Interspecific utilisation of wax in comb building by honeybees

Beeswaxes of honeybee species share some homologous neutral lipids; but species-specific differences remain. We analysed behavioural variation for wax choice in honeybees, calculated the Euclidean distances for different beeswaxes and assessed the relationship of Euclidean distances to wax choice. We tested the beeswaxes of Apis mellifera capensis, Apis florea, Apis cerana and Apis dorsata and the plant and mineral waxes Japan, candelilla, bayberry and ozokerite as sheets placed in colonies of A. m. capensis, A. florea and A. cerana. A. m. capensis accepted the four beeswaxes but removed Japan and bayberry wax and ignored candelilla and ozokerite. A. cerana colonies accepted the wax of A. cerana, A. florea and A. dorsata but rejected or ignored that of A. m. capensis, the plant and mineral waxes. A. florea colonies accepted A. cerana, A. dorsata and A. florea wax but rejected that of A. m. capensis. The Euclidean distances for the beeswaxes are consistent with currently prevailing phylogenies for Apis. Despite post-speciation chemical differences in the beeswaxes, they remain largely acceptable interspecifically while the plant and mineral waxes are not chemically close enough to beeswax for their acceptance.     

A bee in the corridor: centering and wall-following

In an attempt to better understand the mechanism underlying lateral collision avoidance in flying insects, we trained honeybees (Apis mellifera) to fly through a large (95-cm wide) flight tunnel. We found that, depending on the entrance and feeder positions, honeybees would either center along the corridor midline or fly along one wall. Bees kept following one wall even when a major (150-cm long) part of the opposite wall was removed. These findings cannot be accounted for by the “optic flow balance” hypothesis that has been put forward to explain the typical bees’ “centering response” observed in narrower corridors. Both centering and wall-following behaviors are well accounted for, however, by a control scheme called the lateral optic flow regulator, i.e., a feedback system that strives to maintain the unilateral optic flow constant. The power of this control scheme is that it would allow the bee to guide itself visually in a corridor without having to measure its speed or distance from the walls.     

Heat-balling wasps by honeybees

Defensiveness of honeybee colonies of Apis cerana and Apis mellifera (actively balling the wasps but reduction of foraging) against predatory wasps, Vespa velutina, and false wasps was assessed. There were significantly more worker bees in balls of the former than latter. Core temperatures in a ball around a live wasp of A. cerana were significantly higher than those of A. mellifera, and also significantly more when exposed to false wasps. Core temperatures of bee balls exposed to false wasps were significantly lower than those exposed to V. velutina for both A. cerana and for A. mellifera. The lethal thermal limits for V. velutina, A. cerana and A. mellifera were significantly different, so that both species of honeybees have a thermal safety factor in heat-killing such wasp predators. During wasps attacks at the hives measured at 3, 6 and 12 min, the numbers of Apis cerana cerana and Apis cerana indica bees continuing to forage were significantly reduced with increased wasp attack time. Tropical lowland A. c. indica reduced foraging rates significantly more than the highland A. c. cerana bees; but, there was no significant effect on foraging by A. mellifera. The latency to recovery of honeybee foraging was significantly greater the longer the duration of wasp attacks. The results show remarkable thermal fine-tuning in a co-evolving predator–prey relationship.     

Heat and carbon dioxide generated by honeybees jointly act to kill hornets

We have found that giant hornets (Vespa mandarinia japonica) are killed in less than 10 min when they are trapped in a bee ball created by the Japanese honeybees Apis cerana japonica, but their death cannot be solely accounted for by the elevated temperature in the bee ball. In controlled experiments, hornets can survive for 10 min at the temperature up to 47°C, whereas the temperature inside the bee balls does not rise higher than 45.9°C. We have found here that the CO₂ concentration inside the bee ball also reaches a maximum (3.6 ± 0.2%) in the initial 0–5 min phase after bee ball formation. The lethal temperature of the hornet (45–46°C) under conditions of CO₂ concentration (3.7 ± 0.44%) produced using human expiratory air is almost the same as that in the bee ball. The lethal temperature of the honeybee is 50–51°C under the same air conditions. We concluded that CO₂ produced inside the bee ball by honeybees is a major factor together with the temperature involved in defense against giant hornets.     

Bee-hawking by the wasp, <Emphasis Type="BoldItalic">Vespa velutina</Emphasis>, on the honeybees <Emphasis Type="BoldItalic">Apis cerana</Emphasis> and <Emphasis Type="BoldItalic">A</Emphasis>. <Emphasis Type="BoldItalic">mellifera</Emphasis>

The vespine wasps, Vespa velutina, specialise in hawking honeybee foragers returning to their nests. We studied their behaviour in China using native Apis cerana and introduced A. mellifera colonies. When the wasps are hawking, A. cerana recruits threefold more guard bees to stave off predation than A. mellifera. The former also utilises wing shimmering as a visual pattern disruption mechanism, which is not shown by A. mellifera. A. cerana foragers halve the time of normal flight needed to dart into the nest entrance, while A. mellifera actually slows down in sashaying flight manoeuvres. V. velutina preferentially hawks A. mellifera foragers when both A. mellifera and A. cerana occur in the same apiary. The pace of wasp-hawking was highest in mid-summer but the frequency of hawking wasps was three times higher at A. mellifera colonies than at the A. cerana colonies. The wasps were taking A. mellifera foragers at a frequency eightfold greater than A. cerana foragers. The final hawking success rates of the wasps were about three times higher for A. mellifera foragers than for A. cerana. The relative success of native A. cerana over European A. mellifera in thwarting predation by the wasp V. velutina is interpreted as the result of co-evolution between the Asian wasp and honeybee, respectively.     

Indirect interactions between invasive and native plants via pollinators

In generalised pollination systems, the presence of alien plant species may change the foraging behaviour of pollinators on native plant species, which could result in reduced reproductive success of native plant species. We tested this idea of indirect interactions on a small spatial and temporal scale in a field study in Mauritius, where the invasive strawberry guava, Psidium cattleianum, provides additional floral resources for insect pollinators. We predicted that the presence of flowering guava would indirectly and negatively affect the reproductive success of the endemic plant Bertiera zaluzania, which has similar flowers, by diverting shared pollinators. We removed P. cattleianum flowers within a 5-m radius from around half the B. zaluzania target plants (treatment) and left P. cattleianum flowers intact around the other half (control). By far, the most abundant and shared pollinator was the introduced honey bee, Apis mellifera, but its visitation rates to treatment and control plants were similar. Likewise, fruit and seed set and fruit size and weight of B. zaluzania were not influenced by the presence of P. cattleianum flowers. Although other studies have shown small-scale effects of alien plant species on neighbouring natives, we found no evidence for such negative indirect interactions in our system. The dominance of introduced, established A. mellifera indicates their replacement of native insect flower visitors and their function as pollinators of native plant species. However, the pollination effectiveness of A. mellifera in comparison to native pollinators is unknown. Christine B. Müller, deceased 7 March 2008.     

Plant origin of Okinawan propolis: honeybee behavior observation and phytochemical analysis

Propolis is a natural resinous product collected by honeybees from certain plants. It has gained popularity as a food and alternative medicine. Poplar and Baccharis are well known as the source plants of European and Brazilian propolis, respectively. However, the propolis from Okinawa, Japan, contains some prenylflavonoids not seen in other regions such as Europe and Brazil, suggesting that the plant origin of Okinawan propolis is a particular plant that grows in Okinawa. To identify the plant origin of Okinawan propolis, we observed the behavior of honeybees as they collected material from plants and caulked it inside the hive. Honeybees scraped resinous material from the surface of plant fruits of Macaranga tanarius and brought it back to their hive to use it as propolis. We collected samples of the plant and propolis, and compared their constituents by high-performance liquid chromatography with a photo-diode array detector. We also compared their 1,1-diphenyl-2-picryl-hydrazyl radical scavenging activity. The chemical constituents and biological activity of the ethanol extracts of the plant did not differ from those of propolis. This indicates directly that the plant origin of Okinawan propolis is M. tanarius. S. K. and J. N. contributed equally to this work.     

Temporal genetic structure of a drone congregation area of the giant Asian honeybee (Apis dorsata)

The giant Asian honeybee (Apis dorsata), like all other members of the genus Apis, has a complex mating system in which the queens and males (drones) mate at spatially defined drone congregation areas (DCAs). Here, we studied the temporal genetic structure of a DCA of A. dorsata over an 8-day time window by the genotyping of sampled drones with microsatellite markers. Analysis of the genotypic data revealed a significant genetic differentiation between 3 sampling days and indicated that the DCA was used by at least two subpopulations at all days in varying proportions. The estimation of the number of colonies which used the DCA ranged between 20 and 40 colonies per subpopulation, depending on the estimation procedure and population. The overall effective population size was estimated as high as N _e=140. The DCA seems to counteract known tendencies of A. dorsata for inbreeding within colony aggregations by facilitating gene flow among subpopulations and increasing the effective population size.     

Africanized honeybees are slower learners than their European counterparts

Does cognitive ability always correlate with a positive fitness consequence? Previous research in both vertebrates and invertebrates provides mixed results. Here, we compare the learning and memory abilities of Africanized honeybees (Apis mellifera scutellata hybrid) and European honeybees (Apis mellifera ligustica). The range of the Africanized honeybee continues to expand, superseding the European honeybee, which led us to hypothesize that they might possess greater cognitive capabilities as revealed by a classical conditioning assay. Surprisingly, we found that fewer Africanized honeybees learn to associate an odor with a reward. Additionally, fewer Africanized honeybees remembered the association a day later. While Africanized honeybees are replacing European honeybees, our results show that they do so despite displaying a relatively poorer performance on an associative learning paradigm.     

Behavioral genomics of honeybee foraging and nest defense

The honeybee has been the most important insect species for study of social behavior. The recently released draft genomic sequence for the bee will accelerate honeybee behavioral genetics. Although we lack sufficient tools to manipulate this genome easily, quantitative trait loci (QTLs) that influence natural variation in behavior have been identified and tested for their effects on correlated behavioral traits. We review what is known about the genetics and physiology of two behavioral traits in honeybees, foraging specialization (pollen versus nectar), and defensive behavior, and present evidence that map-based cloning of genes is more feasible in the bee than in other metazoans. We also present bioinformatic analyses of candidate genes within QTL confidence intervals (CIs). The high recombination rate of the bee made it possible to narrow the search to regions containing only 17–61 predicted peptides for each QTL, although CIs covered large genetic distances. Knowledge of correlated behavioral traits, comparative bioinformatics, and expression assays facilitated evaluation of candidate genes. An overrepresentation of genes involved in ovarian development and insulin-like signaling components within pollen foraging QTL regions suggests that an ancestral reproductive gene network was co-opted during the evolution of foraging specialization. The major QTL influencing defensive/aggressive behavior contains orthologs of genes involved in central nervous system activity and neurogenesis. Candidates at the other two defensive-behavior QTLs include modulators of sensory signaling (Am5HT ₇ serotonin receptor, AmArr4 arrestin, and GABA-B-R1 receptor). These studies are the first step in linking natural variation in honeybee social behavior to the identification of underlying genes.     

RNAi-mediated silencing of vitellogenin gene function turns honeybee (Apis mellifera) workers into extremely precocious foragers

The switch from within-hive activities to foraging behavior is a major transition in the life cycle of a honeybee (Apis mellifera) worker. A prominent regulatory role in this switch has long been attributed to juvenile hormone (JH), but recent evidence also points to the yolk precursor protein vitellogenin as a major player in behavioral development. In the present study, we injected vitellogenin double-stranded RNA (dsVg) into newly emerged worker bees of Africanized genetic origin and introduced them together with controls into observation hives to record flight behavior. RNA interference-mediated silencing of vitellogenin gene function shifted the onset of long-duration flights (>10 min) to earlier in life (by 3–4 days) when compared with sham and untreated control bees. In fact, dsVg bees were observed conducting such flights extremely precociously, when only 3 days old. Short-duration flights (<10 min), which bees usually perform for orientation and cleaning, were not affected. Additionally, we found that the JH titer in dsVg bees collected after 7 days was not significantly different from the controls. The finding that depletion of the vitellogenin titer can drive young bees to become extremely precocious foragers could imply that vitellogenin is the primary switch signal. At this young age, downregulation of vitellogenin gene activity apparently had little effect on the JH titer. As this unexpected finding stands in contrast with previous results on the vitellogenin/JH interaction at a later age, when bees normally become foragers, we propose a three-step sequence in the constellation of physiological parameters underlying behavioral development. David Santos Marco Antonio and Karina Rosa Guidugli-Lazzarini contributed equally to the present study.     

A male gift to its partner? Cyanogenic glycosides in the spermatophore of longwing butterflies (<Emphasis Type="Italic">Heliconius</Emphasis>)

Males of several insect species transfer nuptial gifts to females during mating, typically in the form of a protein-rich spermatophore. In chemically defended species, males could potentially enhance such a gift with chemicals that help protect the female, her eggs, or both. This was shown for lepidopteran species that accumulate pyrrolizidine alkaloids. Most Heliconius butterflies are presumably protected from predators by virtue of de novo synthesized and/or sequestered cyanogenic glycosides. Males of Heliconius species are known to transfer nutritional gifts to the females but whether defensive chemicals could also be transferred is not known. To ascertain whether transfer of cyanogens occurs, we dissected freshly mated females from nine different Heliconius species and analyzed spermatophores for cyanogenic glycosides. We found cyanogens in the spermatophores of all nine species. This is the first time cyanogenic glycosides are reported in the spermatophores of arthropods. We discuss the implications of these findings for Heliconius biology and for other cyanogenic insects as well. We suggest that chemically defended species commonly lace their nuptial gifts with defensive chemicals to improve gift quality.     

Non-specific association between filamentous bacteria and fungus-growing ants

Fungus-growing ants and their fungal cultivar form a highly evolved mutualism that is negatively affected by the specialized parasitic fungus Escovopsis. Filamentous Pseudonocardia bacteria occurring on the cuticle of attine ants have been proposed to form a mutualistic interaction with these ants in which they are vertically transmitted (i.e. from parent to offspring colonies). Given a strictly vertical transmission of Pseudonocardia, the evolutionary theory predicts a reduced genetic variability of symbionts among ant lineages. The aim of this study was to verify whether actinomycetes, which occur on Acromyrmex octospinosus leaf-cutting ants, meet this expectation by comparing their genotypic variability with restriction fragment length polymorphisms. Multiple actinomycete strains could be isolated from both individual ant workers and colonies (one to seven strains per colony). The colony specificity of actinomycete communities was high: Only 15% of all strains were isolated from more than one colony, and just 5% were present in both populations investigated. Partial sequencing of 16S ribosomal deoxyribonucleic acid of two of the isolated strains assigned both of them to the genus Streptomyces. Actinomycetes could also be isolated from workers of the two non-attine ant species Myrmica rugulosa and Lasius flavus. Sixty-two percent of the strains derived from attine ants and 80% of the strains isolated from non-attine ants inhibited the growth of Escovopsis. Our data suggest that the association between attine ants and their actinomycete symbionts is less specific then previously thought. Soil-dwelling actinomycetes may have been dynamically recruited from the environment (horizontal transmission), probably reflecting an adaptation to a diverse community of microbial pathogens.     

Oviposition by small hive beetles elicits hygienic responses from Cape honeybees

Two novel behaviours, both adaptations of small hive beetles (Aethina tumida Murray) and Cape honeybees (Apis mellifera capensis Esch.), are described. Beetles puncture the sides of empty cells and oviposit under the pupae in adjoining cells. However, bees detect this ruse and remove infested brood (hygienic behaviour), even under such well-disguised conditions. Indeed, bees removed 91% of treatment brood (brood cells with punctured walls caused by beetles) but only 2% of control brood (brood not exposed to beetles). Only 91% of treatment brood actually contained beetle eggs; the data therefore suggest that bees remove only that brood containing beetle eggs and leave uninfected brood alone, even if beetles have accessed (but not oviposited on) the brood. Although this unique oviposition strategy by beetles appears both elusive and adaptive, Cape honeybees are able to detect and remove virtually all of the infested brood.     

Chemical compounds of the foraging recruitment pheromone in bumblebees

When the frenzied and irregular food-recruitment dances of bumblebees were first discovered, it was thought that they might represent an evolutionary prototype to the honeybee waggle dance. It later emerged that the primary function of the bumblebee dance was the distribution of an alerting pheromone. Here, we identify the chemical compounds of the bumblebee recruitment pheromone and their behaviour effects. The presence of two monoterpenes and one sesquiterpene (eucalyptol, ocimene and farnesol) in the nest airspace and in the tergal glands increases strongly during foraging. Of these, eucalyptol has the strongest recruitment effect when a bee nest is experimentally exposed to it. Since honeybees use terpenes for marking food sources rather than recruiting foragers inside the nest, this suggests independent evolutionary roots of food recruitment in these two groups of bees.     

Host pollination mode and mutualist pollinator presence: net effect of internally ovipositing parasite in the fig–wasp mutualism

The Ficus–their specific pollinating fig wasps (Chalcidoidea, Agaonidae) interaction presents a striking example of mutualism. Figs also shelter numerous non-pollinating fig wasps (NPFW) that exploit the fig–pollinator mutualism. Only a few NPFW species can enter figs to oviposit, they do not belong to the pollinating lineage Agaonidae. The internally ovipositing non-agaonid fig wasps can efficiently pollinate the Ficus species that were passively pollinated. However, there is no study to focus on the net effect of these internally ovipositing non-agaonid wasps in actively pollinated Ficus species. By collecting the data of fig wasp community and conducting controlled experiments, our results showed that internally ovipositing Diaziella bizarrea cannot effectively pollinate Ficus glaberrima, an actively pollinated monoecious fig tree. Furthermore, D. bizarrea failed to reproduce if they were introduced into figs without Eupristina sp., the regular pollinator, as all the figs aborted. Furthermore, although D. bizarrea had no effect on seed production in shared figs, it significantly reduced the number of Eupristina sp. progeny emerging from them. Thus, our experimental evidence shows that reproduction in Diaziella depends on the presence of agaonid pollinators, and whether internally ovipositing parasites can act as pollinators depends on the host fig’s pollination mode (active or passive). Overall, this study and others suggest a relatively limited mutualistic role for internally ovipositing fig wasps from non-pollinator (non-Agaonidae) lineages.     

底栖藻对水丝蚓生物扰动效应的抑制研究

霍甫水丝蚓(Limnodrilus hoffmeisteri)是富营养化湖泊中的底栖动物优势种,通过生物扰动可以提高水体营养盐浓度,并促进浮游植物生长.富营养浅水湖泊经生态修复后,水体透明度会得到改善,有利于底栖藻快速生长,从而降低水体营养盐水平.那么,底栖藻能否抑制水丝蚓对富营养水体水质的不良影响?为此,本文开展了双因素(底栖藻和水丝蚓)的室外受控实验,结果表明:在无底栖藻处理中,水丝蚓显著提高了水体总氮(TN)、总磷(TP)、总溶解磷(TDP)和叶绿素a(Chl-a)浓度,同时显著降低水体溶解氧(DO)浓度;而在有底栖藻处理中,水丝蚓对水体TP、DO及Chl-a浓度的影响不显著.研究结果表明,水丝蚓的生物扰动提高了水体营养盐和浮游植物浓度,促进水体富营养化.但底栖藻群落的发展能在一定程度上抑制水丝蚓的生物扰动效应.     

磷差异性调控水稻根际nirK/nirS型反硝化菌组成与丰度

磷作为一种重要的生命元素,对反硝化微生物的活性和功能有重要影响.反硝化功能基因nir K和nir S是编码亚硝酸还原酶的两种同工酶基因,但是,磷对具有同种功能的nir K和nir S型反硝化菌的调控是否存在差异尚不十分清楚.本文采集严重缺磷红壤性水稻土设置水稻盆栽试验,通过外源添加磷肥设置对照(CK,P:0 mg·kg-1),低磷(P1,P:15 mg·kg-1),高磷(P2,P:30 mg·kg-1)这3个磷添加水平,研究不同磷水平对水稻亚硝酸还原酶基因的组成与丰度的调控作用.结果表明,在长期缺磷土壤上施用磷肥对含亚硝酸还原酶基因nir K和nir S的细菌种群的调控作用有明显差异.不管是根际还是非根际土,含nir S的微生物种群均对施磷水平表现敏感,尤其是高磷(P2)水平,施磷可导致nir S丰度提高2～3倍,同时显著改变nir S型反硝化微生物的组成结构.相比之下,施磷后含nir K基因的微生物组成结构和丰度变异较小.与非根际土壤相比,高磷水平条件下根际土中nir S的基因拷贝数和群落结构均发生了显著变化,缺磷和低磷条件下水稻生长只引起根际土nir S种群组成结构发生显著变化,但其丰度与非根际无显著差异.但不同磷水平条件下nir K的基因丰度和组成结构在根际和非根际土之间几乎无显著变化.综上所述,在严重缺磷水稻土中加施磷肥会显著提高水稻根际和非根际土中nir K和nir S型反硝化菌数量,并改变其种群组成结构,且nir S比nir K型种群响应更敏感.不同磷水平条件下的水稻根系生长均显著改变了根际土壤中nir K和nir S种群组成结构,但除了在高磷水平条件下显著增加了nir S丰度外,对nir K和nir S丰度均影响较小.研究结果可为进一步深入探究施肥对土壤反硝化过程的影响提供理论依据.     

Impacts of Climate Change on the Distribution of Larix Spp. and Pinus Sylvestris and Their Climatypes in Siberia

Inter- and intraspecific effects of climate change were assessed for the dominant conifers of Siberia (60–140^∘E and 48–75^∘N): Larix spp. (L. sibirica, L. dahurica, and L. sukaczewii) and Pinus sylvestris . The approach employed a tri-variate (degree-days above 5 ^∘C, degree-days below 0 ^∘C, and a moisture index) estimate of the climatic envelope within which exists the actual ecological distribution of a species and their constituent climatypes (genotypes physiologically attuned to similar environments). Limits of the actual ecological distribution were approximated by reducing the climatic envelope according to effects of permafrost and interspecific competition. Climatypes were mapped within the climatic envelope according to the climatic interval that must separate populations for reasonable assurance of genetic differentiation. This interval was calculated from response functions that related 13-year growth and survival of a species to the difference in climate between the provenance of a climatype and the climate of numerous test sites distributed across Russia. Mapping species' distributions and their climatypes was done for the contemporary climate and for future climates predicted by the HadCM3GGa1 scenario of Hadley Centre. The results showed that if the forests of the future are to reflect the adaptedness of today, the distribution of species will shift and genotypes within species will be redistributed. Some contemporary climatypes are projected to disappear from Siberia while others common elsewhere would evolve. To mitigate these effects, climatypes should be transferred today to the expected future location of their climatic optima, a distance that is likely to approach 700–1200 km for these species.     

Vegetation dynamics after compost amendment in a Mediterranean post-fire ecosystem

On Mediterranean calcareous soils, high fire frequency induces soil impoverishment and the development of stable Quercus coccifera garrigues. Organic amendment could increase soil fertility levels, and could alter the vegetation dynamics and the established dominance relationships. In this study, the plant cover evolution is monitored in an amended burnt shrub during two years. Three treatments are studied: control (D0), 50 t ha⁻¹ (D50) and 100 t ha⁻¹ (D100) of fresh co-composted sewage sludge and greenwastes.First, the spreading process leads to the squashing of the vegetation, whose intensity reaches a threshold on D100 and limits the woody species’ recovery after amendment. Consequently, the dominant herb Brachypodium retusum is favoured. On D50, woody species are favoured compared to herbs, probably due to the space colonization strategy of Q. coccifera after squashing. Thus, compost at both rates favours the two dominant resprouter species until they have colonized all the free space created by squashing.Secondly, compost has some fertilizing effects on Rosmarinus officinalis and Ulex parviflorus, that improves their cover on D50. However, compost also increases U. parviflorus’ sensitivity to drought.At the end of the experiment, a third compost effect appears, as seeder cover becomes greater than D0 on D50. After amendment, compost covers rocky stones, which creates a new territory where species with superficial rooting can establish (mulch effect).