Land-use-associated stressors interact to reduce bumblebee health at the individual and colony level.

In agricultural landscapes, bees face a variety of stressors, including insecticides and poor-quality food. Although both stressors individually have been shown to affect bumblebee health negatively, few studies have focused on stressor interactions, a scenario expected in intensively used agricultural landscapes. Using the bumblebee Bombus terrestris, a key pollinator in agricultural landscapes, we conducted a fully factorial laboratory experiment starting at nest initiation. We assessed the effects of food quality and insecticides, alone and in interaction, on health traits at various levels, some of which have been rarely studied. Pollen with a diluted nutrient content (low quality) reduced ovary size and delayed colony development. Wing asymmetry, indicating developmental stress, was increased during insecticide exposure and interactions with poor food, whereas both stressors reduced body size. Both stressors and their interaction changed the workers' chemical profile and reduced worker interactions and the immune response. Our findings suggest that insecticides combined with nutritional stress reduce bumblebee health at the individual and colony levels, thus possibly affecting colony performance, such as development and reproduction, and the stability of plant-pollinator networks. The synergistic effects highlight the need of combining stressors in risk assessments and when studying the complex effects of anthropogenic stressors on health outcomes.

Neural and behavioural responses of the pollen-specialist bee Andrena vaga to Salix odours.

An effective means of finding food is crucial for organisms. Whereas specialized animals select a small number of potentially available food sources, generalists use a broader range. Specialist (oligolectic) bees forage on a small range of flowering plants for pollen and use primarily olfactory and visual cues to locate their host flowers. So far, however, little is known about the specific cues oligoleges use to discriminate between hosts and non-hosts and how floral scent compounds of hosts and non-hosts are processed in the bees' olfactory system. In this study, we recorded physiological responses of the antennae (electroantennographic detection coupled to gas chromatography; GC-EAD) and in the brain (optical imaging; GC imaging), and studied host-finding behaviour of oligolectic Andrena vaga bees, a specialist on Salix plants. In total, we detected 37 physiologically active compounds in host and non-host scents. 4-Oxoisophorone, a common constituent in the scent of many Salix species, evoked strong responses in the antennal lobe glomeruli of A. vaga, but not the generalist honeybee Apis mellifera. The specific glomerular responses to 4-oxoisophorone in natural Salix scents reveals a high degree of specialization in A. vaga for this typical Salix odorant component. In behavioural experiments, we found olfactory cues to be the key attractants for A. vaga to Salix hosts, which are also used to discriminate between hosts and non-hosts, and A. vaga demonstrated a behavioural activity for 4-oxoisophorone. A high sensitivity to floral scents enables the specialized bees to effectively find flowers and it appears that A. vaga bees are highly tuned to 4-oxoisophorone at a very low concentration.

Floral traits are associated with the quality but not quantity of heterospecific stigmatic pollen loads.

BACKGROUND: In flowering communities, plant species commonly share pollinators and therefore plant individuals receive heterospecific pollen (HP). However, the patterns of HP transfers can deviate from patterns of plant-pollinator visitations. Although flower-visitor interactions are known to be mediated by floral traits, e.g. floral size or nectar tube depth, the explanatory power of these traits for HP transfer patterns remains elusive. Here, we have explored pollen transfer patterns at three sites in Southern Germany on three dates (early, mid and late summer). At the plant level, we tested whether flower abundance and floral traits are correlated with HP reception and donation. At the community level, we determined whether flower and bee diversity are correlated with network modularity and whether floral traits explain the module affiliation of plant species. We collected the stigmas of flowering plant species, analysed HP and conspecific pollen (CP) loads and measured floral traits, flower and bee diversity. RESULTS: Our results show that the degree and intensity of HP reception or donation at the plant level do not correlate with floral traits, whereas at the community level, the module affiliation of who is sharing pollen with whom is well-explained by floral traits. Additionally, variation in network modularity between communities is better explained by plant diversity and abundance than by bee diversity and abundance. CONCLUSIONS: Overall, our results indicate that floral traits that are known to mediate flower-visitor interactions can improve our understanding of qualitative HP transfer but only provide limited information about the quantity of HP transfer, which more probably depends on other floral traits, flower-visitor identity or community properties.

Bumblebee Behavior on Flowers, but Not Initial Attraction, Is Altered by Short-Term Drought Stress.

Climate change is leading to increasing drought and higher temperatures, both of which reduce soil water levels and consequently water availability for plants. This reduction often induces physiological stress in plants, which in turn can affect floral development and production inducing phenotypic alterations in flowers. Because flower visitors notice and respond to small differences in floral phenotypes, changes in trait expression can alter trait-mediated flower visitor behavior. Temperature is also known to affect floral scent emission and foraging behavior and, therefore, might modulate trait-mediated flower visitor behavior. However, the link between changes in flower visitor behavior and floral traits in the context of increasing drought and temperature is still not fully understood. In a wind-tunnel experiment, we tested the behavior of 66 Bombus terrestris individuals in response to watered and drought-stressed Sinapis arvensis plants and determined whether these responses were modulated by air temperature. Further, we explored whether floral traits and drought treatment were correlated with bumblebee behavior. The initial attractiveness of drought and watered plants did not differ, as the time to first visit was similar. However, bumblebees visited watered plants more often, their visitation rate to flowers was higher on watered plants, and bumblebees stayed for longer, indicating that watered plants were more attractive for foraging. Bumblebee behavior differed between floral trait expressions, mostly independently of treatment, with larger inflorescences and flowers leading to a decrease in the time until the first flower visit and an increase in the number of visits and the flower visitation rate. Temperature modulated bumblebee activity, which was highest at 25°C; the interaction of drought/water treatment and temperature led to higher visitation rate on watered plants at 20°C, possibly as a result of higher nectar production. Thus, bumblebee behavior is influenced by the watered status of plants, and bumblebees can recognize differences in intraspecific phenotypes involving morphological traits and scent emission, despite overall morphological traits and scent emission not being clearly separated between treatments. Our results indicate that plants are able to buffer floral trait expressions against short-term drought events, potentially to maintain pollinator attraction.

Natural Compounds as Spider Repellents: Fact or Myth?

Although some spiders are globally invasive, found at high densities, and may be considered pests (particularly those that are toxic to humans), there are few pest management methods based on experimental data. 'Common wisdom' and advertisements on internet websites assert that a number of natural substances repel spiders. We tested whether the three substances cited most frequently (lemon oil, peppermint oil, and chestnut-fruits) effectively repelled female spiders or whether these were myths. We presented each of the putative repellents versus a control in a two-choice assay and tested responses of females of three invasive spider species in two different families: theridiids, Latrodectus geometricus C. L. Koch (Araneae: Theridiidae) and Steatoda grossa C. L. Koch (Araneae: Theridiidae) and the araneid, Araneus diadematus Clerck . Chestnuts (Araneae: Araneidae) and mint oil strongly repelled L. geometricus and A. diadematus. S. grossa was less sensitive to these chemicals but had a slight tendency to avoid chestnuts. However, lemon oil, the substance most likely to be cited as a repellent (over 1,000,000 hits on Google), had no effect on any of these spiders. We conclude that volatiles released by mint oil and chestnuts may be effective in deterring spider settlement in two different families of spiders, but lemon oil as a repellent is a myth.

Sexual Deception in the Eucera-Pollinated Ophrys leochroma: A Chemical Intermediate between Wasp- and Andrena-Pollinated Species.

Ophrys flowers mimic sex pheromones of attractive females of their pollinators and attract males, which attempt to copulate with the flower and thereby pollinate it. Virgin females and orchid flowers are known to use the same chemical compounds in order to attract males. The composition of the sex pheromone and its floral analogue, however, vary between pollinator genera. Wasp-pollinated Ophrys species attract their pollinators by using polar hydroxy acids, whereas Andrena-pollinated species use a mixture of non-polar hydrocarbons. The phylogeny of Ophrys shows that its evolution was marked by episodes of rapid diversification coinciding with shifts to different pollinator groups: from wasps to Eucera and consequently to Andrena and other bees. To gain further insights, we studied pollinator attraction in O. leochroma in the context of intra- and inter-generic pollinator shifts, radiation, and diversification in the genus Ophrys. Our model species, O. leochroma, is pollinated by Eucera kullenbergi males and lies in the phylogeny between the wasp and Andrena-pollinated species; therefore, it is a remarkable point to understand pollinator shifts. We collected surface extracts of attractive E. kullenbergi females and labellum extracts of O. leochroma and analyzed them by using gas chromatography with electroantennographic detection (GC-EAD) and gas chromatography coupled with mass spectrometry (GC-MS). We also performed field bioassays. Our results show that O. leochroma mimics the sex pheromone of its pollinator's female by using aldehydes, alcohols, fatty acids, and non-polar compounds (hydrocarbons). Therefore, in terms of the chemistry of pollinator attraction, Eucera-pollinated Ophrys species might represent an intermediate stage between wasp- and Andrena-pollinated orchid species.

Telipogon peruvianus (Orchidaceae) Flowers Elicit Pre-Mating Behaviour in Eudejeania (Tachinidae) Males for Pollination.

Several neotropical orchid genera have been proposed as being sexually deceptive; however, this has been carefully tested in only a few cases. The genus Telipogon has long been assumed to be pollinated by male tachinid flies during pseudocopulatory events but no detailed confirmatory reports are available. Here, we have used an array of methods to elucidate the pollination mechanism in Telipogon peruvianus. The species presents flowers that have a mean floral longevity of 33 days and that are self-compatible, although spontaneous self-pollination does not occur. The flowers attract males of four tachinid species but only the males of an undescribed Eudejeania (Eudejeania aff. browni; Tachinidae) species are specific pollinators. Males visit the flowers during the first few hours of the day and the pollination success is very high (42% in one patch) compared with other sexually deceptive species. Female-seeking males are attracted to the flowers but do not attempt copulation with the flowers, as is usually described in sexually deceptive species. Nevertheless, morphological analysis and behavioural tests have shown an imperfect mimicry between flowers and females suggesting that the attractant stimulus is not based only on visual cues, as long thought. Challenging previous conclusions, our chemical analysis has confirmed that flowers of Telipogon release volatile compounds; however, the role of these volatiles in pollinator behaviour remains to be established. Pollinator behaviour and histological analyses indicate that Telipogon flowers possess scent-producing structures throughout the corolla. Our study provides the first confirmed case of (i) a sexually deceptive species in the Onciidinae, (ii) pollination by pre-copulatory behaviour and (iii) pollination by sexual deception involving tachinid flies.

Orchid mimics honey bee alarm pheromone in order to attract hornets for pollination.

Approximately one-third of the world's estimated 30,000 orchid species are deceptive and do not reward their pollinators with nectar or pollen. Most of these deceptive orchids imitate the scent of rewarding flowers or potential mates. In this study, we investigated the floral scent involved in pollinator attraction to the rewardless orchid Dendrobium sinense, a species endemic to the Chinese island Hainan that is pollinated by the hornet Vespa bicolor. Via chemical analyses and electrophysiological methods, we demonstrate that the flowers of D. sinense produce (Z)-11-eicosen-1-ol and that the pollinator can smell this compound. This is a major compound in the alarm pheromones of both Asian (Apis cerana) and European (Apis mellifera) honey bees and is also exploited by the European beewolf (Philanthus triangulum) to locate its prey. This is the first time that (Z)-11-eicosen-1-ol has been identified as a floral volatile. In behavioral experiments, we demonstrate that the floral scent of D. sinense and synthetic (Z)-11-eicosen-1-ol are both attractive to hornets. Because hornets frequently capture honey bees to feed to their larvae, we suggest that the flowers of D. sinense mimic the alarm pheromone of honey bees in order to attract prey-hunting hornets for pollination.

Ménage à trois-two endemic species of deceptive orchids and one pollinator species.

In the sexually deceptive orchid genus Ophrys, reproductive isolation is based on the specific attraction of males of a single pollinator species by mimicking the female species-specific sex pheromone. Changes in the odor composition can lead to hybridization and speciation by the attraction of a new pollinator that acts as an isolation barrier toward other sympatrically occurring Ophrys species. On Sardinia, we investigated the evolutionary origin of two sympatrically occurring endemic species, Ophrys chestermanii and O. normanii, which are both pollinated by males of the cuckoo bumblebee Bombus vestalis. Chemical and electrophysiological analyses of floral scent and genetic analyses with amplified fragment length polymorphisms and plastid-markers clearly showed that O. normanii is neither a hybrid nor a hybrid species. The two species evolved from different ancestors, viz. O. normanii from O. tenthredinifera and O. chestermanii from O. annae, and converged to the same pollinator attracted by the same bouquet of polar compounds. In spite of sympatry, pollinator sharing and overlapping blooming periods, no evidence has been obtained for gene flow between O. chestermanii and O. normanii indicating an unusual case among sexually deceptive orchids in which postmating rather than premating reproductive isolation mechanisms strongly prevent interspecific gene flow.