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.

Laboratory Risk Assessment of Three Entomopathogenic Fungi Used for Pest Control toward Social Bee Pollinators.

The use of fungal-based biopesticides to reduce pest damage and protect crop quality is often considered a low-risk control strategy. Nevertheless, risk assessment of mycopesticides is still needed since pests and beneficial insects, such as pollinators, co-exist in the same agroecosystem where mass use of this strategy occurs. In this context, we evaluated the effect of five concentrations of three commercial entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae, and Cordyceps fumosorosea, by direct contact and ingestion, on the tropical stingless bees Scaptotrigona depilis and Tetragonisca angustula, temperate bee species, the honey bee Apis mellifera, and the bumble bee Bombus terrestris, at the individual level. Furthermore, we studied the potential of two infection routes, either by direct contact or ingestion. In general, all three fungi caused considerable mortalities in the four bee species, which differed in their response to the different fungal species. Scaptotrigona depilis and B. terrestris were more susceptible to B. bassiana than the other fungi when exposed topically, and B. terrestris and A. mellifera were more susceptible to M. anisopliae when exposed orally. Interestingly, increased positive concentration responses were not observed for all fungal species and application methods. For example, B. terrestris mortalities were similar at the lowest and highest fungal concentrations for both exposure methods. This study demonstrates that under laboratory conditions, the three fungal species can potentially reduce the survival of social bees at the individual level. However, further colony and field studies are needed to elucidate the susceptibility of these fungi towards social bees to fully assess the ecological risks.

Organization enhances collective vigilance in the hovering guards of Tetragonisca angustula bees.

One benefit of group living is vigilance against predators. Previous studies have investigated the group size effect, where individual vigilance decreases as group size increases without reducing the overall ability of the group to detect predators. However, there has been comparatively little research on whether the positioning of individuals can improve the collective vigilance of the group. We studied the coordination of vigilance and its effect on predator detection in the eusocial bee Tetragonisca angustula. Nests are defended by hovering guards that detect and intercept intruders before they reach the nest entrance, in addition to those that stand upon it. We show that hovering guards are positioned nonrandomly, with a strong tendency for equal numbers on both sides of the entrance. This organization increases the collective vigilance of the guard group, as groups distributed in an even ratio, either side of the entrance, have a greater collective field of view than groups that deviate from an even ratio. Finally, we use a bioassay to show that when guards are on both sides of the entrance, their ability to detect intruders before they reach the entrance increases. Overall, our results provide strong evidence that vigilance is coordinated and that this improves nest defense. Although other group-living animals are often selfish in their individual vigilance behaviors and face competing time constraints such as foraging, the altruistic nature of eusocial insect workers has probably facilitated the evolution of coordinated vigilance, as documented here in T. angustula.

Diploid Male Production Results in Queen Death in the Stingless Bee Scaptotrigona depilis.

As in most Hymenoptera, the eusocial stingless bees (Meliponini) have a complementary sex determination (CSD) system. When a queen makes a "matched mating" with a male that shares a CSD allele with her, half of their diploid offspring are diploid males rather than females. Matched mating imposes a cost, since diploid male production reduces the colony workforce. Hence, adaptations preventing the occurrence or attenuating its effects are likely to arise. Here we provide clear evidence that in the stingless bee Scaptotrigona depilis, the emergence of diploid males induces queen death, and this usually occurs within 10-20 days of the emergence of diploid male offspring from their pupae. Queens that have not made a matched mating die when introduced into a colony in which diploid males are emerging. This shows that the adult diploid males, and not the queen that has made a matched mating herself, are the proximate cause of queen death. Analysis of the cuticular hydrocarbon profiles of adult haploid and diploid males shows six compounds with significant differences. Moreover, the diploid and haploid males only acquire distinct cuticular hydrocarbon profiles 10 days after emergence. Our data shows that the timing of queen death occurs when the cuticular hydrocarbons of haploid and diploid males differ significantly, suggesting that these chemical differences could be used as cues or signals to trigger queen death.

Hygienic behaviour in Brazilian stingless bees.

Social insects have many defence mechanisms against pests and pathogens. One of these is hygienic behaviour, which has been studied in detail in the honey bee, Apis mellifera Hygienic honey bee workers remove dead and diseased larvae and pupae from sealed brood cells, thereby reducing disease transfer within the colony. Stingless bees, Meliponini, also rear broods in sealed cells. We investigated hygienic behaviour in three species of Brazilian stingless bees (Melipona scutellaris, Scaptotrigona depilis, Tetragonisca angustula) in response to freeze-killed brood. All three species had high mean levels of freeze-killed brood removal after 48 h ∼99% in M. scutellaris, 80% in S. depilis and 62% in T. angustula (N=8 colonies per species; three trials per colony). These levels are greater than in unselected honey bee populations, ∼46%. In S. depilis there was also considerable intercolony variation, ranging from 27% to 100% removal after 2 days. Interestingly, in the S. depilis colony with the slowest removal of freeze-killed brood, 15% of the adult bees emerging from their cells had shrivelled wings indicating a disease or disorder, which is as yet unidentified. Although the gross symptoms resembled the effects of deformed wing virus in the honey bee, this virus was not detected in the samples. When brood comb from the diseased colony was introduced to the other S. depilis colonies, there was a significant negative correlation between freeze-killed brood removal and the emergence of deformed worker bees (P=0.001), and a positive correlation with the cleaning out of brood cells (P=0.0008). This shows that the more hygienic colonies were detecting and removing unhealthy brood prior to adult emergence. Our results indicate that hygienic behaviour may play an important role in colony health in stingless bees. The low levels of disease normally seen in stingless bees may be because they have effective mechanisms of disease management, not because they lack diseases.

Beekeeping practices and geographic distance, not land use, drive gene flow across tropical bees.

Across the globe, wild bees are threatened by ongoing natural habitat loss, risking the maintenance of plant biodiversity and agricultural production. Despite the ecological and economic importance of wild bees and the fact that several species are now managed for pollination services worldwide, little is known about how land use and beekeeping practices jointly influence gene flow. Using stingless bees as a model system, containing wild and managed species that are presumed to be particularly susceptible to habitat degradation, here we examine the main drivers of tropical bee gene flow. We employ a novel landscape genetic approach to analyse data from 135 populations of 17 stingless bee species distributed across diverse tropical biomes within the Americas. Our work has important methodological implications, as we illustrate how a maximum-likelihood approach can be applied in a meta-analysis framework to account for multiple factors, and weight estimates by sample size. In contrast to previously held beliefs, gene flow was not related to body size or deforestation, and isolation by geographic distance (IBD) was significantly affected by management, with managed species exhibiting a weaker IBD than wild ones. Our study thus reveals the critical importance of beekeeping practices in shaping the patterns of genetic differentiation across bee species. Additionally, our results show that many stingless bee species maintain high gene flow across heterogeneous landscapes. We suggest that future efforts to preserve wild tropical bees should focus on regulating beekeeping practices to maintain natural gene flow and enhancing pollinator-friendly habitats, prioritizing species showing a limited dispersal ability.

Quality versus quantity: Foraging decisions in the honeybee (Apis mellifera scutellata) feeding on wildflower nectar and fruit juice.

Foraging animals must often decide among resources which vary in quality and quantity. Nectar is a resource that exists along a continuum of quality in terms of sugar concentration and is the primary energy source for bees. Alternative sugar sources exist, including fruit juice, which generally has lower energetic value than nectar. We observed many honeybees (Apis mellifera scutellata) foraging on juice from fallen guava (Psidium guajava) fruit near others foraging on nectar. To investigate whether fruit and nectar offered contrasting benefits of quality and quantity, we compared honeybee foraging performance on P. guajava fruit versus two wildflowers growing within 50 m, Richardia brasiliensis and Tridax procumbens. Bees gained weight significantly faster on fruit, 2.72 mg/min, than on either flower (0.17 and 0.12 mg/min, respectively). However, the crop sugar concentration of fruit foragers was significantly lower than for either flower (12.4% vs. 37.0% and 22.7%, respectively). Fruit foragers also spent the most time handling and the least time flying, suggesting that fruit juice was energetically inexpensive to collect. We interpret honeybee foraging decisions in the context of existing foraging models and consider how nest-patch distance may be a key factor for central place foragers choosing between resources of contrasting quality and quantity. We also discuss how dilute solutions, such as fruit juice, can help maintain colony sugar-water balance. These results show the benefits of feeding on resources with contrasting quality and quantity and that even low-quality resources have value.

Appetite for self-destruction: suicidal biting as a nest defense strategy in Trigona stingless bees.

Self-sacrificial behavior represents an extreme and relatively uncommon form of altruism in worker insects. It can occur, however, when inclusive fitness benefits are high, such as when defending the nest. We studied nest defense behaviors in stingless bees, which live in eusocial colonies subject to predation. We introduced a target flag to nest entrances to elicit defensive responses and quantified four measures of defensivity in 12 stingless bee species in São Paulo State, Brazil. These included three Trigona species, which are locally known for their aggression. Species varied significantly in their attack probability (cross species range = 0-1, P < 0.001), attack latency (7.0-23.5 s, P = 0.002), biting duration of individual bees (3.5-508.7 s, P < 0.001), and number of attackers (1.0-10.8, P < 0.001). A "suicide" bioassay on the six most aggressive species determined the proportion of workers willing to suffer fatal damage rather than disengage from an intruder. All six species had at least some suicidal individuals (7-83 %, P < 0.001), reaching 83 % in Trigona hyalinata. Biting pain was positively correlated with an index of overall aggression (P = 0.002). Microscopic examination revealed that all three Trigona species had five sharp teeth per mandible, a possible defensive adaptation and cause of increased pain. Suicidal defense via biting is a new example of self-sacrificial altruism and has both parallels and differences with other self-sacrificial worker insects, such as the honey bee. Our results indicate that suicidal biting may be a widespread defense strategy in stingless bees, but it is not universal.

Monogamy in large bee societies: a stingless paradox.

High genetic diversity is important for the functioning of large insect societies. Across the social Hymenoptera (ants, bees, and wasps), species with the largest colonies tend to have a high colony-level genetic diversity resulting from multiple queens (polygyny) or queens that mate with multiple males (polyandry). Here we studied the genetic structure of Trigona spinipes, a stingless bee species with colonies an order of magnitude larger than those of polyandrous honeybees. Genotypes of adult workers and pupae from 43 nests distributed across three Brazilian biomes showed that T. spinipes colonies are usually headed by one singly mated queen. Apart from revealing a notable exception from the general incidence of high genetic diversity in large insect societies, our results reinforce previous findings suggesting the absence of polyandry in stingless bees and provide evidence against the sperm limitation hypothesis for the evolution of polyandry. Stingless bee species with large colonies, such as T. spinipes, thus seem promising study models to unravel alternative mechanisms to increase genetic diversity within colonies or understand the adaptive value of low genetic diversity in large insect societies.

Towards greater realism in inclusive fitness models: the case of worker reproduction in insect societies.

The conflicts over sex allocation and male production in insect societies have long served as an important test bed for Hamilton's theory of inclusive fitness, but have for the most part been considered separately. Here, we develop new coevolutionary models to examine the interaction between these two conflicts and demonstrate that sex ratio and colony productivity costs of worker reproduction can lead to vastly different outcomes even in species that show no variation in their relatedness structure. Empirical data on worker-produced males in eight species of Melipona bees support the predictions from a model that takes into account the demographic details of colony growth and reproduction. Overall, these models contribute significantly to explaining behavioural variation that previous theories could not account for.

Bourgeois behavior and freeloading in the colonial orb web spider Parawixia bistriata (Araneae, Araneidae).

Spiders of the tropical American colonial orb weaver Parawixia bistriata form a communal bivouac in daytime. At sunset, they leave the bivouac and construct individual, defended webs within a large, communally built scaffolding of permanent, thick silk lines between trees and bushes. Once spiders started building a web, they repelled other spiders walking on nearby scaffolding with a "bounce" behavior. In nearly all cases (93%), this resulted in the intruder leaving without a fight, akin to the "bourgeois strategy," in which residents win and intruders retreat without escalated contests. However, a few spiders (6.5%) did not build a web due to lack of available space. Webless spiders were less likely to leave when bounced (only 42% left) and instead attempted to "freeload," awaiting the capture of prey items in nearby webs. Our simple model shows that webless spiders should change their strategy from bourgeois to freeloading satellite as potential web sites become increasingly occupied.

Factors influencing survival duration and choice of virgin queens in the stingless bee Melipona quadrifasciata.

In Melipona quadrifasciata, about 10% of the females develop into queens, almost all of which are killed. Occasionally, a new queen replaces or supersedes the mother queen or heads a new colony. We investigated virgin queen fate in queenright and queenless colonies to determine the effects of queen behaviour, body mass, nestmate or non-nestmate status, queenright or queenless colony status, and, when queenless, the effect of the time a colony had been queenless, on survival duration and acceptance. None of 220 virgin queens observed in four observation hives ever attacked another virgin queen nor did any of 88 virgin queens introduced into queenright colonies ever attack the resident queen. A new queen was only accepted in a queenless colony. Factors increasing survival duration and acceptance of virgin queens were to emerge from its cell at 2 h of queenlessness, to hide, and to avoid fights with workers. In this way, a virgin queen was more likely to be available when a colony chooses a new queen, 24-48 h after resident queen removal. Running, walking or resting, antennating or trophallaxis, played little or no role, as did the factors body mass or nestmate. "Queen choice" took about 2 h during which time other virgin queens were still being killed by workers. During this agitated process, the bees congregated around the new queen. She inflated her abdomen and some of the workers deposited a substance on internal nest surfaces including the glass lid of the observation hive.

Intraspecific queen parasitism in a highly eusocial bee.

Insect societies are well-known for their advanced cooperation, but their colonies are also vulnerable to reproductive parasitism. Here, we present a novel example of an intraspecific social parasitism in a highly eusocial bee, the stingless bee Melipona scutellaris. In particular, we provide genetic evidence which shows that, upon loss of the mother queen, many colonies are invaded by unrelated queens that fly in from unrelated hives nearby. The reasons for the occurrence of this surprising form of social parasitism may be linked to the fact that unlike honeybees, Melipona bees produce new queens in great excess of colony needs, and that this exerts much greater selection on queens to seek alternative reproductive options, such as by taking over other nests. Overall, our results are the first to demonstrate that queens in highly eusocial bees can found colonies not only via supersedure or swarming, but also by infiltrating and taking over other unrelated nests.