Phenotypic variation in male and worker encapsulation response in the bumblebee Bombus terrestris

Abstract.  1. Life-history traits such as immunity are often characterised by the presence of large phenotypic variation, but it often remains unclear how and why this variation is maintained by selection.
2. Here an annual social insect, the bumblebee Bombus terrestris , was used to study variation in encapsulation response of males and workers. Bumblebees are a suitable system to study offspring immunity because they are host to a broad variety of different parasites. Bumblebee males, in particular, have a long lifespan compared with other social insect males and their immunity should therefore be an important element for colony reproductive success.
3. Encapsulation response, which was used here as a measurement for the generalised immune defence capacity of an individual, was found to be a highly variable trait. High levels of worker response correlated with low levels of colony parasitism rates.
4. Encapsulation response was found to be (a) lower in males compared with sister workers, and (b) lower in late-produced cohorts compared with early ones.
5. In colonies with delayed sexual reproduction, males had a lower encapsulation response. Thus, investments into immunity seemed reduced in later male cohorts and those eclosing later in the season, perhaps because males had a shorter expected remaining time to acquire matings. The results presented add further evidence that immune defence is a key variable defining colony fitness in social insects.     

中国明亮熊蜂复合种的分子鉴定及分布特性

【目的】为了探明中国境内明亮熊蜂复合种Bombus lucorum complex的物种组成及其分布特性。【方法】利用DNA条形码技术对来源于中国境内的明亮熊蜂复合种的223个标本进行了鉴定, 并根据鉴定结果分析了各物种的分布特性。【结果】中国境内明亮熊蜂复合种包括地熊蜂B. terrestris、 隐熊蜂B. cryptarum、 明亮熊蜂B. lucorum和长翅熊蜂B. longipennis。4种熊蜂共获得160条DNA条形码序列, 包含36个单倍型, 其中B. longipennis单倍型最多, 为14个; B. terrestris最少, 为3个。B. terrestris和B. lucorum亲缘关系最远, 其遗传距离为0.077; B. lucorum和B. longipennis亲缘关系最近, 其遗传距离为0.034。系统进化分析（贝叶斯法和最大似然法）表明, 4种熊蜂的分化先后顺序为B. terrestris, B. longipennis, B. lucorum和B. cryptarum。4种熊蜂的分布格局不同, 平均分布海拔高度由高到低的顺序依次为B. longipennis, B. cryptarum, B. lucorum和B. terrestris。B. longipennis发现于高海拔的青藏高原东部及南部地区, 而B. terrestris仅发现于低海拔的新疆西北部边界地带。【结论】本研究探明了中国境内明亮熊蜂复合种的分类地位及其分布特性, 为深入研究该类群的资源保护和农业授粉利用奠定了基础。     

Colony performance and immunocompetence of a social insect, Bombus terrestris, in poor and variable environments

1. Feeding and defence against parasites are among the major simultaneous requirements for survival and successful reproduction of any animal, including colonies of social insects. Harsh environments may therefore not only negatively affect overall success, but force animals to allocate resources in different ways, for example, to reduce immune defence under adverse conditions while maintaining growth rate.
2. This study examines how colony growth, reproduction and immune defence of the bumble bee, Bombus terrestris , varies with environmental condition. In particular, the manner in which limited or unpredictable food availability affects the number and size of workers and sexuals produced, the timing of reproduction, and the level of encapsulation of a novel antigen was tested. A new experimental paradigm was applied by splitting colonies into two halves while ensuring integrity by regular swaps of the residing queen.
3. The results show that limited food availability, as expected, led to fewer and smaller workers and sexuals. Unpredictable food availability, in contrast, was associated with higher numbers of workers and heavier sexuals. Also, reproductive success was highest under unpredictable conditions, due to an increased rate of collection of resources.
4. The timing of reproduction did not vary among treatments. Contrary to expectation, also immunocompetence did not vary among treatments.     

Size-related selection of food plants by bumblebees

Abstract. 1. A positive correlation between the tongue length of conspecific workers collecting nectar from seven plant species and the corolla length of the flowers probed was found for B.lapidarius and B.pascuomm but not B.terrestris . No simple relationship was found between the volume, sugar weight or concentration of nectar in flowers and the tongue or wing length of probing bees.
2. B.terrestris workers collecting pollen from four plant species producing pollen only, were found to differ in size according to the type of pollen presentation mechanism and the pollen content per flower. Body size variation was also related to the foraging of pollen plus nectar from two other plant species.     

Genetic analysis of spatial foraging patterns and resource sharing in bumble bee pollinators

Conservation biologists, evolutionary ecologists and agricultural biologists require an improved understanding of how pollinators utilize space and share resources. Using microsatellite markers, we conducted a genetic analysis of space use and resource sharing at several spatial scales among workers of two ecologically dissimilar bumble bee species (Bombus terrestris and B. pascuorum) foraging in an urban landscape (London, UK). At fine scales, the relatedness of workers visiting small patches of flowers did not differ significantly from zero. Therefore, colonies shared flower patches randomly with other colonies, suggesting that worker scent-marks deterring visits to unrewarding flowers have not evolved as signals benefiting nestmates. To investigate space use at intermediate scales, we developed a program based on Thomas & Hill's maximum likelihood sibship reconstruction method to estimate the number of colonies utilizing single sites. The average number of colonies (95% confidence limits) sending workers to forage at sites of approximately 1 ha in area was 96 colonies (84-118) in B. terrestris and 66 colonies (61-76) in B. pascuorum. These values are surprisingly high and suggested that workers travelled far from their colonies to visit the sites. At the landscape scale, there was little or no genetic differentiation between sites. We conclude that urban habitats support large bumble bee populations and are potentially valuable in terms of bumble bee conservation. In addition, bumble bee-mediated gene flow in plants is likely to occur over large distances and plant-bumble bee conservation requires landscape-scale action.     

In vivo dynamics of an immune response in the bumble bee Bombus terrestris

Concepts from evolutionary ecology have recently been applied to questions of immune defences. However, an important but often neglected aspect is the temporal dynamics of the simple immune measures used in ecological studies. Here, we present observations for workers of the bumble bee Bombus terrestris on the dynamics of the phenoloxidase (PO) system, antibacterial activity, and the total number of haemocytes following a challenge with immune elicitors (LPS, Laminarin), over a time-span ranging from 1min to 14 days. The dynamics of the PO measurement showed a complex pattern and was correlated with haemocyte counts. Antibacterial activity, on the other hand, increased sharply between 2 and 24h post-challenge followed by a slow decrease. Surprisingly, the effects of a challenge lasted up to 14 days.     

Monoandry and polyandry in bumble bees (Hymenoptera; Bombinae) as evidenced by highly variable microsatellites

Highly variable microsatellites enabled a precise assessment of the number of queen matings in the colonies of five bumble bee species. Fifteen of the sixteen microsatellites initially cloned from B. terrestris had flanking regions similar enough to allow PCR amplification on the other Bombus species analysed. The microsatellites selected for intracolony study (four per species) were characterized by a high heterozygosity (0.58–0.93) and a large number of alleles (3–18) in the local populations from which the colonies originated. A single male appeared to have inseminated the queens in the colonies of four species, B. terrestris, B. lucorum, B. lapidarius and B. pratorum, which belong to three subgenera, whereas two of the three analysed colonies of B. hypnorum were polyandrous (minimum number of two and four patrilines, respectively).     

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.     

Foraging trip duration of bumblebees in relation to landscape-wide resource availability

Abstract.  1. The study tested the hypotheses that bumblebees have shorter foraging trips in environments that provide abundant resources than in environments that provide sparse resources, and that shorter foraging trips translate into greater colony growth.
2. Six even-aged Bombus terrestris colonies were established in contrasting resource environments. Three colonies had access to abundant resources ( Phacelia tanacetifolia fields with high flower densities), and three colonies were placed in an environment with sparse resources (scattered semi-natural habitats with food plants at lower densities).
3. A total of 870 foraging trips of 220 marked B. terrestris foragers were observed using automated camcorder recordings.
4. The duration of foraging trips was shorter in environments with abundant resources (66 ± 4.6 min) than in environments with sparse resources (82 ± 3.7 min). Within 34 days colonies that had access to abundant resources gained significantly more weight (129 ± 40 g) than colonies foraging on sparse resources (19 ± 7 g).
5. Thus, the spatial distribution and quality of resources at landscape level affected the duration of foraging trips and the colony growth. It was concluded that future conservation schemes need to improve the spatial and temporal availability of resources in agricultural landscapes to counteract the ongoing decline of bumblebees.     

Juvenile hormone titers,juvenile hormone biosynthesis,ovarian development and social environment in Bombus terrestris

The effects of the social environment and age on juvenile hormone (JH) and reproduction were investigated by measuring ovarian development, hemolymph levels of JH III, and rates of JH biosynthesis from the same individual bumble bees (Bombus terrestris). Differences in social environment were associated with differences in rates of JH biosynthesis, JH titer and ovarian development. Young queenless workers had a higher rate of JH biosynthesis, JH titer and ovarian development than queenright (QR) workers of similar age. Dominant workers in QR colonies had a higher rate of JH biosynthesis, JH titer and ovarian development than low ranked workers of similar size. There was a positive correlation between JH titer and ovarian development, but no correlation between rate of JH biosynthesis and ovarian development or between JH biosynthesis and JH titer. Both JH titer and rate of JH biosynthesis increased with age from emergence to 3 days of age, but 6-day-old workers, egg-laying workers, and actively reproducing queens had high JH titers and highly developed ovaries but low rates of JH biosynthesis. These results show that reproduction in B. terrestris is strongly affected by the social environment and the influence of the environment on reproduction is mediated by JH. Our data also indicate that the rate of JH biosynthesis measured in vitro is not a reliable indicator of JH titer or ovarian development in B. terrestris; possible reasons are discussed.     

Differential virulence in a multiple‐host parasite of bumble bees: resolving the paradox of parasite survival?

Parasite virulence determines both the impact that parasites have on their hosts and parasite fitness. While most studies of virulence have involved single-species host–parasite interactions, the majority of parasites are likely to use multiple concurrent host species. Our understanding of how this impacts on parasite epidemiology and virulence is limited. Using the bumble bee Bombus lucorum , which exists in sympatry with B. terrestris in multi-species assemblages, and their generalist micosporidian parasite Nosema bombi , we tested whether the apparent paradox of parasite maintenance due to parasite virulence in a single host, B. terrestris , could be resolved through understanding the parasite's virulence in this sympatric host species. Nosema bombi significantly impacted colony growth, individual longevity, and individual development in B. lucorum . However, these effects were different both qualitatively and quantitatively to the parasite's impact in B. terrestris . Infected colonies of B. lucorum successfully produced both male and female reproductives, and infected female reproductives were capable of successful mating. Variation in life-history across host species may explain differences in the virulence, or impact of the parasite in B. terrestris and B. lucorum , with species with shorter life-cycles being more likely to transfer the parasite from one annual generation to the next. These results suggest that to understand the virulence and epidemiology of multi-host parasites we need to examine their ecological interactions across their various host species.     

Bumblebee commercialization will cause worldwide migration of parasitic mites

We investigated natural populations of three Japanese native bumblebee species to determine the status of infestation by a tracheal mite, Locustacarus buchneri, which we had earlier detected in introduced commercial colonies of the European bumblebee, Bombus terrestris. We also investigated mite infestation in commercial colonies of a Japanese native species, B. ignitus, which are mass-produced in the Netherlands and reimported into Japan. We detected the mite in both natural and commercial colonies of the Japanese species. Comparison of 555 bp sequences of the mitochondrial DNA (mtDNA) cytochrome oxidase 1 (CO1) gene from the mite showed that there were seven haplotypes, on the basis of combinations of substitutions at eight sites in the gene. The haplotypes of the mites in the Japanese native bumblebees and the haplotypes of the mites in B. terrestris did not overlap; however, mtDNA of mites detected in the commercial colonies of B. ignitus possessed the same sequence as a European haplotype. These results indicate that transportation of bumblebee colonies will cause overseas migration of parasitic mites of different origins.     

The effects of age and social interactions on innate immunity in a leaf-cutting ant

Both developmental and environmental factors shape investment in costly immune defences. Social insect workers have different selection pressures on their innate immune system compared to non-social insects because workers do not reproduce and their longevity affects the fitness of relatives. Furthermore, hygienic behavioural defences found in social insects can result in increased survival after fungal infection, although it is not known if there is modulation in physiological immune defence associated with group living vs. solitary living.Here we investigated whether physiological immune defence is affected by both age and the short-term presence or absence of nestmates in the leaf-cutting ant Acromyrmex octospinosus. We predicted that older ants would show immune senescence and that group living may result in prophylactic differences in immune defence compared to solitarily kept ants. We kept old and young workers alone or in nestmate groups for 48 h and assayed a key innate immune system enzyme, expressing phenoloxidase (PO) and its stored precursor (proPO), a defence that acts immediately, i.e. it is constitutive. Short-term solitary living did not affect PO or proPO levels relative to group living controls and we found no evidence for immunosenescence in proPO. However, we found a significant increase in active PO in older workers, which is consistent with two non-mutually exclusive explanations: it could be an adaptive response or indicative of immunosenescence. Our results suggest that future studies of immunosenescence should consider both active immune effectors in the body, such as PO, and the stored potential to express immune defences, such as proPO.     

明亮熊蜂繁育室内印度谷斑螟的形态特征与生物学特性

明亮熊蜂Bombus lucorum L.是一种重要的温室果菜传粉昆虫,印度谷斑螟Plodia interpunctella(Hbner)是危害明亮熊蜂繁育的主要害虫之一。2年的研究结果表明,印度谷斑螟的形态特征与危害蜜蜂巢房的大蜡螟Gallerie mellonellaL.和小蜡螟Achroia grisella Fabricius明显不同。印度谷斑螟以饲喂熊蜂的花粉为载体传播进熊蜂繁育室,在熊蜂繁育室内可以连续繁殖,1年发生6～8代,且世代重叠。印度谷斑螟幼虫主要以蜂群内的剩花粉为食,当花粉不足时,就开始取食巢房和蜂蛹,对熊蜂的规模化繁育危害较大,每年的5～8月、11～2月为危害高峰期。印度谷斑螟的发育受环境和食物的影响很大,在温度为28℃、相对湿度为60%熊蜂繁育室内,印度谷斑螟的卵期为4d,幼虫期为19～23d,蛹期为11d,成虫期为4～20d。成虫羽化后3～4h即可进行交配,雌蛾交配后当天就开始产卵,产卵期4～10d不等,平均产卵量107.8粒。在熊蜂繁育室消毒期间,食物短缺,印度谷斑螟幼虫进入休眠状态,各龄幼虫均可发生休眠现象。印度谷斑螟不危害蜜蜂,在蜜蜂巢房中不能存活。     

Immune responses of bumblebee workers as a function of individual and colony age: senescence versus plastic adjustment of the immune function

Senescence may evolve in response to externally imposed schedules of survival and reproduction. It may result from cumulative damage to several tissues and organs leading to the progressive malfunction of somatic defences including the immune system, which is a relevant and accessible trait to study the evolution and the mechanisms of senescence.
In social insects, reproduction of workers is negligible and their fitness relies on the colony reproductive success, which depends on workers' longevity. Consequently, evolution of senescence in workers will rely on age-dependent survival rate and the residual reproductive value of the colony. Using the bumblebee, Bombus terrestris , as model system, we investigated changes in constitutive and inducible immune defences. Such changes were monitored in relation to worker age and colony age and as well under a high, persistent level of parasitism. The results show that the inducible production of antibacterial peptides was not affected by the age of individual worker or the age of their colony. In contrast, constitutive defences such as haemocyte density and phenoloxidase (PO) activity decline with individual worker age and potential accumulation of physiological defects due to high and persistent levels of parasitism did not explain this pattern of immunosenescence. Interestingly, levels of worker constitutive defences were found to increase with colony age. We found antibacterial activity was strongly traded-off against PO activity. Such a relationship was not caused by resource shortage. Overall, these data suggest that patterns of variation of immune defences with worker age and colony age are likely the result of plastic temporal adjustments of immune responses in accordance with life history predictions. Hence, control of pathogenic threat in a social system may involve optimal adjustments of immune defences at both the individual and colony levels.     

授粉用明亮熊蜂的人工饲养技术

为提高授粉用熊蜂人工繁育效率,降低生产成本,对明亮熊蜂Bombus lucorumL.人工饲养中诱导蜂王产卵和人工控制下的交配2个技术环节进行了研究。结果表明,采用诱导器和伴饲2~3只蜜蜂工蜂的诱导方法,蜂王产卵率和蜂群成群率最高,成群时间最短;使用塑料诱导器和木制诱导器的效果没有显著差异。将8~9日龄的蜂王和11~12日龄的雄蜂放入交配笼时,其交配成功率最高。交配笼内的蜂王数量应控制在30只/m3左右。在晴天,交配笼放置在室外(758 000 lux),其交配成功率最高;在阴天,交配笼应放在交配室内,并开启荧光灯照明(35 000 lux),其交配成功率最高。     

11种常用农药对地熊蜂工蜂的毒性和风险评估

【目的】评估常用农药对地熊蜂Bombus terrestris的生态风险,为设施大棚合理施用农药提供科学依据。【方法】分别采用饲喂法和接触法测定了6种杀虫剂(虫螨腈、高效氯氟氰菊酯、氟吡呋喃酮、螺虫乙酯、异丙威和除虫脲)、3种杀螨剂(丁氟螨酯、唑螨酯和联苯肼酯)及2种杀菌剂(春雷霉素和啶酰菌胺)共11种常用农药对地熊蜂成年工蜂的急性经口和急性接触毒性,并评估其生态风险性。【结果】11种农药经饲喂法测定,对地熊蜂工蜂的急性经口毒性除高效氯氟氰菊酯、异丙威和虫螨腈为高毒,氟吡呋喃酮和唑螨酯为中毒外,其余药物均为低毒。经接触法测定,对地熊工蜂的急性接触毒性除高效氯氟氰菊酯和异丙威为高毒,虫螨腈为中毒外,其余药物均为低毒。生态风险评估表明,对地熊蜂工蜂而言,异丙威和高效氯氟氰菊酯的经口与接触毒性为中风险,氟吡呋喃酮、啶酰菌胺、除虫脲、唑螨酯、联苯肼酯、螺虫乙酯、春雷霉素、丁氟螨酯的经口与接触毒性为低风险;虫螨腈的经口毒性为中风险,接触毒性为低风险。【结论】在设施作物花期使用地熊蜂授粉时,建议禁用异丙威、高效氯氟氰菊酯和虫螨腈这3种存在中风险的农药,慎重使用氟吡呋喃酮和唑螨酯这2种农药,以避免对地熊蜂造成危害,而另外6种低毒农药可根据田间情况合理施用,并可采取通风晾晒、设置间隔期等方式降低农药对地熊蜂的生态风险。     

Kin-selected conflict in the bumble-bee Bombus terrestris (Hymenoptera: Apidae)

Kin selection theory predicts conflict in social Hymenoptera between the queen and workers over male parentage because each party is more closely related to its own male offspring. Some aspects of the reproductive biology of the bumble-bee Bombus terrestris support kin selection theory but others arguably do not. We present a novel hypothesis for how conflict over male parentage should unfold in B. terrestris colonies. We propose that workers delay laying eggs until they possess information showing that egg laying suits their kin-selected interests. In colonies where queens start to lay haploid eggs early, we hypothesize that this occurs when workers detect the presence of queen-produced male brood in the brood's larval stage. In colonies where queens start to lay haploid eggs late, we hypothesize that it occurs when workers detect a signal from the queen to female larvae to commence development as queens. Our hypothesis accounts for previously unexplained aspects of the timing of reproductive events in B. terrestris, provides ultimate explanations for the results of a recent study of mechanisms underlying queen-worker conflict and helps explain this species' characteristic bimodal (split) sex ratios. Therefore, kin selection theory potentially provides a good explanation for reproductive patterns in B. terrestris.     

欧洲熊蜂Bombus terretris入侵研究进展

欧洲熊蜂Bombus terrestris被出口到自然分布区外为作物传粉,现已在澳大利亚、新西兰、日本、以色列、智利和阿根廷等国家建立野生种群.欧洲熊蜂与本土传粉生物竞争蜜粉源植物,传播病虫害,与本土熊蜂杂交产生不育子代.欧洲熊蜂入侵可能引起本地传粉昆虫牛物多样性降低,危害生态系统传粉功能的安全.中国从1998年开始引入欧洲熊蜂为大棚作物传粉,但日前尚无欧洲熊蜂入侵我国的报道.中国是熊蜂生物多样性最丰富的国家之一,具有欧洲熊蜂适生环境,欧洲熊蜂有可能入侵我国.防止欧洲熊蜂入侵最重要和最有效的办法是减少进口该物种和进行科学的放蜂管理.     

Ecdysteroid titer, ovary status, and dominance in adult worker and queen bumble bees (Bombus terrestris)

The role of ecdysteroids in the regulation of dominance and reproduction in social Hymenoptera is little explored. In the current study we compared ecdysteroid titers in hemolymph of individual queen and worker bumble bees (Bombus terrestris) that differ in their behavior, reproductive status and social environment. Egg-laying queens that head colonies and have ovaries exhibiting all stages of follicle development, had a higher ecdysteroid titer than virgin queens whose ovaries contained only follicles at initial stages. In workers, the relationship between ecdysteroid titers and follicle development appears to be more complex and to be influenced by the bee's social status and social environment. Shortly after emergence, young workers had only follicles at the initial stages of oogenesis and they exhibited a low ecdysteroid titer. No significant correlation was detected between ovary status and ecdysteroid titer in workers, with some workers showing activated ovaries but low ecdysteroid titers. However, at six days of age, a trend towards higher ecdysteroid titer was observed for workers in queenless groups, a condition characterized by rapid follicle development relative to queenright conditions. In these queenless groups, high social status was associated with high ecdysteroid titers. By contrast, in queenright workers ecdysteroid titers were low, even for bees with presumably high social status that had activated ovaries and were observed performing oviposition behavior. This study suggests that ecdysteroids are involved in regulation of reproduction in B. terrestris.