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Perfume making by male orchid bees is a remarkable behavior giving rise to an entire pollination syndrome in the neotropics. Male orchid bees concoct and store perfume mixtures in specialized hind-leg pockets using volatiles acquired from multiple environmental sources, including orchids. However, the precise function and ultimate causes of this behavior have remained elusive. We supplemented maleEuglossa dilemmareared from trap-nests with perfume loads harvested from wild conspecifics. In dual-choice experiments, males supplemented with perfumes mated with more females, and sired more offspring, than untreated, equal-aged, control males. Our results demonstrate that male-acquired perfumes function as chemical signals emitted during courtship and received by females when selecting mates. Sexual selection might be a key agent shaping the evolution of perfume signaling.One-Sentence SummaryThe possession of exogenous volatiles increase male mating success and paternity in orchid bees.

Cuticular hydrocarbons (CHCs) are waxy compounds on the surface of insects that prevent desiccation and frequently serve as chemical signals mediating social and mating behaviors. Although their function in eusocial species has been heavily investigated, little is known about the evolution of CHC-based communication in species with simpler forms of social organization lacking specialized castes. Here we investigate factors shaping CHC variation in the orchid bee Euglossa dilemma, which forms casteless social groups of two to three individuals. We first assess geographic variation, examining CHC profiles of males and females from three populations. We also consider CHC variation in the sister species, Euglossa viridissima, which occurs sympatrically with one population of E. dilemma. Next, we consider variation associated with female behavioral phases, to test the hypothesis that CHCs reflect ovary size and social dominance. We uncover a striking CHC polymorphism in E. dilemma spanning populations. In addition, we identify a separate set of CHCs that correlate with ovary size, social dominance, and expression of genes associated with social behavior, suggesting that CHCs convey reproductive and social information in E. dilemma. Together, our results reveal complex patterns of variation in which a subset of CHCs reflect the social and reproductive status of nestmates.

Hematophagous arthropods often choose predictable feeding sites on their hosts´ body, presumably to maximize blood uptake while minimizing costs. Realized feeding sites can be host specific, mediated by intrinsic host characters or/and specific preferences of the blood feeder. We investigated feeding site specificity in a community of frog-biting midges (Corethrella spp.) and frog hosts in La Gamba, Costa Rica. Midge distribution on hosts differed significantly between 12 investigated frog species, indicating that intrinsic host properties influence potential feeding sites. However, realized feeding sites were also significantly different between four investigated Corethrella morphotypes, across all hosts but also within certain shared hosts, indicating feeding site partitioning among Corethrella. We propose that the diversity of feeding sites in Corethrella-frog associations is determined by an interaction of host traits, e. g. calling site, defensive behavior or skin thickness, and midge traits, especially body size and corresponding mouthpart size/structure.

Male neotropical orchid bees (Euglossini) collect volatiles from varied sources in their environment to compile complex, species-specific tibial perfumes, which are later released at mating sites. ...

Abundance and diversity of bumblebees have been declining over the past decades. To successfully conserve bumblebee populations, we need to understand how landscape characteristics affect the quantity and quality of floral resources collected by colonies and subsequently colony performance. We therefore investigated how amount and composition of pollen collected by buff-tailed bumblebee Bombus terrestris colonies was affected by the surrounding landscape (i.e. the proportion of forest, urban, semi-natural habitats) and how they were related to colony growth. Thirty B. terrestris colonies were placed at grassland sites differing in surrounding landscape. Colonies were established in spring when availability of flowering plants was highest, and their weight gain was monitored for 1 month. We additionally recorded the quantity and compared plant taxonomic composition and nutritional quality (i.e. amino acid composition) of pollen stored. Bumblebee colonies varied little in the pollen spectra collected despite differences in surrounding landscape composition. They collected on average 80 % of pollen from woody plants, with 34 % belonging to the genus Acer. Early colony growth positively correlated with total amount of woody pollen and protein collected and decreased with increasing proportions of semi-natural habitats and total amino acid concentrations. Our results suggest that woody plant species represent highly important pollen sources for the generalist forager B. terrestris early in the season. We further show that colony growth of B. terrestris is predominantly affected by the quantity, not quality, of forage, indicating that several abundant plant species flowering throughout the bumblebees’ foraging season may cover the colonies’ nutritional needs.

Insects rely on the olfactory system to detect a vast diversity of airborne molecules in their environment. Highly sensitive olfactory tuning is expected to evolve when detection of a particular chemical with great precision is required in the context of foraging and/or finding mates. Male neotropical orchid bees (Euglossini) collect odoriferous substances from multiple sources, store them in specialized tibial pouches and later expose them at display sites, presumably as mating signals to females. Previous analysis of tibial compounds among sympatric species revealed substantial chemical disparity in chemical composition among lineages with outstanding divergence between closely related species. Here, we tested whether specific perfume phenotypes coevolve with matching olfactory adaptations in male orchid bees to facilitate the location and harvest of species-specific perfume compounds. We conducted electroantennographic (EAG) measurements on males of 15 sympatric species in the genus Euglossa that were stimulated with 18 compounds present in variable proportions in male hind tibiae. Antennal response profiles were species-specific across all 15 species, but there was no conspicuous differentiation between closely related species. Instead, we found that the observed variation in EAG activity follows a Brownian motion model of trait evolution, where the probability of differentiation increases proportionally with lineage divergence time. However, we identified strong antennal responses for some chemicals that are present as major compounds in the perfume of the same species, thus suggesting that sensory specialization has occurred within multiple lineages. This sensory specialization was particularly apparent for semi-volatile molecules (‘base note’ compounds), thus supporting the idea that such compounds play an important role in chemical signaling of euglossine bees. Overall, our study found no close correspondence between antennal responses and behavioral preferences/tibial contents, but confirms the utility of EAG profiling for discovering certain behaviorally active compounds.

Speciation is facilitated by the evolution of reproductive barriers that prevent or reduce hybridization among diverging lineages. However, the genetic mechanisms that control the evolution of reproductive barriers remain elusive, particularly in natural populations. We identify a gene associated with divergence in chemical courtship signaling in a pair of nascent orchid bee lineages. Male orchid bees collect perfume compounds from flowers and other sources to subsequently expose during courtship display, thereby conveying information on species identity. We show that these two lineages exhibit differentiated perfume blends and that this change is associated with the rapid evolution of a single odorant receptor gene. Our study suggests that reproductive isolation evolved through divergence of a major barrier gene involved in chemically mediated pre-mating isolation via genetic coupling.

Male orchid bees (Euglossini) collect volatiles from their environment to concoct species-specific “perfumes”, which are later emitted at mating sites. Intensity, complexity or composition of perfumes may encode age (survival) of a male, but how the individual perfume develops over time needs to be clarified. We investigated chemical changes during storage in leg pockets. We injected a mixture of eight perfume compounds into pockets of Euglossa imperialis and only the two most volatile compounds decreased over 12 days. Using a different approach we found significant shifts in quantities of naturally occurring perfume compounds of Euglossa championi over 10 days, with the strongest decreases (up to 70% peak area) in highly volatile minor compounds, e.g. monoterpenes, and noteworthy increases (up to 40%) in some sesquiterpenoids. Corresponding shifts were observed in legs of dried bees, suggesting that no metabolic activity is required for the observed changes to occur. Our results confirm that male orchid bees are generally good at preserving collected perfumes. However, subtle shifts towards heavier compounds in blends may occur over the lifetime of individual bees, e.g. due to evaporation or in-pocket chemical reaction, with old males acquiring a more pronounced base note in their seasoned perfumes.

Male Neotropical orchid bees collect volatile chemicals from their environment and compose species-specific volatile signals, which are subsequently exposed during courtship display. These perfumes are hypothesized to serve as attractants and may play a role in female mate choice. Here, we investigated the potential of cuticular hydrocarbons as additional recognition cues. The cuticular hydrocarbons of males of 35 species belonging to four of the five extant euglossine bee genera consisted of aliphatic hydrocarbons ranging in chain lengths between 21 and 37 C-atoms in distinct compositions, especially between sympatric species of similar coloring and size, for all but one case. Cleptoparasitic Exaerete spp. had divergent profiles, with major compounds predominantly constituted by longer hydrocarbon chains (>30 C-atoms), which may represent an adaptation to the parasitic life history ("chemical insignificance"). Phylogenetic comparative analyses imply that the chemical profiles exhibited by Exaerete spp. are evolutionarily divergent from the rest of the group. Female hydrocarbon profiles were not identical to male profiles in the investigated species, with either partial or complete separation between sexes in multivariate analyses. Sexually dimorphic hydrocarbon profiles are assumed to be the basis for sex recognition in a number of insects, and thus may supplement the acquired perfume phenotypes in chemical information transfer. Overall, cuticular hydrocarbons meet the requirements to function as intraspecific and intersexual close range recognition signals; behavioral experiments are needed to determine their potential involvement in mate recognition.

Male neotropical orchid bees collect volatile substances from their environment and compose complex, species-specific odour blends in leg pouches. These perfumes are accumulated by the males over time and are exposed during pre-mating display. It has been hypothesized that perfumes are indicators of male genotypic quality and that females chose mates by the quality or intensity of their odour. Because direct experimental proof is lacking, we investigated whether the amount or complexity of male perfumes is related to male (1) size, (2) fluctuating (left–right) asymmetry, or (3) age, traits that are known to be sexually selected in other animals. We measured left and right forewings (cell node distances for 1 and 2, wing wear for 3) of Euglossa dilemma and Euglossa viridissima collected during dry and rainy seasons on the Yucatan peninsula, Mexico (total N = 768). Wing size was not related to the quantity or the complexity of perfume extracts in either species after excluding the effect of season, which positively affected both size and perfume load in E. dilemma. Wing asymmetry had also no effect, except in rainy season E. viridissima, where it was positively (not negatively) correlated with the quantity of perfume. Wing wear, an established age indicator of orchid bees, had the only consistent overall effect, being positively correlated with perfume amount and complexity in both species. This is in agreement with the idea that perfumes are an honest indicator of male survival capacity.

The mating system of European wool-carder bees, Anthidium manicatum, differs from that of most bees in three important aspects: females (1) are polyandrous and (2) mate continuously over the course of their reproductive life, while males (3) exhibit resource defence polygyny, that is, defend patches of food plants where copulations occur. To shed light on the evolution of this mating system we investigated male paternity using a combination of cage experiments and microsatellite genotyping of brood. We found that, although females possess a spermatheca for long-term sperm storage, most brood was fathered by males that had very recently mated with the breeding female, indicating pronounced last (or at least ‘late’) male sperm precedence. In the absence of males (male exclusion experiment) a large proportion of eggs remained unfertilized (resulting in haploid male offspring), but some diploid daughters arose from fathers that had been removed at least 11 days prior to egg laying. It appears that most A. manicatum eggs are fertilized with sperm from the bursa copulatrix, while the spermatheca serves only as a backup reservoir. This is the first demonstration of last male sperm precedence in aculeate Hymenoptera (bees, wasps, ants). We suggest that it has coevolved with ‘resource defence’ or

Social parasites can break into their host colonies using sensory deception, force, or both. To evaluate the role of sensory deception in eusocial obligate food robbers, we studied the Mesoamerican stingless bee Lestrimelitta niitkib –host species system, including preferred and nonpreferred host species. The use of citral as a propaganda substance is documented in L. niitkib , but possible mechanisms used by individual scouts to overcome host species recognition have not been studied. We analysed the cuticular profiles of L. niitkib and host species, coupled with bioassays of time to aggression (latency) and included data on host species raid frequency. We found that L. niitkib has a simple, but not insignificant, cuticular profile. Generally, L. niitkib cuticular profiles were similar to (but did not mimic) profiles of its preferred host species and differed from profiles of nonpreferred hosts. As expected, latency generally fitted a recognition system based on the degree of similarity between the cleptobiont's cuticular label and the host species template, with chemically similar species reacting slower and chemically distant species reacting rapidly to L. niitkib . There was a positive correlation between raid ratio and latency, indicating that the speed of detection and aggression towards L. niitkib scouts could influence host species selection. Cuticular profile similarity of individual L. niitkib scouts to host species may help L. niitkib scouts evade recognition and attacks from guards. In a further step, unnoticed L. niitkib scouts could successfully recruit nestmates to mass-raid host species colonies. The fact that L. niitkib can also plunder aggressive species, suggests that obligate cleptobiosis within its narrow biological niche could be characterized by flexibility in invasion strategies to allow exploiting a broad range of host species and be successful over evolutionary times.

Many insects rely on chemical signals to transmit precise information on the location, identity, and quality of potential mates. Chemical signals are often broadcasted at sites with physical properties that maximize signal propagation and signal transmission. Male neotropical orchid bees (Euglossini) perch and display on vertical branches and tree trunks in the forest to expose volatile blends (perfumes) that they previously collected from their environment. Previous studies have shown that the chemical composition of perfume blends is highly differentiated even between closely related species. However, variation in behavioral components of perfume exposure and male display remain poorly understood. We conducted a four-year study on orchid bee display sites (8 species) in pacific Costa Rica, using field observations along with chemical analysis and cage experiments to assess display niche partitioning among sympatric species. We evaluated the influence of physical factors (terrain, wind, light) on the distribution of perch sites and on display behavior, and tested a prediction of the sex pheromone-analogue hypothesis, i.e., that displaying males have above-average quantities or qualities of acquired perfumes. Males of different species displayed in the same general area and sometimes in close proximity to each other, but partitioned the display niche by selecting different perch diameters, display heights, and by displaying at different times of the day. Most perch sites were located inside the forest on elevated ground, especially along ridges, where stronger winds may help disperse perfume signals. Furthermore, the angular position of displaying males on perches was narrowly determined by wind direction, with males being positioned on the downwind side of the perch, where they would be most conspicuous to conspecifics approaching on an odor trail. Although our results generally support the hypothesis that perfumes serve as pheromone analogues, we did not find differences in the perfume composition of males caught at display perches and males captured at chemical baits. This suggests that, while chemical communication is an integral part of orchid bee display, male display activity is not determined by the history of, and success in, volatile acquisition. © 2017 by the Ecological Society of America Muchos insectos dependen de señales químicas para transmitir información precisa sobre la ubicación, identidad y calidad de posibles parejas. Las señales químicas a menudo se transmiten en sitios con propiedades físicas que maximizan la propagación y transmisión de señales. Los machos de abejas orquídeas neotropicales (Euglossini) se posan y exhiben en ramas verticales y troncos de árboles en el bosque para exponer mezclas volátiles (perfumes) que recolectaron previamente de su entorno. Estudios anteriores han demostrado que la composición química de las mezclas de perfumes está muy diferenciada incluso entre especies estrechamente relacionadas. Sin embargo, la variación en los componentes de comportamiento de la exposición al perfume y la exhibición masculina sigue siendo poco conocida. Realizamos un estudio de cuatro años sobre sitios de exhibición de abejas de orquídeas (8 especies) en el Pacífico de Costa Rica, utilizando observaciones de campo junto con análisis químicos y experimentos en jaulas para evaluar la partición de nichos de exhibición entre especies simpátricas. Evaluamos la influencia de factores físicos (terreno, viento, luz) en la distribución de los sitios de percha y en el comportamiento de exhibición, y probamos una predicción de la hipótesis del análogo de feromonas sexuales, es decir, que los machos exhibidores tienen cantidades o cualidades de perfumes adquiridos. Los machos de diferentes especies se exhibían en la misma área general y, a veces, muy cerca unos de otros, pero dividían el nicho de exhibición seleccionando diferentes diámetros de percha, alturas de exhibición y exhibiendo en diferentes momentos del día. La mayoría de los sitios de percha se ubicaron dentro del bosque en terrenos elevados, especialmente a lo largo de las crestas, donde los vientos más fuertes pueden ayudar a dispersar las señales de perfume. Además, la posición angular de los machos que se exhiben en las perchas se determinó estrechamente por la dirección del viento, con los machos colocados en el lado de la percha a favor del viento, donde serían más visibles para los conespecíficos que se acercan en un rastro de olor. Aunque nuestros resultados en general apoyan la hipótesis de que los perfumes sirven como análogos de feromonas, no encontramos diferencias en la composición del perfume de los machos capturados en las perchas de exhibición y los machos capturados en cebos químicos. Esto sugiere que, si bien la comunicación química es una parte integral de la exhibición de abejas orquídeas, la actividad de exhibición masculina no está determinada por la historia y el éxito en la adquisición de volátiles. © 2017 por la Sociedad Ecológica de América Muitos insetos dependem de sinais químicos para transmitir informações precisas sobre a localização, identidade e qualidade de parceiros em potencial. Os sinais químicos são frequentemente transmitidos em locais com propriedades físicas que maximizam a propagação e a transmissão do sinal. Abelhas machos da orquídea neotropical (Euglossini) se empoleiram e se exibem em galhos verticais e troncos de árvores na floresta para expor misturas voláteis (perfumes) que elas coletaram anteriormente de seu ambiente. Estudos anteriores mostraram que a composição química das misturas de perfume é altamente diferenciada, mesmo entre espécies estreitamente relacionadas. No entanto, a variação nos componentes comportamentais da exposição ao perfume e da exibição masculina permanece pouco compreendida. Conduzimos um estudo de quatro anos em locais de exibição de abelhas de orquídeas (8 espécies) no pacífico Costa Rica, usando observações de campo junto com análises químicas e experimentos em gaiolas para avaliar a partição de nicho de exibição entre espécies simpátricas. Avaliamos a influência de fatores físicos (terreno, vento, luz) na distribuição de locais de poleiro e no comportamento de exibição, e testamos uma previsão da hipótese análogo de feromônio sexual, ou seja, que os machos exibindo têm quantidades ou qualidades acima da média de perfumes adquiridos. Machos de diferentes espécies exibidos na mesma área geral e às vezes próximos uns dos outros, mas dividiram o nicho de exibição selecionando diferentes diâmetros de poleiro, alturas de exibição e exibindo em diferentes horas do dia. A maioria dos locais de poleiro ficava dentro da floresta em terreno elevado, especialmente ao longo de cumes, onde ventos mais fortes podem ajudar a dispersar os sinais de perfume. Além disso, a posição angular de exibição de machos em poleiros foi estritamente determinada pela direção do vento, com os machos sendo posicionados no lado do poleiro a favor do vento, onde seriam mais visíveis para co-específicos se aproximando em uma trilha de odores. Embora nossos resultados geralmente apoiem a hipótese de que os perfumes servem como análogos do feromônio, não encontramos diferenças na composição do perfume de machos capturados em poleiros de exibição e machos capturados em iscas químicas. Isso sugere que, embora a comunicação química seja parte integrante da exibição das abelhas das orquídeas, a atividade da exibição masculina não é determinada pela história e pelo sucesso na aquisição de voláteis. © 2017 pela Ecological Society of America Ruhr-University Bochum, Alemania Universidad Nacional, Costa Rica University of California, Estados Unidos Escuela de Ciencias Ambientales

International audience; AbstractThe abundance and the diversity of bumblebees have been declining for the past decades. While some species remain abundant and widespread, others have experienced vast declines. We tested if diet breadth and exclusiveness of floral resource use affected abundance and land-use response of bumblebees and honeybees in heterogeneous agricultural landscapes. Based on an ecological niche modelling approach, we discovered that three of 13 species (12 Bombus spp. and A. mellifera) responded negatively to land use, while no species responded positively. Overall abundance as well as land-use response of different bee species was unrelated to resource use. The generalist forager B. lapidarius, the slightly more specialised forager B. terrestris/lucorum species complex and the specialist forager B. wurflenii declined most with land use. Moreover, we found that honeybees and two bumblebee species contained higher levels of nitrogen than other non-predatory Hymenoptera and performed almost perfect homeostasis despite variable nitrogen contents of floral resources. We conclude that at least common species of social bees have good nitrogen homeostasis capacity. Generally, our data do not support the notion that species with more specialised floral resource use are more likely to suffer from land use.

Theory predicts that both stabilising selection and diversifying selection jointly contribute to the evolution of sexual signalling traits by (1) maintaining the integrity of communication signals within species and (2) promoting the diversification of traits among lineages. However, for many important signalling traits, little is known about whether these dynamics translate into predictable macroevolutionary signatures. Here, we test for macroevolutionary patterns consistent with sexual signalling theory in the perfume signals of neotropical orchid bees, a group well studied for their chemical sexual communication. Our results revealed both high species-specificity and elevated rates of evolution in perfume signals compared to nonsignalling traits. Perfume complexity was correlated with the number of congeners in a species’ range, suggesting that perfume evolution may be tied to the remarkably high number of orchid bee species coexisting together in some neotropical communities. Finally, sister-pair comparisons were consistent with both rapid divergence at speciation and character displacement upon secondary contact. Together, our results provide new insight into the macroevolution of sexual signalling in insects.

In orchid bees, males signal their availability as mates by fanning ‘perfumes’, i.e. blends of volatiles that are collected from environmental sources and stored in hind leg pouches. The chemical composition of such perfumes in males with either two or three mandibular teeth has previously led to the discovery of two sympatric, cryptic lineages within Euglossa viridissima Friese on the Yucatan peninsula, Mexico. Here, we combine chemical, morphological, and genetic data for an integrated characterization of the two lineages. The lectotype of E. viridissima Friese in the Museum of Natural History in Vienna has two mandibular teeth, and the species name viridissima must thus be assigned to the predominantly bidentate lineage, whereas the completely tridentate lineage is described as a novel species, Euglossa dilemma sp. nov. Bembe & Eltz. Chemical profiling and microsatellite genotyping revealed that E. viridissima males can occasionally (∼10% of individuals) express a third mandibular tooth, but this tooth is not positioned centrally on the mandible as in E. dilemma, but is displaced towards the tip. Thus, males of the two lineages can be unambiguously diagnosed by mandibular characters alone. Based on 889 bp of CO1 sequence data, we confirm that E. viridissima and E. dilemma constitute a monophyletic group within the genus Euglossa. However, CO1 alone failed to separate these two lineages due to the lack of parsimony-informative sites. Both species occur in broad sympatry across Central America, but the orchid bees recently introduced to Florida have three mandibular teeth in males, i.e. belong to E. dilemma. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 163, 1064–1076.

Habitat degradation and fragmentation are widespread phenomena in tropical regions. Negative effects on the biota are numerous, ranging from interruption of gene flow among populations, to the loss of genetic diversity within populations, to a decline in species richness over time. Orchid bees (Hymenoptera: Apidae: Euglossini) are of major conservation interest due to their function as pollinators of numerous orchid species and other tropical plants. Here, we used microsatellite markers to investigate the effects of geographic distance and habitat fragmentation on gene flow among populations. Populations of Euglossa dilemma in three geographic regions—the Yucatan peninsula (Mexico), Veracruz (Mexico), and Florida (USA)—were genetically structured predominantly across the regions, with the strength of differentiation among populations being positively correlated with geographic distance. Within geographic regions only little substructure was found, suggesting that dispersal is substantial in the absence of geographic or ecological barriers. In a second study, patterns of genetic differentiation among eight species of Euglossa were not related to habitat fragmentation following deforestation in southern Mexico (Veracruz). Specifically, most bee populations in the 9,800 ha forest remnant of Los Tuxtlas (Volcano San Martin) were neither differentiated from, nor had less genetic diversity than, populations in near-continuous forest separated from Los Tuxtlas by 130 km of agricultural land. Either occasional long distance dispersal across open areas has buffered the expected genetic effects of fragmentation, or the history of fragmentation in southern Mexico is too recent to have caused measurable shifts in allelic composition.

SUMMARY That hummingbird-pollinated plants predominantly have red flowers has been known for decades, but well-investigated research studies are still rare. Preference tests have shown that hummingbirds do not have an innate preference for red colours. In addition, hummingbirds do not depend solely upon red flowers, because white-flowered hummingbird-pollinated plants are also common and temporarily abundant. Here we show that both white and red hummingbird-pollinated flowers differ from bee-pollinated flowers in their reflection properties for ultraviolet (UV) light. Hummingbird-pollinated red flowers are on average less UV reflective, and white hummingbird-pollinated flowers are more UV reflective than the same coloured bee-pollinated ones. In preference tests with artificial flowers, neotropical orchid bees prefer red UV-reflecting artificial flowers and white UV-nonreflecting flowers over red and white flowers with the opposite UV properties. By contrast, hummingbirds showed no preference for any colour in the same tests. Plotting floral colours and test stimuli into the honeybees' perceptual colour space suggests that the less attractive colours are achromatic for bees and therefore more difficult to detect against the background. This underlying colour preference in bees might provide hummingbirds with a private niche that is not attractive to bees.

The measurement of insect visits to flowers is essential in basic and applied pollination ecology studies but often fraught with difficulty. Floral visitation is highly variable, and observational studies are limited in scope due to the considerable time necessary to acquire reliable data. The aim of our study was to investigate whether the analysis of hydrocarbon residues (footprints) deposited by insects during flower visits would allow reconstruction of the visitor community and prediction of seed set for large numbers of plants. In 3 consecutive years, we recorded bumblebee visitation to wild plants of comfrey, Symphytum officinale, and later used gas chromatography/mass spectrometry (GC/MS) to quantify bumblebee-derived unsaturated hydrocarbons (UHCs) extracted from flowers. We found that the UHCs washed from corollas were most similar to the tarsal UHC profile of the most abundant bumblebee species, Bombus pascuorum, in all 3 years. The species composition of the bumblebee communities estimated from UHCs on flowers were also similar to those actually observed. There was a significant positive correlation between the observed number of visits by each of three bumblebee species (contributing 3-68% of flower visits) and the estimated number of visits based on UHC profiles. Furthermore, significant correlations were obtained separately for workers and drones of two of the study species. Seed set of comfrey plants was positively correlated to overall bumblebee visitation and the total amount of UHCs on flowers, suggesting the potential for pollen limitation. We suggest that quantifying cumulative footprint hydrocarbons provides a novel way to assess floral visitation by insects and can be used to predict seed set in pollen-limited plants.

Male orchid bees collect volatiles, from both floral and non-floral sources, that they expose as pheromone analogues (perfumes) during courtship display. The chemical profile of these perfumes, which includes terpenes and aromatic compounds, is both species-specific and divergent among closely related lineages. Thus, fragrance composition is thought to play an important role in prezygotic reproductive isolation in euglossine bees. However, because orchid bees acquire fragrances entirely from exogenous sources, the chemical composition of male perfumes is prone to variation due to environmental heterogeneity across habitats. We used Gas Chromatography/Mass Spectrometry (GC/MS) to characterize the perfumes of 114 individuals of the green orchid bee (Euglossa aff. viridissima) sampled from five native populations in Mesoamerica and two naturalized populations in the southeastern United States. We recorded a total of 292 fragrance compounds from hind-leg extracts, and found that overall perfume composition was different for each population. We detected a pronounced chemical dissimilarity between native (Mesoamerica) and naturalized (U.S.) populations that was driven both by proportional differences of common compounds as well as the presence of a few chemicals unique to each population group. Despite these differences, our data also revealed remarkable qualitative consistency in the presence of several major fragrance compounds across distant populations from dissimilar habitats. In addition, we demonstrate that naturalized bees are attracted to and collect large quantities of triclopyr 2-butoxyethyl ester, the active ingredient of several commercially available herbicides. By comparing incidence values and consistency indices across populations, we identify putative functional compounds that may play an important role in courtship signaling in this species of orchid bee. Electronic supplementary material The online version of this article (doi:10.1007/s10886-010-9821-3) contains supplementary material, which is available to authorized users.

Male Neotropical orchid bees (Euglossini) collect volatile substances (fragrances) from flowers and other sources (e.g., decaying wood) and store them in specialized hind tibial pockets. The accumulated chemicals are later emitted during courtship display, presumably to lure conspecific females for mating. We analyzed tibial fragrances of males of 15 sympatric Panamanian species in the genus Euglossa to test whether communities of euglossine bees are chemically structured, and to elucidate whether male fragrance signals evolve to convey premating isolation. Our analysis revealed substantial chemical disparity among all lineages. Disparity was mediated by compounds that were exclusive to certain species but also by differences in relative quantity of shared compounds. We mapped tibial fragrance compounds present in each species on a DNA-based phylogeny (reconstructed using partial sequences of COI, EF1-alpha, ArgK, and Pol-II) and found that most dominant compounds were highly homoplasious. In an analysis of chemical differentiation in relation to phylogenetic divergence through time, disparity was greater than expected from a null model at any point during evolutionary history, suggesting that diversifying selection has shaped fragrance phenotypes. Notably, chemical disparity was greater within recently diverged lineages than among them, suggesting that chemical preferences in orchid bees evolved rapidly in the early stages of species divergence. We postulate communication interference as the possible mechanism behind the observed fragrance differentiation, which may be the product of reproductive character (fragrance) displacement. Our findings are consistent with the hypothesis that male fragrance signals evolve to convey premating isolation.

Neotropical orchid bees (Euglossini) are conspicuously different from other corbiculate bees (Apinae) in their lack of advanced sociality and in male use of acquired odors (fragrances) as pheromone-analogues. In both contexts, orchid bee mating systems, in particular the number of males a female mates with, are of great interest but are currently unknown. To assess female mating frequency in the genus Euglossa, we obtained nests from three species in Mexico and Panama and genotyped mothers and their brood at microsatellite DNA loci. In 26 out of 29 nests, genotypes of female brood were fully consistent with being descended from a singly mated mother. In nests with more than one adult female present, those adult females were frequently related, with genotypes being consistent with full sister–sister (r = 0.75) or mother–daughter (r = 0.5) relationships. Thus, our genetic data support the notions of female philopatry and nest-reuse in the genus Euglossa. Theoretically, single mating should promote the evolution of eusociality by maximizing the relatedness among individuals in a nest. However, in Euglossini this genetic incentive has not led to the formation of eusocial colonies as in other corbiculate bees, presumably due to differing ecological or physiological selective regimes. Finally, monandry in orchid bees is in agreement with the theory that females select a single best mate based on the male fragrance phenotype, which may contain information on male age, cognitive ability, and competitive strength.

Oil palm plantations today cover large areas of former tropical lowland rain forest in Southeast Asia and are rapidly expanding on the island of Borneo. Study of the community of ground-dwelling ants in different plantations in Sabah, Malaysia, over 2 years using tuna baiting, revealed that the oil palm plantation ground ant community was severely reduced in species richness in comparison to the forest interior, regardless of age, undergrowth cover, or proximity to neighbouring forest. The results indicate that oil palm plantation habitats, now covering more than 15% of Sabah’s land area, can sustain only about 5% of the ground-dwelling ant species of the forest interior. Nine of the 23 ant species baited in the plantations were never recorded inside forest. All numerically dominant ants were non-forest species. The most common species was Anoplolepis gracilipes, an invasive species present at 70% of all bait sites and known to cause ‘ecological meltdowns’ in other situations. The low frequency and species number of forest ground ants indicates that oil palm plantations act as effective dispersal barriers leading to community isolation in rain forest remnants. The replacement of natural forests with oil palm plantations poses a serious threat to the conservation of biodiversity on Borneo if similar results are confirmed in other taxa.

Summary Saltational changes may underlie the diversification of pheromone communication systems in insects, which are normally under stabilizing selection favoring high specificity in signals and signal perception [1–4]. In orchid bees (Euglossini), the production of male signals depends on the sense of smell: males collect complex blends of volatiles (perfumes) from their environment [5, 6], which are later emitted as pheromone analogs at mating sites [7]. We analyzed the behavioral and antennal response to perfume components in two male morphotypes of Euglossa cf. viridissima from Mexico, which differ in the number of mandibular teeth. Tridentate males collected 2-hydroxy-6-nona-1,3-dienyl-benzaldehyde (HNDB) as the dominant component of their perfume. In bidentate males, blends were broadly similar but lacked HNDB. Population genetic analysis revealed that tri- and bidentate males belong to two reproductively isolated lineages. Electroantennogram tests (EAG and GC-EAD) showed substantially lower antennal responses to HNDB in bidentate versus tridentate males, revealing for the first time a mechanism by which closely related species acquire different chemical compounds from their habitat. The component-specific differences in perfume perception and collection in males of two sibling species are in agreement with a saltational, olfaction-driven mode of signal perfume evolution. However, the response of females to the diverged signals remains unknown.

We compared nectar, pollen and resin loads of individual workers among colonies from six Trigona species in Sabah, Borneo. Individual bees rarely collected large amounts of both nectar and pollen during the same foraging trip. Instead, comparison of crop contents across departing, flower-visiting, and returning bees suggests that pollen-collecting workers often carried highly concentrated nectar in their crop upon nest departure. During their foraging trip, this crop nectar volume decreased progressively until crops were largely empty when they returned to their nest. Individually marked pollen foragers carried highly concentrated nectar when they left their nest, while crops and corbiculae from marked nectar foragers were empty upon departure. We suggest that a large proportion of previously stored and highly concentrated nectar may be required for pollen adhesion to corbiculae and/or serve as fuel during foraging on nectar-poor flowers.

International audience; Abstract Males of sibling orchid bees Euglossa viridissima and Euglossa dilemma are morphologically cryptic, except for the number and shape of mandibular teeth. An alternative morph of E. viridissima has a third tooth similar to males of E. dilemma. We used this model system to evaluate the potential of wing morphometrics for the resolution of these groups. We found differences in the size characters of forewings of E. viridissima and E. dilemma albeit with substantial overlapping amongst them. However, geometric morphometrics of forewing vein intersections separated both species and, to a lesser extent, morphotypes. A discriminant analysis of the shape of the radial cell showed separation between all three groups, too, albeit with higher misclassification between E. viridissima and E. dilemma. We show that sibling cryptic species and morphotypes can be identified by geometric morphometrics, supporting its application with other methods as powerful aids to infrageneric taxonomy in bees.

Background Insects rely more on chemical signals (semiochemicals) than on any other sensory modality to find, identify, and choose mates. In most insects, pheromone production is typically regulated through biosynthetic pathways, whereas pheromone sensory detection is controlled by the olfactory system. Orchid bees are exceptional in that their semiochemicals are not produced metabolically, but instead male bees collect odoriferous compounds (perfumes) from the environment and store them in specialized hind-leg pockets to subsequently expose during courtship display. Thus, the olfactory sensory system of orchid bees simultaneously controls male perfume traits (sender components) and female preferences (receiver components). This functional linkage increases the opportunities for parallel evolution of male traits and female preferences, particularly in response to genetic changes of chemosensory detection (e.g. Odorant Receptor genes). To identify whether shifts in pheromone composition among related lineages of orchid bees are associated with divergence in chemosensory genes of the olfactory periphery, we searched for patterns of divergent selection across the antennal transcriptomes of two recently diverged sibling species Euglossa dilemma and E. viridissima. Results We identified 3185 orthologous genes including 94 chemosensory loci from five different gene families (Odorant Receptors, Ionotropic Receptors, Gustatory Receptors, Odorant Binding Proteins, and Chemosensory Proteins). Our results revealed that orthologs with signatures of divergent selection between E. dilemma and E. viridissima were significantly enriched for chemosensory genes. Notably, elevated signals of divergent selection were almost exclusively observed among chemosensory receptors (i.e. Odorant Receptors). Conclusions Our results suggest that rapid changes in the chemosensory gene family occurred among closely related species of orchid bees. These findings are consistent with the hypothesis that strong divergent selection acting on chemosensory receptor genes plays an important role in the evolution and diversification of insect pheromone systems.

Foraging bumblebees leave chemical substances when visiting flowers and the detection of these “scent marks” improves their foraging efficiency. Whereas laboratory studies found that scent-marks convey attraction to food sources, all field studies found foragers to be repelled by recently visited flowers. In this study we aim to resolve this conflict by implementing near-natural reward dynamics in a laboratory feeder array. When feeders were filled with small, non-replenished amounts of reward, worker bumblebees (Bombus terrestris) avoided revisiting the depleted feeders. As evidenced by a “corolla” replacement experiment, feeder discrimination was based on the perception of chemical cues deposited during previous visits. Pentane extracts of bumblebee tarsi acted as a repellent when applied to glass corollas, whereas pure pentane did not. We suggest that scent-marks are simple cues inherent to footprints and emphasize the importance of context to how these cues are interpreted by foraging bees.

Male orchid bees (Euglossini) collect fragrances from flowers and other natural sources, a behaviour that has shaped the euglossine pollination syndrome. Males store such chemicals in hind leg pouches and later expose them during courtship display. In the present study, we show that complex bouquets of two sympatric species of Eulaema, E. meriana and E. bombiformis, are chemically distinct. When exposed during bioassays at display perches individual hind leg extracts rapidly and consistently attracted other males of the correct species, even if derived from males of disparate localities (French Guiana and Panama). Conspecific males as well as females of E. bombiformis arrived at natural perch sites only from downwind, and two copulations were observed. Our findings demonstrate that acquired odours mediate exclusive attraction within species and support the idea that such fragrances are pheromone analogues. Their role in acquiring matings and during male-male interaction is discussed.

Many insects are known to leave lipid footprints while walking on smooth surfaces. Presumably, the deposited substances improve tarsal adhesion. In bumblebees, footprint hydrocarbons also function as scent marks that allow detection and avoidance of recently depleted flowers. I used GC-MS to detect hydrocarbons deposited by bumblebee (Bombus pascuorum) on flowers of Lamium maculatum. In addition to the plants' own cuticular lipids, extracts of corollas that had been visited by bumblebees contained odd-numbered alkenes. The amount of pentacosenes (C25H50) on corollas was linearly related to the number of bumblebee visits, with workers depositing approximately 16 ng per visit (extrapolated to a total of 65 ng of bumblebee cuticular hydrocarbons). Pentacosenes were retained on visited flowers without loss for 2 hr, and probably longer. This and results from flight cage experiments suggest that flower epicuticles retain a chemical record of pollinator visitation, including information on visiting bee species. Continuous footprint accumulation necessitates new explanations concerning the reversibility of "repellent scent marks" of bumblebees.

Male neotropical orchid bees (Euglossini) collect odoriferous substances from orchids and other sources and store them in tibial pouches, accumulating complex and species-specific bouquets. These fragrances are later exposed at display sites, presumably to attract females or conspecific males or both. We hypothesized that the necessity to detect and recognize specific fragrance bouquets has led to peripheral chemosensory specializations in different species of orchid bees. To test this, excised male antennae of four species of Euglossa were stimulated with complete tibial extracts of the same four species in a crosswise experiment. In the majority of the tested extracts, the amplitude of the electroantennogram (EAG) response was significantly different between species and always maximal in males of the extracted species. This effect did not appear to result from a given species' increased sensitivity toward certain attractive components: gas chromatography with electroantennographic detection (GC-EAD) of one extract of Euglossa tridentata evoked similar and generalized response patterns in all four species, encompassing a total of 34 peaks that elicited antennal responses. Therefore, the species effect in EAG responses to complete extracts likely resulted from species-specific interactions of compounds at the receptor level. Antennal specialization to conspecific bouquets adds additional strength to the argument that specificity is an important evolutionary aspect of euglossine tibial fragrances.

Male orchid bees (Euglossini) are attracted to floral and non-floral odours, which they collect and accumulate in hind tibial cavities for subsequent exposure during courtship. Fragrance preferences are species-specific, leading to relatively specialised pollination of euglossophilous plants. We tested the hypothesis that preferences for attractive compounds have led to species-specific sensory adaptations that are measurable by electroantennography (EAG). All of 16 synthetic fragrance compounds elicited significant responses on male bee antennae, with some difference of response spectra between individuals of Euglossa spp. and bumblebee (Bombus terrestris) controls, but no difference between three different species of Euglossa. There was no correspondence between a compounds’ attractiveness in baiting assays and the size of its electrophysiological response. Our results strengthen the view that fragrance preferences are largely mediated by processes in higher nervous centres. Peripheral sensory tuning to single attractive odorants may be constrained by the need to detect and discriminate between many fragrances, including many that have repellent effects on male bees.

The abundance and species richness of highly social bees (Apinae: Meliponini, Apini) were studied at 12 dipterocarp forest sites in Sabah, northern Borneo, using standardized honey-water baiting assays. The 12 sites were grouped in five localities varying in the degree and history of disturbance from selective logging. A baiting assay consisted of spraying diluted honey on vegetation along transects and recording bee arrival on the same day. Repeated assays of the same sites were run in two consecutive years, 1998 and 1999. One-way repeated-measures ANOVAs were calculated on bee abundance and species richness, with locality and year as independent factors. There were significant or marginally significant effects of forest locality on all measures of bee abundance and species richness. However, there were also significant interactions between year and locality on these variables, thwarting conclusions on the effects of logging on bee communities. The diverging results of the two years are likely related to spatio-temporal variation of flowering activity prior to and during the assays, with between-year differences being amplified by pronounced differences in climatic conditions (ENSO). Local or regional differences in flowering phenology complicate the application of resource-based baiting techniques for between-site comparisons of bee diversity.

Nesting habits of highly social stingless bees (Meliponini) were studied in lowland dipterocarp forests in Sabah, Borneo. A total of 275 nests of 12 species of bees were located. All nests were closely associated with living (91.5%) or dead (8.5%) trees, either within pre-formed cavities in the trunk (cavity nests) or situated in or under the tree base (base nests). Species of bees differed in nesting habit. The majority of species (seven) were cavity nesters, but the majority of nests (81%) were base nests. Nests were often aggregated (mean of 1.94 nests/nest tree), with up to eight colonies and three species in a single tree. Nest trees were mostly large to very large (86.1% above 60 cm dbh) commercial timber trees; 47.3% of nest trees were dipterocarps. According to visual inspection nest trees were of significantly lower expected timber quality then randomly chosen control trees. Taking into account information on tree species, size and expected timber quality, we estimated that 34.0 or 42.6% of nest trees were potential harvest trees, depending on harvesting regulations (reduced impact logging (RIL) versus conventional). Lower percentages under RIL guidelines were mostly due to size restrictions that protect very large trees (>120 cm dbh). Harvesting is likely to kill bee colonies associated with the respective tree. Therefore, and because meliponine colonies are long-lived and have low fecundity, direct impact from logging may have lasting effects on bee populations. Harvesting guidelines that retain high proportions of large and hollow trees should be promoted in order to preserve meliponine pollination in sustainable forest management.

Primary tropical lowland rainforest in Sabah, Malaysia, has been largely reduced to small to medium-sized, often isolated, forest islands surrounded by a highly altered agricultural landscape. The biodiversity patterns of leaf litter ant communities were monitored in two forest fragments of differing size as well as in a contiguous forest over the course of two years. Species number and diversity in the forest isolates was significantly lower, reaching only 47.5% of the species number collected in the contiguous forest. Species density was also lower, which had led to a thinning of the ant community in the fragments. Community composition was substantially altered in the forest remnants, and an increase of tramp species with smaller fragment size was detected. These results were unexpected and alarming, as the medium-sized forest is with its 42.9 km2 a comparatively large primary forest fragment for Sabah.

International audience; AbstractMale Neotropical orchid bees collect volatile chemicals from various sources in the environment in order to compose their characteristic perfume bouquets. Amongst other plants, over 600 species of orchids are exclusively pollinated by the bees during their quest for volatiles. Since the plants usually have a scattered distribution, it is assumed that orchid bees can transport the pollinaria across several kilometres due to their flight capabilities and a high dispersal potential. Until now, very long-range flight distances (up to 45 km) of male orchid bees have only been indirectly inferred from habitat requirements of orchids whose pollinaria were carried by captured males, whereas the distances established by direct measures (mark and recapture, radio telemetry) only span distances of up to around 6 km. The discrepancy between inferred and proven distances led us to readdress the question of dispersal ability of male orchid bees. In this study, we used tag, scratch and chemical marking of large numbers of bees to clarify two aspects: (1) the effect of moderate natural obstacles on dispersal and (2) the possibility of very long-range movements across a terrain lacking such obstacles. Our results suggest that a moderate natural obstacle (a valley separating opposite ridges) does not restrain orchid bee movements. Individual bees achieved extraordinary flight distances of more than 50 km across even terrain, extending the directly proven flight distances of male bees by more than an order of magnitude.

Bees make use of plant substrates in more ways than any other group of insects, which is probably linked to their diversification and ecological success. The highly diverse Megachilidae use a wide range of plant-derived brood cell-building materials, including plant leaves, mortar made from plant tissue, resin, plant hairs (“plant wool”), and plant trichome secretions. While certain plant-derived materials are believed to protect nests against microbial decay, this has not been tested, and the factors driving diversification of nesting materials are poorly understood. Here, we investigated the protective effects of plant-derived extrafloral trichome secretions which female European wool-carder bees, Anthidium manicatum, smear on their brood cells. By breeding bees in cages with differential resource supply we generated brood cells with or without trichome secretions. Brood cells with trichome secretions were less attractive to chalcidoid wasps (Monodontomerus obscurus), potential parasites of A. manicatum, in Y-maze olfactometer tests. Also, when exposed at sites where A. manicatum occurs in natural populations, brood cells smeared with trichome secretions were less affected by wasp (Melittobia acasta) parasitism than those without trichome secretions. On the other hand, trichome secretions did not prevent the growth of mold on brood cells, and larval mortality due to microbial decay was almost non-existent even when brood cells were exposed to rainfall. We conclude that the use of plant trichome secretion for brood cell construction has more likely evolved to create enemy-free space in response to chalcidoid wasp parasitism. Parasitoids are likely an underestimated cause of the diversification of cell construction materials and adaptive radiation in megachilid bees.

We measured the nest density of stingless bees (Apidae, Meliponini) in undisturbed and logged-over dipterocarp forests in Sabah, northern Borneo, and eval- uated hypotheses on proximate factors leading to the ob- served variation: population control mediated by (1) nest predation, (2) limitation of nest trees, or (3) food limita- tion. Per-area nest density varied twentyfold across 14 forest sites and was significantly affected by locality, but not by the degree and history of disturbance. Nest densi- ty was generally high in sites located in the Sepilok For- est fragment (mean 8.4 nests/ha), bordering mangroves or plantations. In contrast, nest densities in continu- ous forests were all low (between 0 and 2.1 nests/ha, mean 0.5 nests/ha). Yearly nest mortality was low (13.5-15.0%) over 4 years of observation and did not vary between forest localities, thus limiting the potential of nest predation (1) in creating the observed variation in nest density. The presence of potential nest trees (2), though positively correlated with nest density, explained only a minute fraction of the observed variation. Nest density was best explained by differences in the pollen resources (3) available to the bees (quantified by analy- sis of pollen in bee garbage). Across five selected sites the amount of nonforest pollen (from mangrove or crop plants) included in diets of Trigona collina was positive- ly correlated with T. collina nest density. External pollen sources are a likely supplement to bee diets at times when little flowering occurs inside the forest, thus in- creasing overall bee carrying capacity. Pollen limitation was also indicated by direct measurements of pollen im- port and foraging activity of T. collina in three selected sites: Pollen traps installed at nests in high-density Sep- ilok captured significantly more corbicular pollen than colonies in low-density Deramakot. At the same time, morning foraging activity was also greater in Sepilok, in- dicating a regulatory increase in foraging in response to high pollen availability. We conclude that the abundance of stingless bees in forests in Sabah is chiefly dependent on the local availability of food resources. Bee popula- tions strongly benefit from edge effects and increased foraging habitat diversity. Electronic supplementary ma- terial to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00442- 001-0848-6

Individually marked males of two species of Euglossa were sighted repeatedly and over considerable periods of time (up to 44 days) at artificial fragrance baits exposed on Barro Colorado Island (BCI), Panama. Individuals switched between different bait chemicals that are attractive for the respective species, and no bait preferences or individual bait constancy was observed. GC-MS analyses of 153 males of three species showed that individual hind tibiae contain highly variable quantities of a complex and species-specific blend of fragrance compounds, mainly terpenoids and aromatics. In all three species, frequency distributions of individual quantities were strongly skewed towards individuals with small amounts, and individual amount and complexity were positively correlated. Tibial contents of male Euglossa imperialis that were kept alive in a flight cage for 0, 5, 10, or 15 days showed no qualitative or quantitative change over time, suggesting that the fragrances are very efficiently stored in the hind legs. In Euglossa cognata wing wear, an established age correlate of the species, was positively correlated with individual fragrance quantity. Our results suggest that male euglossines forage continuously for a variety of volatiles, store them, and finally acquire large quantities of a complex and specific fragrance bouquet. Both qualitative and quantitative aspects of individual contents are likely to contain information on male phenotypic and genotypic quality.

International audience; Cuticular lipid profiles have been shown to be species specific within many insect genera, allowing a chemotaxonomic classification of individuals. In this study, we analysed the cuticular lipids of the orchid bees Euglossa viridissima Friese and Euglossa dilemma Bembé & Eltz, cryptic sibling species whose taxonomic status has only recently been clarified. Male individuals of both species were obtained from two locations on the Yucatán Peninsula, Mexico, both in the dry and wet seasons. Their hydrocarbon profiles proved to be species specific, irrespective of location and season. They also allowed a correct assignment of a rare morph of E. viridissima which had long obscured the distinction of the two species. Our results suggest that cuticular hydrocarbon profiles may be suited for chemical taxonomy of orchid bees and might provide an additional clue for difficult species distinctions.

The relative effects of foraging benefits (food intake) and costs (disturbance = pit destruction) on growth rate and foraging behavior of larvae of the ant-lionMyrmeleon mobilis were investigated. In a laboratory experiment second-instar larvae were subjected to combinations of three rates of feeding and three rates of disturbance. Pit relocations were generally rare but occurred more often in starved larvae. Disturbance rate had no significant effect on pit relocation rate. Feeding rate was a major determinant of the energy allocated in pit construction and maintenance (positive relationship). In a second experiment third-instar larvae encountered changing rates of disturbance. Pit size was reduced as a response to increased rates of disturbance. Disturbance had no detectable effect on growth. In general,M. mobilis larvae were cost-conservative, foragers.

Male orchid bees (Euglossini, Apidae, Hymenoptera) expose species-specific blends of volatile chemicals (perfume bouquets) during their courtship display. The perfumes are acquired by collecting fragrant substances from environmental sources, which are then accumulated in specialised hind leg pouches. To balance the perfume composition, the males need to find and collect the required substances in specific relative amounts while facing seasonal, local or habitat-dependent differences in compound availability. Experience-dependent choice of odours, i.e. ‘learned avoidance’ of recently collected components, has been proposed as the mechanism that mediates the accumulation of the stereotypical compound ratios. In the present study, we used the presence of certain compounds in male hind leg pouches as proxy for the respective local compound availability, and investigated whether differences in content are correlated with differences in chemical choice assays. Our results suggest that volatile availability differs between localities (n = 16) as well as habitats (n = 2; coastal vs. inland) across the Yucatan peninsula, Mexico, for both studied species. Male Euglossa dilemma showed a pronounced preference for benzyl benzoate and eugenol at locations where those compounds were rare in hind leg extracts, as predicted by the learned avoidance model. No equivalent correlations were found for Euglossa viridissima. This is the first study to combine chemical analyses of perfumes with bioassays of odour choice. It strengthens the view that negative feedback from collected odours modifies future chemical choice and helps males to acquire specific perfume blends.

In the course of a study on pollen diets of three sympatric species of stingless bees (Apidae: Meliponini) in Sabah, Malaysia, we made the observation that large fractions of the foragers of three colonies of Trigona collina collected corbicular loads of fungal spores in lieu of pollen. Collection of spores continued for at least three consecutive days. The spores were brought to germination in the laboratory and the culture was identified as mold of the genus Rhizopus (Zygomycota, Mucorales). Our observations represent the first reported case of the collection of Rhizopus mold spores in lieu of pollen by bees and a rare case of the collection of fungal spores by bees other than honeybees (Apis).

Most flowering plants establish mutualistic associations with insect pollinators to facilitate sexual reproduction. However, the evolutionary processes that gave rise to these associations remain poorly understood. We reconstructed the times of divergence, diversification patterns, and interaction networks of a diverse group of specialized orchids and their bee pollinators. In contrast to a scenario of coevolution by race formation, we show that fragrance-producing orchids originated at least three times independently after their fragrance-collecting bee mutualists. Whereas orchid diversification has apparently tracked the diversification of orchids’ bee pollinators, bees appear to have depended on the diverse chemical environment of neotropical forests. We corroborated this apparent asymmetrical dependency by simulating co-extinction cascades in real interaction networks that lacked reciprocal specialization. These results suggest that the diversification of insect-pollinated angiosperms may have been facilitated by the exploitation of preexisting sensory biases of insect pollinators.