Tropical and montane Apis cerana show distinct dance-distance calibration curves.

Social bees have evolved sophisticated communication systems to recruit nestmates to newly found food sources. As foraging ranges can vary from a few hundred meters to several kilometers depending on the environment or season, populations of social bee species living in different climate zones likely show specific adaptations in their recruitment communication. Accordingly, studies in the western honey bee, Apis mellifera, demonstrated that temperate populations exhibit shallower dance-calibration curves compared to tropical populations. Here we report the first comparison of calibration curves for three Indian A. cerana lineages: the tropical A. indica, and the two montane Himalayan populations A. c. cerana (Himachal Pradesh) and A. c. kashmirensis (Jammu and Kashmir). We found that the colonies of the two montane A. cerana populations show dance-distance calibration curves with significantly shallower slopes than the tropical A. indica. Next, we transferred A. c. cerana colonies to Bangalore (∼ 2600 km away) to obtain calibration curves in the same location as A. indica. The common garden experiment confirmed this difference in slopes, implying that the lineages exhibit genetically fixed differences in dance-distance coding. However, the slopes of the calibration curves of the transferred A. c. cerana colonies were also significantly higher than those tested in Himachal Pradesh indicating an important effect of the environment. The differences in dance-distance coding between temperate and tropical A. cerana lineages resemble those described for A. mellifera suggesting that populatiInons of both species independently evolved similar adaptations.

Cuticular Hydrocarbon Profiles of Himalayan Bumble Bees (Hymenoptera: Bombus Latreille) are Species-Specific and Show Elevational Variation.

Bumble bees are important pollinators in natural environments and agricultural farmlands, and they are in particular adapted to harsh environments like high mountain habitats. In these environments, animals are exposed to low temperature and face the risk of desiccation. The Eastern Himalayas are one of the recognized biodiversity hotspots worldwide. The area covers subtropical rainforest with warm temperature and high precipitation as well as high mountain ranges with peaks reaching up to 7,000 m, shaping a diverse floral and faunal community at the different elevational zones. To identify possible adaptation strategies, we investigated the cuticular hydrocarbon profiles of four bumble bee species occurring at different elevational ranges in Arunachal Pradesh, the northeastern most state in India. At 17 locations along an elevational gradient, we collected workers of two species from lower elevations (B. albopleuralis and B. breviceps; ~ 100 m - 3,000 m asl) and two species from higher elevations (B. prshewalskyi and B. mirus; ~ 2,800 m - 4,500 m asl). The CHC profiles of all four species showed a significant degree of variation in the composition of hydrocarbons, indicating species specificity. We also found clear correlation with elevation. The weighted mean chain length of the hydrocarbons significantly differed between the low and high elevation species, and the proportion of saturated hydrocarbons in CHC profiles significantly increased with the elevational range of the bumble bee species. Our results indicate that bumble bees living at high elevations reduce the risk of water loss by adapting their CHC composition on their cuticle, a phenomenon that has also been found in other insects like ants and fruit flies.

Tracking Sugar-Elicited Local Searching Behavior in Drosophila.

Foraging behavior is essential for the survival of organisms as it enables them to locate and acquire essential food resources. In Drosophila, hunger triggers a distinct search behavior following the consumption of small quantities of a sugar solution. This report presents a simple experimental setup to study sugar-elicited search behavior with the aim of uncovering the underlying mechanisms. Minute quantities of concentrated sugar solution elicit sustained searching behavior in flies. The involvement of path integration in this behavior has been established, as flies utilize their trajectory to return to the sugar location. The most recent findings provide evidence of temporal modulation in the initiation and intensity of the search behavior after sugar intake. We have also used this setup for artificial activation of specific taste-receptor neurons in the pharynx, which elicits the search behavior. The Drosophila neurogenetic toolkit offers a diverse array of tools and techniques that can be combined with the sugar-elicited search behavior paradigm to study the neural and genetic mechanisms underlying foraging. Understanding the neural basis of hunger-driven searching behavior in flies contributes to the field of neurobiology as a whole, offering insights into the regulatory mechanisms that govern feeding behaviors not only in other organisms but also in humans.

Nutritional properties of giant water bug, Lethocerus indicus a traditional edible insect species of North-East India.

Edible insects play an important role in human health and food security. Among those, the Giant water bug, Lethocerus indicus (Lep.& Ser.) is a widely used edible insect known for its aroma, flavor, and therapeutic purposes. In the present study, we investigated the nutritional profile, natural habitat, and feeding behavior of L. indicus in aquarium conditions. A comparative analysis of male and female insects' aroma contents and fatty acid (FA) profiles was also conducted. A dry fried male insect yielded volatile oil of 0.96%/2 g body weight, whereas a dry fried female yielded 0.48%/5.36 g of body weight. In terms of lipids, fresh male insects had 0.15%/5.42 g of body weight and fresh female insects had 0.28%/9.48 g of body weight. There are 24 volatile compounds specific to males, 37 specific to females, and 13 commons to both were identified. 2-Hexen-1-ol, acetate, (Z)- which smells like banana, was prevalently found in males while 4-Octene, 2,6-dimethyl-, [S-(Z)] was prevalently found in female insects. Fatty acids profile analysis detected 32 FA with 12 unique FA from males whereas 22 FA and 3 unique FA were identified from female insects. The SFA percentage present in males and females was 77.44% and 85.21%. Males had 6.78% MUFA content while females have 4.75%. Males have 18% PUFA content enriched with DHA, and EPA, while females had 10.04%. This study revealed that with the presence of a banana-like smell of volatile compound and more MUFA and PUFA in males, the native people of North-East India preferred male over female insects for entomophagy.

Temporal effects of sugar intake on fly local search and honey bee dance behaviour.

Honey bees communicate flight navigational information of profitable food to nestmates via their dance, a small-scale walking pattern, inside the nest. Hungry flies and honey bee foragers exhibit a sugar-elicited search involving path integration that bears a resemblance to dance behaviour. This study aimed to investigate the temporal dynamics of the initiation of sugar-elicited search and dance behaviour, using a comparative approach. Passive displacement experiments showed that feeding and the initiation of search could be spatially and temporally dissociated. Sugar intake increased the probability of initiating a search but the actual onset of walking triggers the path integration system to guide the search. When prevented from walking after feeding, flies and bees maintained their motivation for a path integration-based search for a duration of 3 min. In flies, turning and associated characters were significantly reduced during this period but remained higher than in flies without sugar stimulus. These results suggest that sugar elicits two independent behavioural responses: path integration and increased turning, with the initiation and duration of path integration system being temporally restricted. Honey bee dance experiments demonstrated that the motivation of foragers to initiate dance persisted for 15 min, while the number of circuits declined after 3 min following sugar ingestion. Based on these findings, we propose that food intake during foraging increases the probability to initiate locomotor behaviours involving the path integration system in both flies and honey bees, and this ancestral connection might have been co-opted and elaborated during the evolution of dance communication by honey bees.

Diverse communication strategies in bees as a window into adaptations to an unpredictable world.

Communication is a fundamental feature of animal societies and helps their members to solve the challenges they encounter, from exploiting food sources to fighting enemies or finding a new home. Eusocial bees inhabit a wide range of environments and they have evolved a multitude of communication signals that help them exploit resources in their environment efficiently. We highlight recent advances in our understanding of bee communication strategies and discuss how variation in social biology, such as colony size or nesting habits, and ecological conditions are important drivers of variation in communication strategies. Anthropogenic factors, such as habitat conversion, climate change, or the use of agrochemicals, are changing the world bees inhabit, and it is becoming clear that this affects communication both directly and indirectly, for example by affecting food source availability, social interactions among nestmates, and cognitive functions. Whether and how bees adapt their foraging and communication strategies to these changes represents a new frontier in bee behavioral and conservation research.

Increased complexity of worker CHC profiles in Apis dorsata correlates with nesting ecology.

Cuticular hydrocarbons (CHC) are known to serve as discrimination cues and will trigger defence behaviour in a plethora of eusocial insects. However, little is known how about nestmate recognition ability selects for CHC diversification. In this study we investigate differences in CHC composition of four major honey bee species with respect to the differences in their nesting behavior. In contrast to A. mellifera, A. cerana and A. florea, the giant honey bee A. dorsata prefers to build their nests in aggregations with very small spatial distances between nests, which increases the probability of intrusions. Thus, A. dorsata exhibits a particularly challenging nesting behavior which we hypothesize should be accompanied with an improved nestmate recognition system. Comparative analyses of the worker CHC profiles indicate that A. dorsata workers exhibit a unique and more complex CHC profile than the other three honey bee species. This increased complexity is likely based on a developmental process that retains the capability to synthesize methyl-branched hydrocarbons as adults. Furthermore, two sets of behavioral experiments provide evidence that A. dorsata shows an improved nestmate discrimination ability compared to the phylogenetically ancestral A. florea, which is also open-nesting but does not form nest aggregations. The results of our study suggest that ecological traits like nesting in aggregation might be able to drive CHC profile diversification even in closely related insect species.

A hard day's night: Patterns in the diurnal and nocturnal foraging behavior of Apis dorsata across lunar cycles and seasons.

The giant honey bee Apis dorsata is unusual in being able to forage during both the day and the night. To date, the extent of this unique nocturnal foraging behavior and the environmental factors correlating with it have not been deeply investigated. We conducted the first systematic investigation into the nocturnal behavior of A. dorsata in Southern India by tracking the daily and nightly foraging activity of A. dorsata colonies in an urban environment for 8 months, over multiple seasons and lunar cycles. We found strong evidence that A. dorsata can behave in a manner that is "cathemeral" (active over the entire diel cycle) when environmental illumination is sufficient for nocturnal flight. However, workers were not always active even when the environment should have been bright enough for them to forage, suggesting that their nocturnal foraging behavior was also affected by seasonal changes in resource availability. The foraging activity observed during the day versus twilight versus night differed between seasons; notably, nocturnal activity rates were higher than diurnal activity rates during the winter. We found that at our study site A. dorsata routinely exhibits both diurnal and crepuscular activity, foraging just as intensely during the short twilight hours as during the day. The high foraging activity observed during the twilight and nighttime hours shows that A. dorsata colonies can extend their foraging beyond the daylight hours and reveals that foraging during these dimly lit hours is an integral part of their foraging ecology. This evidence of the importance of nocturnal and crepuscular foraging by A. dorsata paves the way for future studies examining the role of this species in nocturnal pollination networks, the contribution of nocturnal foraging to colony-level nutrition and energy budget, and the evolution of this unusual behavior. Future work comparing nocturnal activity in light polluted urban environments versus unpolluted natural environments is particularly encouraged to determine the generalizability of these findings.

Tyramine 1 Receptor Distribution in the Brain of Corbiculate Bees Points to a Conserved Function.

Sucrose represents an important carbohydrate source for most bee species. In the Western honeybee (Apis mellifera) it was shown that individual sucrose responsiveness correlates with the task performed in the colony, supporting the response threshold theory which states that individuals with the lowest threshold for a task-associated stimuli will perform the associated task. Tyramine was shown to modulate sucrose responsiveness, most likely via the tyramine 1 receptor. This receptor is located in brain areas important for the processing of gustatory stimuli. We asked whether the spatial expression pattern of the tyramine 1 receptor is a unique adaptation of honeybees or if its expression represents a conserved trait. Using a specific tyramine receptor 1 antibody, we compared the spatial expression of this receptor in the brain of different corbiculate bee species, including eusocial honeybees, bumblebees, stingless bees, and the solitary bee Osmia bicornis as an outgroup. We found a similar labeling pattern in the mushroom bodies, the central complex, the dorsal lobe, and the gnathal ganglia, indicating a conserved receptor expression. With respect to sucrose responsiveness this result is of special importance. We assume that the tyramine 1 receptor expression in these neuropiles provides the basis for modulation of sucrose responsiveness. Furthermore, the tyramine 1 receptor expression seems to be independent of size, as labeling is similar in bee species that differ greatly in their body size. However, the situation in the optic lobes appears to be different. Here, the lobula of stingless bees is clearly labeled by the tyramine receptor 1 antibody, whereas this labeling is absent in other species. This indicates that the regulation of this receptor is different in the optic lobes, while its function in this neuropile remains unclear.

Distance estimation by Asian honey bees in two visually different landscapes.

Honey bees estimate distances to food sources using image motion experienced on the flight path and they use this measure to tune the waggle phase duration in their dance communication. Most studies on the dance-related odometer are based on experiments with Apis mellifera foragers trained into small tunnels with black and white patterns which allowed quantifiable changes in the optic flow. In this study, we determined the calibration curves of two Asian honey bee species, A. florea and A. cerana, in two different natural environments with clear differences in the vegetation conditions and hence visual contrast. We found that the dense vegetation condition (with higher contrast) elicited a more rapid increase in the waggle phase duration with distance than the sparse vegetation in A. florea but not in A. cerana Our findings suggest that contrast sensitivity of the waggle dance odometer might vary among honey bee species.

Nesting ecology does not explain slow-fast cognitive differences among honeybee species.

Slow-fast behavioral and life history differences have been tied to slow-fast variation in cognition that is part of the general speed-accuracy tradeoff. While there is growing evidence for such cognitive variation and its association with behavior and life history at the intraspecific level, it is unknown if a similar relationship extends to the interspecific level. Since interspecific differences in cognition have been shown to be a function of ecology and life history, such differences should be reflected in multiple traits that comprise the slow-fast cognitive axis. In this study, by measuring multiple cognitive traits in individuals, we tested for differences in the cognitive phenotype among four honeybee species, which differ in their behavior and life history in a manner that is associated with differences in their nesting ecology. Our results indicate that a set of cognitive traits consistently covary within each species, resulting in slow and fast cognitive phenotypes that largely meet the predictions of the speed-accuracy tradeoff. We also find that the four species group into two distinct clusters on a slow-fast cognitive axis, although their positions do not align with the known differences in their life history and nesting ecology. We instead find that cognitive differences among the four species are correlated with their brain size. We discuss the possible implications of these results for the role of ecology on slow-fast cognitive differences and the evolution of cognition.

Search Behavior of Individual Foragers Involves Neurotransmitter Systems Characteristic for Social Scouting.

In honey bees search behavior occurs as social and solitary behavior. In the context of foraging, searching for food sources is performed by behavioral specialized foragers, the scouts. When the scouts have found a new food source, they recruit other foragers (recruits). These recruits never search for a new food source on their own. However, when the food source is experimentally removed, they start searching for that food source. Our study provides a detailed description of this solitary search behavior and the variation of this behavior among individual foragers. Furthermore, mass spectrometric measurement showed that the initiation and performance of this solitary search behavior is associated with changes in glutamate, GABA, histamine, aspartate, and the catecholaminergic system in the optic lobes and central brain area. These findings strikingly correspond with the results of an earlier study that showed that scouts and recruits differ in the expression of glutamate and GABA receptors. Together, the results of both studies provide first clear support for the hypothesis that behavioral specialization in honey bees is based on adjusting modulatory systems involved in solitary behavior to increase the probability or frequency of that behavior.

Downregulation of the tyrosine degradation pathway extends Drosophila lifespan.

Aging is characterized by extensive metabolic reprogramming. To identify metabolic pathways associated with aging, we analyzed age-dependent changes in the metabolomes of long-lived Drosophila melanogaster. Among the metabolites that changed, levels of tyrosine were increased with age in long-lived flies. We demonstrate that the levels of enzymes in the tyrosine degradation pathway increase with age in wild-type flies. Whole-body and neuronal-specific downregulation of enzymes in the tyrosine degradation pathway significantly extends Drosophila lifespan, causes alterations of metabolites associated with increased lifespan, and upregulates the levels of tyrosine-derived neuromediators. Moreover, feeding wild-type flies with tyrosine increased their lifespan. Mechanistically, we show that suppression of ETC complex I drives the upregulation of enzymes in the tyrosine degradation pathway, an effect that can be rescued by tigecycline, an FDA-approved drug that specifically suppresses mitochondrial translation. In addition, tyrosine supplementation partially rescued lifespan of flies with ETC complex I suppression. Altogether, our study highlights the tyrosine degradation pathway as a regulator of longevity.

Geographical distribution of the giant honey bee Apis laboriosa Smith, 1871 (Hymenoptera, Apidae).

Worldwide pollinator declines have dramatically increased our need to survey and monitor pollinator distributions and abundances. The giant honey bee, Apis laboriosa, is one of the important pollinators at higher altitudes of the Himalayas. This species has a restricted distribution along the Himalayas and neighbouring mountain ranges of Asia. Previous assessments of its distribution, published more than 20 years ago, were based on museum specimens. Since then, 244 additional localities have been revealed through field trips by the authors, publications, and websites. We present a revised distribution for A. laboriosa that better defines its range and extends it eastward to the mountains of northern Vietnam, southward along the Arakan Mountains to west-central Myanmar, into the Shillong Hills of Meghalaya, India, and northwestward in Uttarakhand, India. This species is generally found at elevations between 1000-3000 m a.s.l.. In northeastern India A. laboriosa colonies occur during summer at sites as low as 850 m a.s.l. and some lower elevation colonies maintain their nests throughout the winter. Finally, we report three regions in Arunachal Pradesh, India, and nine locations in northern Vietnam, where we observed workers of A. laboriosa and A. dorsata foraging sympatrically; their co-occurrence supports the species status of Apis laboriosa.

A field-based quantitative analysis of sublethal effects of air pollution on pollinators.

While the impact of air pollution on human health is well studied, mechanistic impacts of air pollution on wild systems, including those providing essential ecosystem services, are largely unknown, but directly impact our health and well-being. India is the world's largest fruit producer, second most populous country, and contains 9 of the world's 10 most polluted cities. Here, we sampled Giant Asian honey bees, Apis dorsata, at locations with varying air pollution levels in Bangalore, India. We observed significant correlations between increased respirable suspended particulate matter (RSPM) deposition and changes in bee survival, flower visitation, heart rate, hemocyte levels, and expression of genes related to lipid metabolism, stress, and immunity. Lab-reared Drosophila melanogaster exposed to these same sites also exhibited similar molecular and physiological differences. Our study offers a quantitative analysis on the current impacts of air pollution on insects, and indicates the urgency for more nonhuman studies to accurately assess the effects of pollution on our natural world.

Sex-specific molecular specialization and activity rhythm-dependent gene expression in honey bee antennae.

We performed an RNA-seq-based comparison of gene expression levels in the antennae of honey bee drones and time-trained foragers (workers) collected at different times of the day and different activity states. Interestingly, olfaction-related genes [i.e. odorant receptor (Or) genes, odorant binding protein (Obp) genes, carboxyl esterase (CEst) genes, etc.] showed stable gene expression differences between drone and worker antennae. Drone antennae showed higher expression of 24 Or genes, of which 21 belong to the clade X which comprises the receptor for the major queen pheromone compound 9-ODA. This high number of drone-biased Or genes suggests that more than previously thought play a role in sex-pheromone communication. In addition, we found higher expression levels for many non-olfaction-related genes including nitric oxide synthase (NOS), and the potassium channel Shaw In contrast, workers showed higher expression of 67 Or genes, which belong to different Or clades that are involved in pheromone communication as well as the perception of cuticular hydrocarbons and floral scents. Further, drone antennae showed higher expression of genes involved in energy metabolism, whereas worker antennae showed higher expression of genes involved in neuronal communication, consistent with earlier reports on peripheral olfactory plasticity. Finally, drones that perform mating flight in the afternoon (innate) and foragers that are trained to forage in the afternoon (adapted) showed similar daily changes in the expression of two major clock genes, period and cryptochrome2 Most of the other genes showing changes with time or onset of daily flight activity were specific to drones and foragers.

Adaptive evolution of honeybee dance dialects.

Efficient communication is highly important for the evolutionary success of social animals. Honeybees (genus Apis) are unique in that they communicate the spatial information of resources using a symbolic 'language', the waggle dance. Different honeybee species differ in foraging ecology but it remains unknown whether this shaped variation in the dance. We studied distance dialects-interspecific differences in how waggle duration relates to flight distance-and tested the hypothesis that these evolved to maximize communication precision over the bees' foraging ranges. We performed feeder experiments with Apis cerana, A. florea and A. dorsata in India and found that A. cerana had the steepest dialect, i.e. a rapid increase in waggle duration with increasing feeder distance, A. florea had an intermediate, and A. dorsata had the lowest dialect. By decoding dances for natural food sites, we inferred that the foraging range was smallest in A. cerana, intermediate in A. florea and largest in A. dorsata. The inverse correlation between foraging range and dialect was corroborated when comparing six (sub)species across the geographical range of the genus including previously published data. We conclude that dance dialects constitute adaptations resulting from a trade-off between the spatial range and the spatial accuracy of communication.

Inter-individual variation in honey bee dance intensity correlates with expression of the foraging gene.

Individual behavioural differences in responding to the same stimuli is an integral part of division of labour in eusocial insect colonies. Amongst honey bee nectar foragers, individuals strongly differ in their sucrose responsiveness, which correlates with strong differences in behavioural decisions. In this study, we explored whether the mechanisms underlying the regulation of foraging are linked to inter-individual differences in the waggle dance activity of honey bee foragers. We first quantified the variation in dance activity amongst groups of foragers visiting an artificial feeder filled consecutively with different sucrose concentrations. We then determined, for these foragers, the sucrose responsiveness and the brain expression levels of three genes associated with food search and foraging; the foraging gene Amfor, octopamine receptor gene AmoctαR1 and insulin receptor AmInR-2. As expected, foragers showed large inter-individual differences in their dance activity, irrespective of the reward offered at the feeder. The sucrose responsiveness correlated positively with the intensity of the dance activity at the higher reward condition, with the more responsive foragers having a higher intensity of dancing. Out of the three genes tested, Amfor expression significantly correlated with dance activity, with more active dancers having lower expression levels. Our results show that dance and foraging behaviour in honey bees have similar mechanistic underpinnings and supports the hypothesis that the social communication behaviour of honey bees might have evolved by co-opting behavioural modules involved in food search and foraging in solitary insects.

Species composition and elevational distribution of bumble bees (Hymenoptera, Apidae, Bombus Latreille) in the East Himalaya, Arunachal Pradesh, India.

The East Himalaya is one of the world's most biodiverse ecosystems. However, very little is known about the abundance and distribution of many plant and animal taxa in this region. Bumble bees are a group of cold-adapted and high elevation insects that fulfil an important ecological and economical function as pollinators of wild and agricultural flowering plants and crops. The Himalayan mountain range provides ample suitable habitats for bumble bees. Systematic study of Himalayan bumble bees began a few decades ago and the main focus has centred on the western region, while the eastern part of the mountain range has received little attention and only a few species have been verified. During a three-year survey, more than 700 bumble bee specimens of 21 species were collected in Arunachal Pradesh, the largest of the north-eastern states of India. The material included a range of species that were previously known from a limited number of collected specimens, which highlights the unique character of the East Himalayan ecosystem. Our results are an important first step towards a future assessment of species distribution, threat, and conservation. Clear elevation patterns of species diversity were observed, which raise important questions about the functional adaptations that allow bumble bees to thrive in this particularly moist region in the East Himalaya.