Microsatellite loci for euglossine bees (Hymenoptera: Apidae)

The Neotropical Euglossini (Hymenoptera: Apidae) are important pollinators of many flowering plants, particularly orchids. Lack of highly polymorphic genetic markers for euglossine species has limited the study of their social organization and inbreeding. We therefore developed microsatellite markers for two species, Eulaema nigrita (11 loci) and Euglossa cordata (nine loci), most of which were highly polymorphic in the source species and in a range of related euglossine bees.     

Frugivory by a stingless bee (Hymenoptera: Apidae)

Frugivory is not frequent among bees. Although stingless bees visit aged fruits for pulp, the use of fresh fruits is recorded only for Trigona hypogea Silvestri, a species that does not visit flowers. Here we report the occurrence of frugivory in Trigona amazonensis (Ducke), a flower-visiting stingless bee.     

Ultrastructural characterization of spermatozoa in euglossine bees (Hymenoptera, Apidae, Apinae)

Summary. Euglossine spermatozoa are the longest described to date for the Hymenoptera. This cell includes a head and a flagellar region. In transverse sections, the acrosome is circular at the tip but has an oval contour along most of its length. The perforatorium penetrates into a deep cavity in the nuclear tip. The flagellum consists in an axoneme, a pair of mitochondrial derivatives, a centriolar adjunct and a pair of accessory bodies. The axoneme has a 9+9+2 microtubule pattern which becomes gradually disorganized in the final portion, with the central microtubules and the nine doublets terminating simultaneously, followed by the accessory microtubules. The mitochondrial derivatives are asymmetric both in length and diameter. Sectioned transversally, the derivatives are ellipsoidal or have a pear shape. The larger one has a more obvious paracrystalline region. The centriolar adjunct begins at the nuclear base and extends parallel to the axoneme until it encounters the smaller mitochondrial derivative, on which it fits, making a concave groove. In addition to these consistent euglossine features, species-specific differences that might be useful in phylogenetic work on the group are also noted.Received 18 October 2003; revised 4 September 2004; accepted 4 October 2004.     

Phylogeny,diversification patterns and historical biogeography of euglossine orchid bees (Hymenoptera: Apidae)

The orchid bees constitute a clade of prominent insect pollinators distributed throughout the Neotropical region. Males of all species collect fragrances from natural sources, including flowers, decaying vegetation and fungi, and store them in specialized leg pockets to later expose during courtship display. In addition, orchid bees provide pollination services to a diverse array of Neotropical angiosperms when foraging for food and nesting materials. However, despite their ecological importance, little is known about the evolutionary history of orchid bees. Here, we present a comprehensive molecular phylogenetic analysis based on ～4.0 kb of DNA from four loci [cytochrome oxidase (CO1), elongation factor 1‐α (EF1‐α), arginine kinase (ArgK) and RNA polymerase II (Pol‐II)] across the entire tribe Euglossini, including all five genera, eight subgenera and 126 of the approximately 200 known species. We investigated lineage diversification using fossil‐calibrated molecular clocks and the evolution of morphological traits using disparity‐through‐time plots. In addition, we inferred past biogeographical events by implementing model‐based likelihood methods. Our dataset supports a new view on generic relationships and indicates that the cleptoparasitic genus Exaerete is sister to the remaining orchid bee genera. Our divergence time estimates indicate that extant orchid bee lineages shared a most recent common ancestor at 27–42 Mya. In addition, our analysis of morphology shows that tongue length and body size experienced rapid disparity bursts that coincide with the origin of diverse genera (Euglossa and Eufriesea). Finally, our analysis of historical biogeography indicates that early diversification episodes shared a history on both sides of Mesoamerica, where orchid bees dispersed across the Caribbean, and through a Panamanian connection, thus reinforcing the hypothesis that recent geological events (e.g. the formation of the isthmus of Panama) contributed to the diversification of the rich Neotropical biota. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 552–572.     

The behavioural and structural aspects of grooming and related activities in euglossine bees (Hymenoptera: Apidae)

The movements and structures involved in grooming, resin and pollen manipulation, and collection of aromatic material are described for members of the tribe Euglossini. These bees typically groom with each leg while hanging by their mandibles, working from the head backwards. Collection and transfer of aromatic material is described after re-evaluation using fluorescent powder. Resin collection involves asynchronous ipsilateral transfer of the resin to the hindtibia. Pollen collection, in contrast, involves synchronous ipsilateral transfer. Areas of the body that are not groomed are illustrated, and the various plants and animals that exploit these regions discussed. The similarities between the behaviours of euglossines and bumblebees supports the idea that they are closely related.     

啶虫脒对西方蜜蜂蜂王培育质量的影响

【目的】本实验旨在研究王台中残留啶虫脒对西方蜜蜂Apis mellifera蜂王培育质量的影响。【方法】通过融化蜂蜡并添加啶虫脒药液制作王台,使各王台中分别含4个不同剂量的啶虫脒(0, 10, 100和1 000 μg/kg蜂蜡)。同时,控制蜂王产卵6 h, 3 d后,将孵化为1日龄的幼虫分别移入各组王台中,并放入蜂群哺育。移虫后第3和6天分别统计各组王台中幼虫的接受率和封盖率,待蜂王出房时,计算其出房率,测定蜂王个体初生重、胸重和胸宽指标;采用实时荧光定量PCR(qPCR)技术测定蜂王卵巢中卵黄原蛋白基因(Vg)、储存蛋白基因(hex110和hex70b)的相对表达量。【结果】100 μg/kg蜂蜡和1 000μg/kg蜂蜡这两个啶虫脒剂量组西方蜜蜂蜂王的出房率都显著低于0 μg/kg蜂蜡和10 μg/kg蜂蜡剂量组,而0 μg/kg蜂蜡与10 μg/kg蜂蜡剂量组之间及100 μg/kg蜂蜡与1 000 μg/kg蜂蜡剂量组之间出房率 均差异不显著;这4个剂量组的王台幼虫接受率和封盖率以及蜂王的初生重、胸重和胸宽均无显著差异。qPCR结果显示,Vg基因的相对表达量随啶虫脒浓度的增加而下降,其中,1 000 μg/kg蜂蜡剂量组Vg基因的相对表达量显著低于10 μg/kg蜂蜡剂量组和0 μg/kg蜂蜡剂量组,其余各剂量组之间差异不显著;这4个剂量组之间hex110和hex70b基因的表达量差异不显著。【结论】西方蜜蜂王台中啶虫脒残留超过100 μg/kg蜂蜡剂量时,会对蜂王培育产生不利影响。     

Microsatellite loci for the stingless bee Melipona rufiventris (Hymenoptera: Apidae)

Eight microsatellite primers were developed from ISSR (intersimple sequence repeats) markers for the stingless bee Melipona rufiventris. These primers were tested in 20 M. rufiventris workers, representing a single population from Minas Gerais state. The number of alleles per locus ranged from 2 to 5 (mean = 2.63) and the observed and expected heterozygosity values ranged from 0.00 to 0.44 (mean = 0.20) and from 0.05 to 0.68 (mean = 0.31), respectively. Several loci were also polymorphic in M. quadrifasciata, M. bicolor, M. mandacaia and Partamona helleri and should prove useful in population studies of other stingless bees.     

Bumble bee (Hymenoptera: Apidae) activity and pollination levels in commercial tomato greenhouses

Commercial greenhouse studies were conducted to assess levels of pollination of tomato (Lycopersicon esculentum Mill.) flowers in relation to bumble bee (Bombus impatiens Cresson) colony activity and colony densities. For the assessment of pollination levels of tomato flowers, five categories were defined based on bruising levels caused by bumble bee pollination. Colony activity was measured as bee trips per ha/d using electric powered photodiode monitors inserted into the hive entrance. Levels of pollination were positively correlated with bee activity levels, up to a mean of approximately 400 pollen grains per stigma per day, after which greater activity did not result in further increases in daily pollination levels. Densities of colonies in the commercial greenhouses studied ranged from 7.6 to 19.8 colonies per hectare with a mean of 11.6 +/- 0.9. We found that an average activity of 2,000 bee trips per hectare per day was more than adequate to ensure sufficient pollination, and that this level of activity could be achieved with 7-15 colonies per hectare, depending on greenhouse conditions. Greenhouses requiring >15 colonies per hectare to achieve this level of pollination may be able to increase bee activity through alteration of greenhouse conditions. Across 50-m rows of tomato plants, levels of pollination decreased with increasing distance from bee colonies, suggesting that colonies should be evenly distributed throughout the greenhouses.     

Effect of nectar composition and nectar concentration on honey bee (Hymenoptera: Apidae) visitations to hybrid onion flowers

Floral nectar characteristics of nine inbred lines of onion (Allium cepa L.) were examined to determine their influence on the attractiveness of the onion flowers to honey bees (Apis mellifera L.). Potassium concentrations and sugar concentrations of the nectar did not significantly correlate with the number of bee visits received by an umbel. The average amount of nectar produced by both the umbels and the individual florets was significantly positively correlated with the number of bee visits. Our results suggest that selection for flowers with high nectar production may lead to a higher rate of pollination of the onion seed crop.     

Survival of honey bee (Hymenoptera: Apidae) spermatozoa stored at above-freezing temperatures

The development of practical techniques for the storage of honey bee, Apis mellifera L., semen would significantly improve our ability to breed for desirable genotypes and maintain genetic diversity in populations. Artificial insemination of queens has been possible for some time, but the semen used is usually freshly collected, or held for < 1 wk at room temperature. I examined the limitations of spermatozoal survival at nonfrozen temperatures. Pooled, diluted semen was stored in sealed capillary tubes at room temperature (25 degrees C) or in a refrigerator set to 12 degrees C, for periods up to 1 yr. Survival of spermatozoa was assayed by a dual fluorescent staining technique using SYBR-14 and propidium iodide stains, which readily distinguishes live and dead cells. No significant loss of viable spermatozoa occurred within the first 6 wk. Between weeks 6 and 9, the percent live spermatozoa fell from 80 to 58%, and remained at that level until after 39 wk. By week 52, samples at room temperature, but not at 12 degrees C, fell to 18.9% live spermatozoa. Nonfrozen storage of honey bee semen has potential for short-term preservation of germplasm, however several factors need to be studied further to optimize survival rates.     

Long-term ecology of euglossine orchid-bees (Apidae: Euglossini) in Panama

Summary Abundance patterns during 6–7 years and orchid visitation were determined for 51 species of the 57 local euglossine bees. Male bees were counted at 3 chemical attractants presented in the same manner each month. Sites were separated by 75 km but included wet Atlantic forest at 500 m elevation, moist forest at 180 m near Barro Colorado Island, and cloud forest at 900 m near the Pacific ocean. 1. From 15 to 30 euglossine species of 4 genera were active in each month and site; monthly species number and general bee abundance were positively correlated. Many species had 3 annual abundance peaks (range 1–4) and were active throughout the year, but peak annual abundances rarely occurred during late wet or early dry seasons. In contrast, Eufriesea generally were present as adults only 1–2 months in a year. 2. Euglossine populations were exceptionally stable. Species at each site were more stable than any known insect population, and stability and abundance were positively associated. However, year-to-year population stability and the degree of seasonality were not correlated. Among the three sites, the more diverse (species rich) bee assemblages displayed lower stability; these were the wetter and upland sites. 3. The most abundant bees visited more orchid species. Eg. and El. each visited and average of 4 orchid species (range 0–13); Ex. and Ef. visited 0–3. Stable populations did not visit more or fewer orchid species than did unstable populations. 4. Less than 68% of species at each site visited orchid flowers; less than a few dozen of the 100–800 bees counted in a day carried orchid pollinaria. Over 20% of the euglossine species never were seen with pollinaria at any site and probably seldom visit orchids in central Panama. 5. Most bee species visited 1 or no fragrance orchids in a given habitat. Orchids tended to utilize common pollinators that seldom included more than 1 species, and they utilized stable or unstable, seasonal or aseasonal bees. However, the most stable and abundant bee, Eg. imperialis, rarely pollinated orchids; fewer than 10 of ca. 20000 bees carried pollinaria. 6. Orchids may interact primarily with discrete seasonal bee population peaks-probably the emerging adults. Although specialized orchid preferences are implicated for species that visit few or no local orchids but pollinate other species and carry pollinaria in other areas, euglossine bees do not need orchids to survive or reproduce.     

Influences on the density and dispersion of bumble bee nests (Hymenoptera: Apidae)

To determine some of the influences on nesting densities of bumble bees I examined the dispersion and occupancy of 35 nests in an old field on Amherst Island, Ontario, Canada and 39 nests found by Cumber (1953) in England. Densities up to 1 nest per 200 m² were observed. Overall, Cumber's nests were randomly dispersed, but nests on the ground surface were aggregated. Nests of surface-nesting species were consequently more aggregated than expected at random, whereas surface-nesting and underground-nesting species seldom nested near one another. This distribution of nests probably reflects the nesting habits of the rodents that originally constructed the nests, rather than interactions between bees. On Amherst Island, abnormally wet spring weather apparently hampered normal colony development, particularly of underground-nesting bees based on a low frequency of underground nests, scarcity of workers of two underground-nesting species compared with the relative abundance of spring queens, and delayed appearance of workers of all species relative to the previous year. The nests of two common species were randomly dispersed, but those of a third species were regularly distributed, suggesting that intraspecific interactions may limit nesting density for this species.     

Number of Malpighian Tubules in Larvae and Adults of Stingless Bees (Hymenoptera: Apidae) from Amazonia

The number of Malpighian tubules in larvae and adults of bees is variable. Larvae of Apis mellifera L. have four Malpighian tubules, while adults have 100 tubules. In stingless bees, this number varies from four to eight. The objectives of this study were to provide characteristics of the Malpighian tubules as well as to quantify their number in larvae and adults of six species of Meliponinae, Melipona seminigra merrillae Cockerell, Melipona compressipes manaosensis Schwarz, Melipona rufiventris Lepeletier, Scaptotrigona Moure, Frieseomelitta Ihering, and Trigona williana Friese. Malpighian tubules were dissected from larvae and adults, measured, quantified, and maintained in microtubes with Dietrich??s solution. The numbers of Malpighian tubules were constant only for larvae of M. rufiventris (four and eight) and Scaptotrigona sp. (four). The most frequent number of tubules in the Melipona group was seven and eight in larvae, and 70 and 90 in adults. In the Trigona group were four and 20 to 40, for larvae and adults, respectively. The results showed differences in the number of Malpighian tubules among the species analyzed and also between the larvae and adults of the same species. Despite the variation observed, species of the group Melipona always have a larger number and longer Malpighian tubules in both larvae and adults as compared to the Trigona group, which may indicate an evolutionary trend of differentiation between these groups.     

Bacterial probiotics induce an immune response in the honey bee (Hymenoptera: Apidae)

To explore immune system activation in the honey bee, Apis mellifera L., larvae of four ages were exposed through feeding to spores of a natural pathogen, Paenibacillus larvae larvae, to cells of a diverse set of related nonpathogenic bacteria, and to bacterial coat components. These larvae were then assayed for RNA levels of genes encoding two antibacterial peptides, abaecin and defensin. Larvae exposed to either P. l. larvae or a mix of nonpathogenic bacteria showed high RNA levels for the abaecin gene relative to controls. First instars responded significantly to the presence of the nonpathogenic mix within 12 h after exposure, a time when they remain highly susceptible to bacterial invasion. This response was sustained for two successive instars, eventually becoming 21-fold higher in larvae exposed to probiotic spores versus control larvae. The mixture of nonpathogenic bacteria is therefore presented as a potential surrogate for assaying the immune responses of different honey bee lineages. It also is proposed that nonpathogenic bacteria can be used as a probiotic to enhance honey bee immunity, helping bee larvae, and other life stages, survive attacks from pathogens in the field.     

Diversity in bee (Hymenoptera: Apoidea) and social wasp (Hymenoptera: Vespidae, Polistinae) community in "campos rupestres", Bahia, Brazil

Hymenoptera such as bees and social wasps are regular floral visitors in "campos rupestres" vegetation. A community of bees and social wasps was studied during floral visitation in an area of "campos rupestres", at Chapada Diamantina, BA, Brazil, from September 2001 to April 2002. The community was described in relation to diversity, evenness, and dominance rank, considering the individuals abundance (H' = 2.14/ J' = 0.55) and biomass (H' = 2.34/ J' = 0.60). Thirty nine bee (588 individuals/ 15.742 g) and 11 social wasp species (52 individuals/ 2.156 g) were collected, being the first report of social wasps for the Brazilian "campos rupestres". The main species regarding number of individuals were Trigona spinipes (Fabricius), Apis mellifera L., Frieseomelitta francoi (Moure), and Bombus brevivillus Franklin. About 48% of the species were represented by a single individual. There was an inversion in the dominance rank when the species biomass was considered. B. brevivillus, A. mellifera, T spinipes, and other species represented by 15 individuals or less, such as the social wasps Synoeca cyanea (Olivier), Polistes canadensis (L.) and Myschocyttarus drewseni (Saussure), and the bees Eufriesea nigrohirta (Friese), Xylocopa grisescens Lepeletier and Megachile (Pseudocentron) sp.l were the predominant species. The use of biomass in diversity analysis permitted to detect differences in the relative contribution of species in hierarchy dominance. The comparison between bee faunas from different areas indicates a large similarity of the sampled fauna in Palmeiras (Bahia State) with neighboring ecosystems, although with low values of similarity.