Sex ratio variation in the bumblebee Bombus terrestris

Patterns of sex allocation in bumblebees have been enigmaticand difficult to interpret in either a Fisherian context orin a kin-selection perspective. We gathered data on severalhundred laboratory-reared colonies of the bumblebee Bombus terrestrisand analyzed sex allocation as a function of diapause durationand a series of variables describing colony development. Ouranalyses addressed both sex allocation patterns across differentcohorts of laboratory colonies reared at different times andsex allocation patterns across individual colonies within thesecohorts. We used path analysis to test a hypothetical modellinking a sequence of colony-development variables to the crucialreproductive parameters at the end of the colony life cycle.We show that (1) population-wide patterns of sex allocationshow equal investment in the sexes and are thus consistent withqueen control, but not with worker control. (2) A significantpart of the colony-level and cohort-specific variation in sexallocation is related to the hibernation conditions of foundingqueens: Queens with longer than average winter diapause producelarger cohorts of first and second brood workers, switch tohaploid eggs early, and produce colonies that raise mostly malesand few new queens and vice versa. (3) Colony-level sex allocationis significantly related to the time span between the switchpoint (date of first haploid egg laid by the queen) and thecompetition point (date of first haploid egg laid by one ofthe workers): the longer this period, the more male biased thesex ratio. (4) The breeding constraints of an annual life cycle,the short reproductive season, and the presumably high premiumon early produced males imply that bumblebee workers have norealistic options to capitalize on their relatedness asymmetrytoward the different kinds of reproductive brood by biasingthe sex ratio.     

Respiration and energetics of the bumblebee Bombus terrestris queen

Carbon dioxide production by the Bombus terrestris queen was measured at different temperatures (10–30°C) and during different activities of the bumblebee. During flight the CO₂ production averaged 50 ml g⁻¹ (fresh weight) h⁻¹ and was only slightly affected by temperature. During rest (with a readiness to fly) and incubation the respiration rate clearly increased with decreasing temperature (5–40 and 13–56 ml g⁻¹ h⁻¹, respectively), whilst during torpor it increased with temperature (0.1–1.7 ml g⁻¹ h⁻¹ at temperatures from 10 to 30°C).
The expenditure of energy as calculated from the continuous respiration measurements agreed well with the amount of energy obtained from food (discrepancy 6–19%). The energy budget of an incubating queen was correctly predicted using the measured respiratory functions, prevailing temperatures, and the behaviour of the queen. The number of flower visits needed to fulfil the daily energy requirements of an incubating queen is discussed.     

A larval hunger signal in the bumblebee Bombus terrestris

Larvae of Bombus terrestris, a pollen-storing bumblebee, are dependent on progressive provisioning by workers. We test the hypothesis that larval cuticular chemicals can act as a hunger signal. We first show with a new classical conditioning experiment, using a Y-shaped tube, that workers can be trained to prefer the extracts of normally fed larvae over those of starved larvae. This proves the ability of workers to discriminate between larval extracts. Second, we show in a bioassay that workers also use these perceived differences to feed larvae according to their nutritional status. Larval broods sprayed with the extracts of the starved larvae were fed significantly more than larval broods sprayed with the extracts of normally fed larvae or with the solvent (n-pentane) only. We therefore conclude that B. terrestris larvae signal their need for food via their cuticular chemicals, and discuss the extent to which this form of communication could give larvae some control over their development. Received 2 September 2005; revised 11 April; accepted 24 April 2006.     

Behavioural interactions, kin and disease susceptibility in the bumblebee Bombus terrestris

Behavioural interactions are often analysed in terms of their costs and benefits to the actors [Hamilton, (1964) J. Theor. Biol.7 1-16; Gadagkar, (1993) Trends Ecol. Evol.8 232-234; Foster et al., (2001) Ann. Zool. Fenn.38 229-238]. Using the bumblebee Bombus terrestris, we wish to distinguish between two possible determinants of interaction behaviour between conspecifics, namely kin-directed behaviour that reflects genetic distance between individuals, or, alternatively, interactions guided by a functional distance between individuals, specifically, with respect to disease susceptibility. We find no relationship between contact rate of individuals and the genetic distance of their respective colonies. Interestingly, we do find a significant negative correlation between contact rate and the distance between the two colonies in susceptibility to a spectrum of parasite strains. This cannot be explained by either of the a priori alternatives so we propose two further testable hypotheses to explain our results.     

Isolation and characterization of chitin from bumblebee (Bombus terrestris)

Insect chitin possessing shell-like structure was prepared from the bumblebee corpses by a consequent treatment with 1M HCl and 1M NaOH. The bumblebee chitin was compared with crustacean (shrimp) chitin by using elemental analysis, Fourier-transform infrared (FT-IR) and solid-state (13)C cross-polarization magic angle spinning nuclear magnetic resonance (CP/MAS)-NMR spectroscopy and confocal microscopy. Both chitins (bumblebee and shrimp) exhibited identical spectra, while the bumblebee chitin had a 5% lower degree of acetylation and was characterized by a fine membrane texture.     

Caste-specific differences in ecdysteroid titers in early larval stages of the bumblebee Bombus terrestris

Mounting evidence implicates ecdysteroids in queen-worker differentiation during the last larval instars of highly social insects. In the present study, we analyzed ecdysteroid titers in queen and worker larvae of the bumblebee Bombus terrestris from the second to the early fourth instar. B. terrestris is of particular interest because caste is already determined in the second instar, presumably by a pheromonal signal emitted by the egg-laying queen. Caste differences in the adults, however, are only expressed at the physiological and not at the morphological level, except for the distinctly larger size of the queen. In the second and third instar, ecdysteroid titers in queen larvae were generally higher than those of workers. These early caste-specific differences, however, were abolished in the fourth instar. In the early fourth instar we could detect two small ecdysteroid peaks, with the one preceding the cocoon-spinning phase presenting the characteristics of a pupal commitment peak. The synchrony of caste differences in ecdysteroid and juvenile hormone titers suggests a synergistic action of these hormones in caste determination.     

The composition of larval food and the significance of exocrine secretions in the bumblebee Bombus terrestris

Summary: This paper describes a study on the relation between the composition of larval food and the development of female castes in bumblebees. The first aim was to evaluate the significance of glandular secretions in the larval diet as a possible factor involved in larval feeding and caste differentiation. Small amounts of proteinaceous secretions were found to be added during the ingestion of sucrose but not while discharging food to the larvae. It is discussed that these secretions are digestive in function rather than food additives that would possibly play a role in the process of caste differentiation.¶Secondly, a comparative analysis was made of the general composition of food samples obtained from larvae of different castes and ages and from various periods in the social development of the colony. No significant differences in the total amount of pollen, sucrose and protein were detected between the castes or different age groups. Unlike honeybee workers, individual bumblebee workers did not change the composition of the diet they supplied to the brood in relation to their own age, nor to the social development of the colony. These findings suggest that all larvae receive the same nourishment during their total development and indicate that differences in development between queen larvae and worker larvae are neither caused by variations nor by a qualitative modification of their food with respect to the amount of pollen, protein and carbohydrates.     

Natural variation in the genetic architecture of a host-parasite interaction in the bumblebee Bombus terrestris

The genetic architecture of fitness-relevant traits in natural populations is a topic that has remained almost untouched by quantitative genetics. Given the importance of parasitism for the host's fitness, we used QTL mapping to study the genetic architecture of traits relevant for host-parasite interactions in the trypanosome parasite, Crithidia bombi and its host, Bombus terrestris. The three traits analysed were the parasite's infection intensity, the strength of the general immune response (measured as the encapsulation of a novel antigen) and body size. The genetic architecture of these traits was examined in three natural, unmanipulated mapping populations of B. terrestris. Our results indicate that the intracolonial phenotypic variation of all three traits is based on a network of QTLs and epistatic interactions. While these networks are similar between mapping populations in complexity and number of QTLs, as well as in their epistatic interactions, the variability in the position of QTL and the interacting loci was high. Only one QTL for body size was plausibly found in at least two populations. QTLs for encapsulation and Crithidia infection intensity were located on the same linkage groups.     

An immune response in the bumblebee, Bombus terrestris leads to increased food consumption

Background  

The concept of a costly immune system that must be traded off against other important physiological systems is fundamental to the burgeoning field of ecological immunity. Bumblebees have become one of the central models in this field. Although previous work has demonstrated costs of immunity in numerous life history traits, estimates of the more direct costs of bumblebee immunity have yet to be made.     

Photoperiodic influence on the body mass of bumblebee, Bombus terrestris and its copulation duration

Abstract:  The body mass of Bombus terrestris individuals is an important trait for their behavioural performance and colony organization. In this study, colonies were reared under four different photoperiodic regimes, viz. 0 : 24, 8 : 16, 16 : 8 and 24 : 0 h light : darkness (L : D) at 28°C and 50% relative humidity. The changes in body mass were observed at the stages of larvae, pupae and on the day of adult eclosion. Both the wet and dry mass of sexuals gradually decreased with increasing day length. The relationship between body mass and copulation duration revealed that copulation duration was negatively correlated with male body mass, but positively with queen body mass. Higher number of matings by males resulted in significantly higher duration of copulation.     

Isolation and properties of flight muscle mitochondria of the bumblebee Bombus terrestris (L.)

This report describes the isolation procedure and properties of tightly coupled flight muscle mitochondria of the bumblebee Bombus terrestris (L.). The highest respiratory control index was observed upon oxidation of pyruvate, whereas the highest respiration rates were registered upon oxidation of a combination of the following substrates: pyruvate + malate, pyruvate + proline, or pyruvate + glutamate. The respiration rates upon oxidation of malate, glutamate, glutamate + malate, or succinate were very low. At variance with flight muscle mitochondria of a number of other insects reported earlier, B. terrestris mitochondria did not show high rates of respiration supported by oxidation of proline. The maximal respiration rates were observed upon oxidation of α-glycerophosphate. Bumblebee mitochondria are capable of maintaining high membrane potential in the absence of added respiratory substrates, which was completely dissipated by the addition of rotenone, suggesting high amount of intramitochondrial NAD-linked oxidative substrates. Pyruvate and α-glycerophosphate appear to be the optimal oxidative substrates for maintaining the high rates of oxidative metabolism of the bumblebee mitochondria.     

Effects of selective episodes in the field on life history traits in the bumblebee Bombus terrestris

Natural selection has different fitness consequences when it acts during different life cycle stages of an organism. In social insects, for example, the colony foundation and early colony growth is a critical time period with high probability of failure. Here we used colonies of the bumblebee Bombus terrestris L. to test whether selective episodes at different life cycle stages result in differences in colony performance and fitness. The timing of a selective episode was varied by field exposure of colonies, either permanently or during a short period at three different colony life cycle stages – early, middle, or late in the cycle. We found that selective episodes at different life cycle stages did not affect maximal size, fitness or survival of colonies, or the time span between colony foundation and reproduction. Instead, the colonies were able to compensate for costs encountered by delaying reproduction. This might have important fitness consequences, since later emerging sexuals might be faced with harsher environmental conditions and increased costs of finding a mate. In addition, an important component of selection might be parasitism and the resulting resource allocation to the immune system. We here measured the generalized immune response (i.e. encapsulation response) of early produced workers as an indicator of a colony's capacity to defend against parasitism. Encapsulation response correlated positively with eventual colony size and fitness, indicating that this measure of "immunocompetence" correlates with important life history traits.     

Invasion success of the bumblebee, Bombus terrestris, despite a drastic genetic bottleneck

In early 1992, the European bumblebee, Bombus terrestris, was first seen in Tasmania and currently has spread to most of the island. Here, we report on the genetic structure, using micro-satellites, of the invading population from samples collected in the years 1998-2000, a few years after the first sighting of the species in its new area. The data show that the Tasmanian population has a very low genetic diversity, with less than half of the allelic richness (Richness=2.89 alleles; H(exp)=0.591) and lower levels of heterozygosity as compared to populations in New Zealand (4.24 alleles; H(exp)=0.729) and Europe (5.08 alleles; H(exp)=0.826). In addition, the genetic data suggest that the invasion must have happened once, probably around late 1991, and was the result of very few, perhaps only two, individuals arriving in Tasmania. Furthermore, these founders came from the New Zealand population. Today, the population in the south of Tasmania seems to act as a source population from which individuals migrate into other parts of the state. A similar source-sink structure seems also the case for New Zealand. The data show that B. terrestris is a highly invasive species capable of establishing itself even after a dramatic genetic bottleneck. B. terrestris may be an invasive species due to the haplo-diploid sex determination system, which exposes recessive, deleterious mutations to selection. Offspring of such purged lines may then be able to tolerate high levels of inbreeding.     

Selection for non-diapause in the bumblebee Bombus terrestris, with notes on the effect of inbreeding

Bumblebees are generally believed to be annual insects. However, here we will show that under laboratory conditions the bumblebee Bombus terrestris (L.) can produce a second generation without a period of cold storage (diapause) or CO₂ narcosis (a method to break diapause). It is also shown that this so-called non-diapause trait can be selected for. The percentage of non-diapausing queens increased from 8% (minimum) to 97% (maximum) in two generations of selection. However, it was not possible to maintain isofemale non-diapause lines. Colonies of the fourth and fifth generation remained small (expressed in worker number) and produced a small number of queens. Also the percentage of queens that started laying eggs (defined as the percentage non-diapause) decreased in the fourth and fifth generations. To study whether this decline of the non-diapause lines was caused by inbreeding, a control experiment was conducted. In this control experiment queens were mated with their brothers (full-sib mating) for several generations and the number of queens that start egg laying was measured. This revealed that inbreeding can have a negative effect on the egg-laying capacities of queens thus causing the decline of inbred (non-diapause) lines.     

Reproductive success of a colony of the introduced bumblebee Bombus terrestris (L.) (Hymenoptera: Apidae) in a Tasmanian National Park

Abstract  Ever since a feral population of the bumblebee Bombus terrestris was found in suburban Hobart, on the Australian island of Tasmania, there has been ongoing debate surrounding the capacity of this species to utilise Australian native vegetation. Although several studies have reported B. terrestris foraging on Tasmanian native vegetation, doubts have been raised as to whether this reflects successful breeding in native vegetation. This study documents the success of a colony of B. terrestris in a Tasmanian National Park, isolated from urban and agricultural areas by 10 km of sea. Examination of the larval cocoons revealed that this colony had produced at least 304 new queens and 939 workers and drones. Pollen stores found in the colony were mostly from native plants, particularly Eucalyptus. These results strongly suggest that B. terrestris is able to reproduce successfully in parts of Australia that still support almost exclusively native vegetation.     

Membrane filtering properties of the bumblebee (Bombus terrestris) photoreceptors across three spectral classes

Filtering properties of the membrane form an integral part of the mechanisms producing the light-induced electrical signal in insect photoreceptors. Insect photoreceptors vary in response speed between different species, but recently it has also been shown that different spectral photoreceptor classes within a species possess diverse response characteristics. However, it has not been quantified what roles phototransduction and membrane properties play in such diversity. Here, we use electrophysiological methods in combination with system analysis to study whether the membrane properties could create the variation of the response speed found in the bumblebee (Bombus terrestris) photoreceptors. We recorded intracellular responses from each photoreceptor class to white noise-modulated current stimuli and defined their input resistance and linear filtering properties. We found that green sensitive cells exhibit smaller input resistance and membrane impedance than other cell classes. Since green sensitive cells are the fastest photoreceptor class in the bumblebee retina, our results suggest that the membrane filtering properties are correlated with the speed of light responses across the spectral classes. In general, our results provide a compelling example of filtering at the sensory cell level where the biophysical properties of the membrane are matched to the performance requirements set by visual ecology.     

The degree of parasitism of the bumblebee (Bombus terrestris) by cuckoo bumblebees (Bombus (Psithyrus) vestalis)

Host–parasite systems are characterised by coevolutionary arms races between host and parasite. Parasites are often the driving force, as they replicate much faster than their hosts and have shorter generation times and larger population sizes, resulting in higher mutation rates per time interval. This scenario does not fit all host–parasite systems. Socially parasitic cuckoo bumblebees (Bombus (Psithyrus) vestalis) parasitise colonies of Bombus terrestris share most life history characteristics with their hosts. As they parasitise only a subset of all available colonies, their population size should be lower than that of their hosts. This might have strong negative effects on the genetic diversity of B. vestalis and their adaptability. Here, we study for the first time the population structure of a Bombus/Bombus (Psithyrus) system. Highly polymorphic DNA markers were used to reconstruct sibships from individuals collected in the wild. The analysis of the host and parasite populations revealed a rate of parasitism of about 42% (range 33–50%). The population size of B. vestalis was lower compared to their hosts, which was also reflected in low within-group genetic distance. An analysis of the reconstructed queen genotypes revealed more supersisters amongst the B. vestalis queens when compared to the B. terrestris host. The data suggest that B. vestalis females and males do not disperse over long distances. This shows a potential for local adaptation to their hosts.     

Reproductive responses to photoperiod and temperature by artificially hibernated bumblebee (Bombus terrestris) queens

Post‐hibernated bumblebee (Bombus terrestris) queens were kept for 1 week under photoperiodic conditions of 8 h light : 16 h dark, and at four different temperatures (24, 28, 32 and 36°C). The reproductive performance of the queens was then observed. It was found that exposure temperature and hibernation duration did not affect the oviposition rate. The pre‐oviposition period was found to be shortest (3.8 ± 0.7 days) for queens that had hibernated for 4.0 months and had been activated at 28°C. Timing of the initiation of the switch‐point was not affected by exposure temperature and hibernation duration. Significantly higher numbers of workers (268.0 ± 31.4) and sexual queens (119.3 ± 16.8) were produced by the queens that had hibernated for 3.0 months and had been activated at 28 and 36°C, respectively. The queens that had hibernated for 4.0 months and had been activated at 36°C produced the highest number of males (296.2 ± 32.3).     

Construction and characterization of a BAC-library for a key pollinator, the bumblebee Bombus terrestris L.

The primitively social bumblebee Bombus terrestris is an ecological model species as well as an important agricultural pollinator. As part of the ongoing development of genomic resources for this model organism, we have constructed a publicly available bacterial artificial chromosome (BAC) library from males of a field-derived colony. We have shown that this library has a high coverage, which allows any particular sequence to be retrieved from at least one clone with a probability of 99.7%. We have further demonstrated the library’s usefulness by successfully screening it with probes derived both from previously described B. terrestris genes and candidate genes from another bumblebee species and the honeybee. This library will facilitate genomic studies in B. terrestris and will allow for novel comparative studies in the social Hymenoptera. Received 12 March 2008; revised 13 September 2008; accepted 17 September 2008. R. Schmid-Hempel: contact for data repository of this BAC library