Colony pollen reserves affect body size,sperm production and sexual development in males of the stingless bee <Emphasis Type="Italic">Melipona beecheii</Emphasis>

The production of male sexual offspring by social insect colonies is often strongly seasonal or resource-dependent. In stingless bees, males are produced in smaller numbers under conditions of low colony food reserves; whether such males are negatively affected in traits related to reproductive success is not known. We compared body size, sperm production and sexual maturity in Melipona beecheii males reared with experimentally supplemented or reduced pollen reserves, but with otherwise equal numbers of workers and equal quantities of honey reserves. We also studied the same traits in males collected from non-manipulated colonies with pollen reserves intermediate between the supplemented or reduced groups but with more workers and honey reserves. Males reared under experimentally reduced pollen reserves had significantly smaller bodies and lower sperm counts compared to those reared in colonies with experimentally supplemented pollen reserves. There was also a significantly positive relationship between the number of sperm and body size in males across all colony treatments. The maximum number of sperm in seminal vesicles was recorded 2 days later in males from colonies with reduced pollen compared to males from colonies with supplementary pollen. Males from non-manipulated colonies were intermediate in size, sperm count and speed of maturation. Our study documents for the first time the existence of large size variation in males of stingless bees that is related with the amount of pollen reserves in their natal colony. We conclude that a colony’s pollen reserves have a major impact on male body size, sperm production and speed of sexual maturity in this stingless bee, which may be the case in other social insects. Stingless bees are a good model system to study the balance between colony-level selection and individual-level selection on male sexually selected traits such as body size.     

Ontogeny of worker body size distribution in bumble bee (Bombus impatiens) colonies

1. Bumble bees exhibit worker size polymorphisms; highly related workers within a colony may vary up to 10‐fold in body mass. As size variation is an important life history feature in bumble bees, the distribution of body sizes within the colony and how it fluctuates over the colony cycle were analysed. 2. Ten commercially purchased colonies of Bombus impatiens (Cresson) were reared in ad libitum conditions. The size of all workers present and newly emerging workers (callows) was recorded each week. 3. The average size of bumble bee workers did not change with colony age, but variation in body size tended to decrease over time. The average size of callows did not change with population size, but did tend to decrease with colony age. In all measures, there was considerable variation among colonies. 4. Colonies of B. impatiens usually produced workers with normally distributed body sizes throughout the colony life cycle. Unlike most polymorphic ants, there was no increase in worker body size with colony age or colony size. This provides the first, quantitative data on the ontogeny of bumble bee worker size distribution. The potential adaptive significance of this size variation is discussed.     

Imidacloprid-induced impairment of mushroom bodies and behavior of the native stingless bee Melipona quadrifasciata anthidioides

Declines in pollinator colonies represent a worldwide concern. The widespread use of agricultural pesticides is recognized as a potential cause of these declines. Previous studies have examined the effects of neonicotinoid insecticides such as imidacloprid on pollinator colonies, but these investigations have mainly focused on adult honey bees. Native stingless bees (Hymenoptera: Apidae: Meliponinae) are key pollinators in neotropical areas and are threatened with extinction due to deforestation and pesticide use. Few studies have directly investigated the effects of pesticides on these pollinators. Furthermore, the existing impact studies did not address the issue of larval ingestion of contaminated pollen and nectar, which could potentially have dire consequences for the colony. Here, we assessed the effects of imidacloprid ingestion by stingless bee larvae on their survival, development, neuromorphology and adult walking behavior. Increasing doses of imidacloprid were added to the diet provided to individual worker larvae of the stingless bee Melipona quadrifasciata anthidioides throughout their development. Survival rates above 50% were only observed at insecticide doses lower than 0.0056 μg active ingredient (a.i.)/bee. No sublethal effect on body mass or developmental time was observed in the surviving insects, but the pesticide treatment negatively affected the development of mushroom bodies in the brain and impaired the walking behavior of newly emerged adult workers. Therefore, stingless bee larvae are particularly susceptible to imidacloprid, as it caused both high mortality and sublethal effects that impaired brain development and compromised mobility at the young adult stage. These findings demonstrate the lethal effects of imidacloprid on native stingless bees and provide evidence of novel serious sublethal effects that may compromise colony survival. The ecological and economic importance of neotropical stingless bees as pollinators, their susceptibility to insecticides and the vulnerability of their larvae to insecticide exposure emphasize the importance of studying these species.     

Key factors influencing forager distribution across macadamia orchards differ among species of managed bees

To achieve maximised and sustainable crop productivity, it is critical that we develop crop-specific strategies for managing pollination. Honey bees (Apis mellifera) and stingless bees (Tetragonula carbonaria) are considered effective pollinators of macadamia (Macadamia integrifolia). The introduction of managed honey bee or stingless bee hives into orchards is likely to boost the numbers of these insects visiting flowers; however, there is a lack of published information and consensus regarding their management for pollination. Here, we identify factors that affect the distribution of both honey bees and stingless bees across cultivated macadamia, and establish whether increased flower visitation leads to higher nut set. A gradient of bee visitation rates was created by placing colonies on the ends of a four-hectare block, and mixed-effect models were applied to assess forager abundance and nut set with respect to distance from hive, time of day, cultivar, and floral display size. Distance from colony had a strong effect on stingless bee numbers, with >96% of individuals recorded within 100 metres of colonies, whereas the distribution of honey bees was more closely related to daily floral display: trees with greater numbers of flowers attracted more honey bees. Simplified surveys conducted in a further 17 macadamia blocks confirm that these are broadly occurring distribution patterns. Bee abundance alone did not significantly predict nut production; however, an indirect effect of bee visits to flowers is inferred, as nut production increased with size of floral display. To encourage a more even distribution of bees and uniform pollination, we recommend placement of stingless bee hives to maximise their distribution through a block (e.g. at 100-m intervals) and management practices that promote even distributions of flowers across trees.     

Does intraspecific size variation in bumblebees allow colonies to efficiently exploit different flowers?

Abstract.  1. It has long been known that foraging bumblebee workers vary greatly in size, within species, and within single nests. This phenomenon has not been adequately explained. Workers of their relatives within the Apidae exhibit much less size variation.
2. For the bumblebee Bombus terrestris size, as measured by thorax width, was found to correspond closely with tongue length, so that larger bees are equipped to feed from deeper flowers.
3. The mean size of worker bees attracted to flowers was found to differ between plant species, and larger bees with longer tongues tended to visit deeper flowers.
4. Finally, handling time depended on the match between corolla depth and tongue length: large bees were slower than small bees when handling shallow flowers, but quicker than small bees when handling deep flowers.
5. Size variation within bumblebees may be adaptive, since it enables the colony as a whole to efficiently exploit a range of different flowers. Possible explanations for the marked differences in size variation exhibited by bumblebees compared with Apis species and stingless bees (Meliponinae) are discussed.     

ADAPTIVE GENETIC VARIATION IN GROWTH AND DEVELOPMENT OF TADPOLES OF THE HYBRIDOGENETIC RANA ESCULENTA COMPLEX

The distribution and proportion of the sexual species Rana lessonae to the hemiclonal hybrid R. esculenta among natural habitats suggests that these anurans may differ in adaptive abilities. I used a half-sib design to partition phenotypic and quantitative genetic variation in tadpole responses at two food levels into causal variance components. Rana lessonae displays strong phenotypic variation across food levels. Growth rate is strictly determined by environmental factors and includes weak maternal effects. Larval period and body size at metamorphosis both contain moderate levels of additive genetic variance. The sire x food interactions and the lack of environmental correlations indicate that adaptive phenotypic plasticity is present in both of these traits. In contrast, R. esculenta displays less phenotypic variation across food levels, especially for larval period. Variation in body size at metamorphosis is underlain by genetic variation as shown by high levels of additive genetic variance, yet growth rate and larval period are not. Significant environmental correlations between larval period at high food level and growth, larval period, and body size at low food, indicate phenotypic plasticity is absent. A positive phenotypic correlation between body size at metamorphosis and larval period for R. lessonae at both food levels suggests a trade-off between growing large and metamorphosing quickly to escape predation or pond drying. The lack of a similar correlation for R. esculenta at the high food level suggests it may be less constrained. Different levels of adaptive genetic variation among larval traits suggest that the sexual species and the hybridogenetic hemiclone differ in their abilities to cope with temporally and spatially heterogeneous environments.     

Thermal constraints for stingless bee foragers: the importance of body size and coloration

In the dry tropics, foraging bees face significant thermal constraints as a result of high ambient temperatures and direct insolation. In order to determine the potential importance of body size and body coloration in heat gain and heat loss, passive warm-up and cooling rates were measured for freshly killed workers of 24 stingless bee species. Results accorded with biophysical principles. Small bees reached lower temperature excesses (T_exc) and warmed up and lost heat much more rapidly than larger bees. In addition to body size, body coloration had a clear effect on thermal parameters. Light-coloured bees warmed up less rapidly and had lower T_exc than dark bees. An intraspecific comparison of Melipona costaricensis and Cephalotrigona capitata colour morphs confirmed that body coloration influences thermal characteristics. This study is the first to indicate that abdominal coloration in stingless bees might be involved in the regulation of body temperature in extreme thermal conditions. However, body temperatures of foraging bees of colour morphs were not very different. This is probably due to behavioural adaptations (e.g. foraging strategies) or differences in convective and evaporative heat loss or the production of metabolic heat during flight, that all mask the effect of body colour. Notwithstanding such effects and potential thermoregulatory capabilities, stingless bees show niche differentiation and biogeographic distributions that correlate with body coloration and body size. This also suggests that, in general, light bees have an advantage over black bees in hot open lowland habitats, whereas black bees might have an advantage in wet habitats and mountains. The origin, occurrence and function of flavinism (yellow integument colouring) are discussed.     

An exploration of the relationship between recruitment communication and foraging in stingless bees

Social information is widely used in the animal kingdom and can be highly adaptive. In social insects, foragers can use social information to find food, avoid danger, or choose a new nest site. Copying others allows individuals to obtain information without having to sample the environment. When foragers communicate information they will often only advertise high-quality food sources, thereby filtering out less adaptive information. Stingless bees, a large pantropical group of highly eusocial bees, face intense inter- and intra-specific competition for limited resources, yet display disparate foraging strategies. Within the same environment there are species that communicate the location of food resources to nest-mates and species that do not. Our current understanding of why some species communicate foraging sites while others do not is limited. Studying freely foraging colonies of several co-existing stingless bee species in Brazil, we investigated if recruitment to specific food locations is linked to 1) the sugar content of forage, 2) the duration of foraging trips, and 3) the variation in activity of a colony from 1 day to another and the variation in activity in a species over a day. We found that, contrary to our expectations, species with recruitment communication did not return with higher quality forage than species that do not recruit nestmates. Furthermore, foragers from recruiting species did not have shorter foraging trip durations than those from weakly recruiting species. Given the intense inter- and intraspecific competition for resources in these environments, it may be that recruiting species favor food resources that can be monopolized by the colony rather than food sources that offer high-quality rewards.     

Ergonomics in stingless bees: changes in intranidal behavior after partial removal of storage pots and honey inMelipona favosa (Hym. Apidae,Meliponinae)

Summary The ergonomics of task allocation in stingless bees was examined in two laboratory colonies ofMelipona favosa containing individually marked workers. Performance of intranidal behavior patterns, brood cell production and the processing and storage of nectar, were studied during a control and a wax-deprived period. Experimental wax deprivation consisted of regular controlled removal of storage pots and their contents. Stored carbohydrate concentration and storage pot mass declined during the wax-deprived period. Behavior was measured by sampling for a complete ethogram at seven intranidal areas.In comparison withApis mellifera, M. favosa appeared to be more sensitive to stress, with a distinct reallocation of effort between tasks. The significant increase under stress of brood production and brood-rearing behavior patterns observed was previously also found inApis. Self-oriented behavior patterns declined under stressed circumstances.Involucrum construction declined in wax-deprivedMelipona colonies. Relatively low rates of walking behavior in general inMelipona suggest a low mean free distance between tasks. These results are of particular interest in relation to the basically different nest structure of stingless bees, e.g. very distinct areas for brood production and the processing and storage of food. This architecture of stingless bee nests appeared to influence strongly their specific responses to wax deprivation.     

The cuticular profiles of Australian stingless bees are shaped by resin of the eucalypt tree Corymbia torelliana

Bees are known to collect pollen and nectar to provide their larvae and themselves with food. That bees, especially the tropical stingless bees (Apidae: Meliponini), also collect plant resins has, however, been barely addressed in scientific studies on resource use in bees. Resins are used for nest construction, nest maintenance and nest defence. Furthermore, some South‐East Asian species transfer resin‐derived terpenes to their cuticular profiles. The resin requirement of bees is in turn used by certain plant species, which attract bees either for pollination by providing resin in their inflorescences, or for seed dispersal by providing resin in their seed capsules (mellitochory). Mellitochory is found in the eucalypt tree Corymbia torelliana, the resin of which is collected by Australian stingless bees. We investigated how the interaction between C. torelliana and resin‐collecting bees affects the chemical ecology of two Australian stingless bee genera by comparing the chemical profiles of eight bee species with resin from C. torelliana fruits. The two bee genera differed significantly in their chemical profiles. Similar to South‐East Asian stingless bees, 51% of all compounds on the body surfaces of the five Tetragonula species were most likely derived from plant resins. Up to 32 compounds were identical with compounds from C. torelliana resin, suggesting that Tetragonula species include C. torelliana compounds in their chemical profiles. By contrast, few or none resinous compounds were found on the body surfaces of the three Austroplebeia species sampled. However, one prominent but as yet unknown substance was found in both C. torelliana resin and the chemical profiles of all Tetragonula and four Austroplebeia colonies sampled, suggesting that most colonies (76%) gathered resin from C. torelliana. Hence, C. torelliana resin may be commonly collected by Australian stingless bees and, along with resins from other plant species, shape their chemical ecology.     

Oviposition behavior and related notes of the Taiwanese stingless bee

Behavioral observations ofTrigona (Lepidotrigona) ventralis hoozana revealed the following ethological features: (1) Cell construction and food provisioning in cells are semisynchronous, and ovipositions are always batched. (2) The queen displays a peculiar behavior, “repeated turnings”, in front of the provisioned cells. (3) The queen often violently solicits workers food and, unlike other studied taxa, workers feed the queen rather frequently. (4) Worker oviposition is frequent when the colony is queen-right. An ethological diagnosis ofLepidotrigona is given with comparisons to other stingless bee taxa. Contribution to the knowledge of the Indo-Pacific stingless bees. IX. Behavior studies of the stingless bees, with special reference to the oviposition process. XII.     

Genetic structure of nest aggregations and drone congregations of the southeast Asian stingless bee Trigona collina

In stingless bees, sex is determined by a single complementary sex-determining locus. This method of sex determination imposes a severe cost of inbreeding because an egg fertilized by sperm carrying the same sex allele as the egg results in a sterile diploid male. To explore how reproductive strategies may be used to avoid inbreeding in stingless bees, we studied the genetic structure of a population of 27 colonies and three drone congregations of Trigona collina in Chanthaburi, Thailand. The colonies were distributed across six nest aggregations, each aggregation located in the base of a different fig tree. Genetic analysis at eight microsatellite loci showed that colonies within aggregations were not related. Samples taken from three drone congregations showed that the males were drawn from a large number of colonies (estimated to be 132 different colonies in our largest swarm). No drone had a genotype indicating that it could have originated from the colony that it was directly outside. Combined, these results suggest that movements of drones and possibly movements of reproductive swarms among colony aggregations provide two mechanisms of inbreeding avoidance.     

Adaptation of hypopharyngeal gland development to the brood status of honeybee (Apis mellifera L.) colonies

We studied pollen consumption, head weight, hypopharyngeal gland (HPG) acini diameter, and protein synthesis and transfer in honeybee workers reared in colonies with normal and with decreasing amounts of brood. We found that head fresh weight is correlated with size of the glands and that pollen consumption is positively correlated with gland development. An effect of brood on size of the glands could be confirmed, but was not as profound as in previous studies. Similarly, no difference in the amount of protein synthesized or transferred in workers living under the two brood conditions was found. We suspect this is due to the fact that HPGs also supply food to young bees and in our study young bees were always present while in previous studies, colonies often lacked both brood and young bees.     

Natural patterns of caste and sex allocation in the stingless bees <Emphasis Type="Italic">Melipona favosa</Emphasis> and <Emphasis Type="Italic">M. trinitatis</Emphasis> related to worker behaviour

Summary. This paper is the report of a long-duration study of the production of gynes and males in Melipona stingless bees in Trinidad and Tobago, including a behavioural study of laying workers. In 167 brood samples (16342 pupae) of M. favosa, taken from 78 colonies in the natural environment over the years 1993-2001, workers represented 78.4% and males represented 17.3%. Gynes represented 5.1% of all female brood. Gyne proportion never exceeded 22.5%. Gynes were found to be common throughout the year. Like the gynes, males were present all year, but their occurrence varied over the months. Males were most common in the months of July and August. At the colony level male occurrence in the brood was characterized by very high values (maximum 74.2%), next to a frequent complete absence (in 26.5% of all samples). The finding of bout-like production of males at the colony level is supported by the observations on reproductive worker behaviour. The colonies appeared to be in different phases of male production. When male producing laying workers were present, they competed heavily for egg-laying opportunities. Various behaviours of laying workers and the queen are described and interpreted on the basis of reproductive competition and kin conflict theories. From these results, we conclude that in M. favosa, workers have a distinct influence on reproduction, including the production of gynes and males.     

Experimentally induced variation in the physical reproductive potential and mating success in honey bee queens

In honeybee colonies, reproduction is monopolized by the queen while her daughter workers are facultatively sterile. Caste determination is a consequence of environmental conditions during development, during which female larvae may become either queens or workers depending on their larval diet. This bipotency introduces significant variation in the reproductive potential of queen bees, with queens raised from young worker larvae exhibiting high reproductive potential and queens raised from older worker larvae exhibiting lower reproductive potential. We verify that low-quality queens are indeed produced from older worker larvae, as measured morphometrically (e.g., body size) and by stored sperm counts. We also show, for the first time, that low-quality queens mate with significantly fewer males, which significantly influences the resultant intracolony genetic diversity of the worker force of their future colonies. These results demonstrate a reproductive continuum of honeybee queens and provide insights into the reproductive constraints of social insects.     

Mate number,kin selection and social conflicts in stingless bees and honeybees

Microsatellite genotyping of workers from 13 species (ten genera) of stingless bees shows that genetic relatedness is very high. Workers are usually daughters of a single, singly mated queen. This observation, coupled with the multiple mating of honeybee queens, permits kin selection theory to account for many differences in the social biology of the two taxa. First, in contrast to honeybees, where workers are predicted to and do police each other''s male production, stingless bee workers are predicted to compete directly with the queen for rights to produce males. This leads to behavioural and reproductive conflict during oviposition. Second, the risk that a daughter queen will attack the mother queen is higher in honeybees, as is the cost of such an attack to workers. This explains why stingless bees commonly have virgin queens in the nest, but honeybees do not. It also explains why in honeybees the mother queen leaves to found a new nest, while in stingless bees it is the daughter queen who leaves.     

Outbreeding and lack of temporal genetic structure in a drone congregation of the neotropical stingless bee Scaptotrigona mexicana

Drone aggregations are a widespread phenomenon in many stingless bee species (Meliponini), but the ultimate and proximate causes for their formation are still not well understood. One adaptive explanation for this phenomenon is the avoidance of inbreeding, which is especially detrimental for stingless bees due to the combined effects of the complementary sex-determining system and the small effective population size caused by eusociality and monandry. We analyzed the temporal genetic dynamics of a drone aggregation of the stingless bee Scaptotrigona mexicana with microsatellite markers over a time window of four weeks. We estimated the drones of the aggregation to originate from a total of 55 colonies using sibship re-construction. There was no detectable temporal genetic differentiation or sub-structuring in the aggregation. Most important, we could exclude all colonies in close proximity of the aggregation as origin of the drones in the aggregation, implicating that they originate from more distant colonies. We conclude that the diverse genetic composition and the distant origin of the drones of the S. mexicana drone congregation provides an effective mechanism to avoid mating among close relatives.     

Rates of juvenile hormone synthesis control caste differentiation in the stingless bee Scaptotrigona postica depilis

Summary In social insects the expression of caste-specific characters is controlled by juvenile hormone (JH) during definite sensitive periods in preimaginal development. For a number of stingless bee species the existence of such a JH-sensitive period has already been demonstrated. Queen development can be induced by topical JH applications during the cocoon spinning phase of the last larval instar. Neither JH titers nor rates of JH synthesis were known so far for this subfamily of eusocial bees distinguished by a pronounced caste dimorphism. As the pantropically distributed stingless bees with approximately 400 recent species are the largest group of social bees, JH synthesis was studied in one of the species that can be kept under laboratory conditions. An in vitro radiochemical assay was used to measure stage- and caste-specific activities of the corpora allata (CA). For the first time in a eusocial hymenopteran species it was demonstrated how the endocrine system is reacting to trophogenic stimuli capable to induce caste differentiation during larval development. Generally JH synthesis in queen CA was found to be 30–80% higher than in workers during the penultimate and last larval instar, but a strong and distinct caste-specific modulation of JH synthesis was only observed right before the onset of a JH-sensitive period in the cocoon spinning phase of the fifth instar.     

Clustered male production by workers in the stingless bee Melipona subnitida Ducke (Apidae, Meliponinae)

Summary: In stingless bees brood cells are sequentially filled with liquid larval food (mass-provisioning), upon which the queen lays an egg. Thereafter the cell is closed by a worker. This study showed that during these processes workers of Melipona subnitida regularly laid eggs that served as food for the queen. Occasionally cells were oviposited in and immediately closed by a worker. These cells always rendered males. Some of these reproductive workers were seen to lay a trophic egg as well. Cells which were exclusively oviposited in by the physogastric queen gave rise to workers and queens only. In one colony it could be verified that three workers alone, which differed in age by one day, laid 15 male-producing eggs within a period of two successive weeks. Among them the number of ovipositions was positively related to the order in which workers eclosed - the oldest worker laying most eggs - and inversely related to the number of times they closed cells oviposited in exclusively by the queen. Apparently the physogastric queen was not able to stop certain workers from reproducing. We therefore conclude that some workers in M. subnitida temporarily dominated their queens in egg-laying.     

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.