sp. A espécie mais abundante foi Tetrapedia curvitarsis, seguida por Centris (Heterocentris) analis e Tetrapedia diversipes. A construção de ninhos foi maior durante o período chuvoso, havendo no mês de fevereiro de 2010 maior abundância de nidificações (23%) assim como maior riqueza de espécies nidificando (n = 6). Neste mesmo mês também... (Resumo completo, clicar acesso eletrônico abaixo)

Inventário de vespas sociais (Hymenoptera, Vespidae) e comparação da eficiência de armadilhas atrativas em diferentes estratos vegetais na Floresta Estadual Edmundo Navarro de Andrade, Rio Claro, SP

Hymenoptera é uma das maiores ordens de insetos, com mais de 130 mil espécies descritas. Dentre as principais famílias destacam-se: Formicidae, Apidae e Vespidae, sendo que a última inclui a grande diversidade de vespas sociais presentes no Brasil. Este grupo tem grande importância no controle biológico de outras populações, como predadores ou controladores de pragas agrícolas, além de serem visitantes florais e polinizadores ocasionais. Sendo assim, o presente estudo consiste em um inventário das vespas sociais da Floresta Estadual Edmundo Navarro de Andrade (FEENA) em Rio Claro, SP, que possibilitou a comparação entre diferentes métodos de coleta empregados na captura de vespas sociais (coleta ativa e armadilhas atrativas em diferentes estratos na vegetação) e a comparação da fauna atual com um inventário realizado a mais de 30 anos na mesma localidade. Os dados foram coletados em diferentes estratos da vegetação utilizando armadilhas de garrafa PET contendo líquido atrativo (uma próximo ao solo, outra a 1,5 m de altura e a outra acima de 3 m na copa das árvores), e através de técnicas de coletas ativa com auxílio de redes entomológicas. Foram realizadas quatro coletas no decorrer de um ano, uma em cada estação. A riqueza das espécies do atual inventário foi comparada com a de Rodrigues & Machado (1982) e observou-se uma diminuição do número de espécies ao longo dos anos, de 32 para 21, sendo que 17 espécies foram amostradas em ambos os inventários. Dentre as armadilhas atrativas, houve diferença entre os estratos, sendo que as armadilhas de dossel mostraram ser de grande importância, pois capturaram 41,8% dos indivíduos e ainda apresentaram quatro espécies capturadas exclusivamente neste estrato. A técnica de coleta ativa com redes entomológicas também foi relevante, sendo responsável pela coleta de 93,75% dos indivíduos da tribo Mischocyttarini

Inventário de vespas sociais (Hymenoptera, Vespidae) e comparação da eficiência de armadilhas atrativas em diferentes estratos vegetais na Floresta Estadual Edmundo Navarro de Andrade, Rio Claro, SP

Hymenoptera é uma das maiores ordens de insetos, com mais de 130 mil espécies descritas. Dentre as principais famílias destacam-se: Formicidae, Apidae e Vespidae, sendo que a última inclui a grande diversidade de vespas sociais presentes no Brasil. Este grupo tem grande importância no controle biológico de outras populações, como predadores ou controladores de pragas agrícolas, além de serem visitantes florais e polinizadores ocasionais. Sendo assim, o presente estudo consiste em um inventário das vespas sociais da Floresta Estadual Edmundo Navarro de Andrade (FEENA) em Rio Claro, SP, que possibilitou a comparação entre diferentes métodos de coleta empregados na captura de vespas sociais (coleta ativa e armadilhas atrativas em diferentes estratos na vegetação) e a comparação da fauna atual com um inventário realizado a mais de 30 anos na mesma localidade. Os dados foram coletados em diferentes estratos da vegetação utilizando armadilhas de garrafa PET contendo líquido atrativo (uma próximo ao solo, outra a 1,5 m de altura e a outra acima de 3 m na copa das árvores), e através de técnicas de coletas ativa com auxílio de redes entomológicas. Foram realizadas quatro coletas no decorrer de um ano, uma em cada estação. A riqueza das espécies do atual inventário foi comparada com a de Rodrigues & Machado (1982) e observou-se uma diminuição do número de espécies ao longo dos anos, de 32 para 21, sendo que 17 espécies foram amostradas em ambos os inventários. Dentre as armadilhas atrativas, houve diferença entre os estratos, sendo que as armadilhas de dossel mostraram ser de grande importância, pois capturaram 41,8% dos indivíduos e ainda apresentaram quatro espécies capturadas exclusivamente neste estrato. A técnica de coleta ativa com redes entomológicas também foi relevante, sendo responsável pela coleta de 93,75% dos indivíduos da tribo Mischocyttarini

Techniques for the In Vitro Production of Queens in Stingless Bees (Apidae, Meliponini)

Considering the ecological importance of stingless bees as caretakers and pollinators of a variety of native plants makes it necessary to improve techniques which increase of colonies' number in order to preserve these species and the biodiversity associated with them. Thus, our aim was to develop a methodology of in vitro production of stingless bee queens by offering a large quantity of food to the larvae. Our methodology consisted of determining the amount of larval food needed for the development of the queens, collecting and storing the larval food, and feeding the food to the larvae in acrylic plates. We found that the total average amount of larval food in a worker bee cell of E varia is approximately 26.70 +/- 3.55 mu L. We observed that after the consumption of extra amounts of food (25, 30, 35 and 40 mu L) the larvae differentiate into queens (n = 98). Therefore, the average total volume of food needed for the differentiation of a young larva of F. varia queen is approximately 61.70 +/- 5.00 mu L. In other words; the larvae destined to become queens eat 2.31 times more food than the ones destined to become workers. We used the species Frieseomelitta varia as a model, however the methodology can be reproduced for all species of stingless bees whose mechanism of caste differentiation depends on the amount of food ingested by the larvae. Our results demonstrate the effectiveness of the in vitro technique developed herein, pointing to the possibility of its use as a tool to assist the production of queens on a large scale. This would allow for the artificial splitting of colonies and contribute to conservation efforts in native bees.

Microsatellite loci for the carpenter bee Xylocopa frontalis (Apidae, Xylocopini)

In the present study, a microsatellite-enriched genomic library was constructed and primers for 14 microsatellite loci were designed for Xylocopa frontalis. Twenty unrelated individuals were screened. All loci were polymorphic and the number of alleles per locus ranged from 6 to 17 (x = 10.43). Observed (H-o) and expected (H-e) heterozygosities ranged from 0.350 to 0.950 and 0.674 to 0.898, respectively. All loci were in Hardy-Weinberg equilibrium, except one. The microsatellite loci described in this study will contribute towards general biology studies of X. frontalis, intranidal genetic relationships and nest management for the pollination of passion fruit.

Mixed colonies of two species of congeneric stingless bees (Hymenoptera: Apinae, Meliponini) display environmentally-acquired and endogenously-produced recognition signals

Nestmate recognition is fundamental for the maintenance of social organization in insect nests. It is becoming well recognized that cuticle hydrocarbons mediate the recognition process, although the origin of recognition cues in stingless bees remains poorly explored. The present study investigates the effects of endogenously-produced and environmentally-acquired components in cuticular hydrocarbons in stingless bees. The tests are conducted using colonies of Plebeia droryana Friese and Plebeia remota Holmberg. Recognition tests are performed with four different groups: conspecific nestmates, conspecific non-nestmates, heterospecifics and conspecific, genetically-related individuals that emerge in a heterospecific nest. This last group is produced by introducing brood cells of P. droryana into a P. remota colony, and the resulting adult bees are tested for acceptance 10 days after emergence. For all groups, 15 individuals are sampled for chemical analysis. The results show the acceptance of all conspecific nestmates, and the rejection of almost every conspecific non-nestmate and every heterospecific bee. Genetically-related individuals emerging from heterospecific nests present intermediate rejection (66.7% rejection). Chemical analysis shows that P. droryana individuals emerging in a P. remota nest have small amounts of alkene and diene isomers found in P. remota cuticle that are not found in workers from the natal nest. The data clearly show that the majority of the compounds present in P. droryana cuticle are endogenously produced, although a few unsaturated compounds are acquired from the environment, increasing the chemical differences and, consequently, the rejection percentages.

The Ecological Basis for Biogeographic Classification: an Example in Orchid Bees (Apidae: Euglossini)

Biogeography has been difficult to apply as a methodological approach because organismic biology is incomplete at levels where the process of formulating comparisons and analogies is complex. The study of insect biogeography became necessary because insects possess numerous evolutionary traits and play an important role as pollinators. Among insects, the euglossine bees, or orchid bees, attract interest because the study of their biology allows us to explain important steps in the evolution of social behavior and many other adaptive tradeoffs. We analyzed the distribution of morphological characteristics in Colombian orchid bees from an ecological perspective. The aim of this study was to observe the distribution of these attributes on a regional basis. Data corresponding to Colombian euglossine species were ordered with a correspondence analysis and with subsequent hierarchical clustering. Later, and based on community proprieties, we compared the resulting hierarchical model with the collection localities to seek to identify a biogeographic classification pattern. From this analysis, we derived a model that classifies the territory of Colombia into 11 biogeographic units or natural clusters. Ecological assumptions in concordance with the derived classification levels suggest that species characteristics associated with flight performance, nectar uptake, and social behavior are the factors that served to produce the current geographical structure.

Cell-sealing efficiency and reproductive workers in the species Melipona bicolor (Hymenoptera, Meliponini): double standard and possible rogue conduct

In many hymenopteran insect societies, selfish workers are policed, as selfishness can negatively affect the average inclusive fitness of one or both castes by reducing either the degree of average relatedness to the colony's male offspring or colony efficiency. In stingless bees, the rapid capping of brood cells could aid in controlling selfishness; to this end, we studied cell-sealing efficacy in Melipona bicolor. Execution of cell sealing was found to be both rapid and almost continuous. Comparing the performance of reproductive and non-reproductive workers, the former sealed the cells more efficiently when they contained their own eggs, but less so when the queens' eggs were involved. We argue that the occurrence of disruptions in cell sealing through self-serving reproductive workers is capable of undermining sealing efficacy as a policing instrument, thus making reproductive workers potential rogue individuals.