Confinement Behavior of Cape Honey Bees (Apis mellifera capensis Esch.) in Relation to Population Densities of Small Hive Beetles (Aethina tumida Murray)

We quantified the effects of increasing small hive beetle (Aethina tumida Murray) populations on guarding behavior of Cape honey bees (Apis mellifera capensis, an African subspecies). We found more confinement sites (prisons) at the higher (50 beetles per colony) rather than lower (25 beetles per colony) beetle density. The number of beetles per prison did not change with beetle density. There were more guard bees per beetle during evening than morning. Neither guard bee nor beetle behavior varied with beetle density or over time. Forty-six percent of all beetles were found among the combs at the low beetle density and this increased to 58% at the higher one. In neither instance were beetles causing depredation to host colonies. Within the limits of the experiment, guarding behavior of Cape honey bees is relatively unaffected by increasing beetle density (even if significant proportions of beetles reach the combs).     

Behavioral defense strategies of the stingless bee, <Emphasis Type="Italic">Austroplebeia australis</Emphasis>, against the small hive beetle, <Emphasis Type="Italic">Aethina tumida</Emphasis>

Small hive beetle, Aethina tumida Murray, is a parasite of social bee colonies and has become an invasive species, raising concern of the potential threat to native pollinators in its new ranges. Here, we report the defensive behavior strategies used by workers of the Australian stingless bee, Austroplebeia australis Friese, against the small hive beetle. A non-destructive method was used to observe in-hive behavior and interactions between bees and different life stages of small hive beetle (egg, larva, and adult). A number of different individual and group defensive behaviors were recorded. Up to 97% of small hive beetle eggs were destroyed within 90 min of introduction, with a significant increase in temporal rate of destruction between the first and subsequent introductions. A similar result was recorded for 3-day-old small hive beetle larvae, with an increased removal rate from 62.5 to 92.5% between the first and second introductions. Of 32 adult beetles introduced directly into the 4 colonies, 59% were ejected, with the remainder being entombed alive in hives within 6 h. Efficiency of ejection also significantly increased between the first and third introductions. Our observations suggest that A. australis colonies, despite no previous exposure to this exotic parasite, have well developed hive defences that are likely to minimize entry and survival of small hive beetles.     

Behavioral responses of the small hive beetle,Aethina tumida,to odors of three meliponine bee species and honey bees,Apis mellifera scutellata

Recent studies have shown that honey bees, bumble bees, and some meliponine bee species of the genera Trigona, Meliponula, and Dactylurina are hosts of the small hive beetle (SHB) Aethina tumidaMurray (Coleoptera: Nitidulidae), a pest of honey bee colonies in various regions of the world. Olfaction has been implicated in SHB infestations of honey bee and bumble bee colonies. We used olfactometer bioassays to investigate responses of adult male and female SHBs to odors from intact colonies and separate hive components (pot honey, pot pollen, cerumen, and propolis) of three African meliponine bee species, Meliponula ferruginea (Lepeletier) (black morphospecies), M. ferruginea (reddish brown morphospecies), and Meliponula bocandei (Spinola) (Hymenoptera: Apidae). Although both sexes of the beetle strongly preferred intact colony, pot honey, and pot pollen odors, there was no evidence of attraction to propolis and cerumen odors from the three meliponine bee species. Both sexes of SHB also strongly preferred odors from honey bees, Apis mellifera L. (Hymenoptera: Apidae), over odors from the three meliponine bee species. Our results provide substantial evidence of the host potential of African meliponine bees for the SHB, and we discuss this complex association of the SHB with species within the Apidae family.     

An ecological digest of the small hive beetle (Aethina tumida), a symbiont in honey bee colonies (Apis mellifera)

The small hive beetle (Aethina tumida Murray) is an endemic scavenger in colonies of western honey bee subspecies (Apis mellifera L.) inhabiting sub-Saharan Africa where it only occasionally damages host colonies. Such damage is usually restricted to weakened/diseased colonies or is associated with after absconding events (all bees, including the queen, leave the hive) due to behavioral resistance mechanisms of its host. In sharp contrast, the beetle has proven deleterious to honey bee colonies in introduced ranges of the United States and Australia. With this review we synthesize the existing data in a manner that allows us to discuss the beetle’s natural history from an ecological perspective. A thorough exploration of beetle ecology allows us to 1) illuminate the unique symbiotic relationship it and its host share and understand how this relationship is fostered, 2) place this relationship in context with those of other arthropods inhabiting social insect colonies, 3) understand its natural reliance on honey bee colonies, 4) predict its spread outside its native range, and 5) predict its effects on non-African honey bees and non-target species. Here we present an amalgamation of information that will contribute to a more thorough and appropriate understanding of not only small hive beetles as symbionts, but of social insect symbionts in general. Received 4 April 2005; revised 15 October 2005; accepted 18 October 2005.     

Temperature-Dependent Clustering Behavior of Aethina Tumida Murray in Apis Mellifera L. Colonies

Temperate races of honey bees (Apis mellifera) are able to survive cold temperatures by forming thermoregulatory clusters. Small hive beetles (Aethina tumida), which inhabit honey bee colonies in their native range of sub-Saharan Africa and in their introduced ranges of the United States and Australia, are able to endure temperate climates by entering the bee cluster when cold temperatures persist. We conducted an experiment to address the temporal aspects of the cluster-entering behavior of small hive beetles. We did this by exposing beetle-infested observation bee hives to different ambient temperatures and counting the number of beetles remaining in confinement sites on the hive’s periphery at each temperature. The resulting regression analyses suggest that the beetles enter the cluster more rapidly than they exit it, a behavior possibly linked to a colony’s decision to form and dismantle a cluster.     

Small Hive Beetles are Facultative Predators of Adult Honey Bees

Foraging animals can choose to act as predators or not depending on the level of defensiveness of the potential prey. This requires prior evaluation of prey defensiveness, which can be variable, e.g. young insects are usually less able to defend themselves. Here we show that small hive beetles, Aethina tumida, which are scavengers and parasites of honey bee, Apis mellifera, colonies, are facultative predators of young adult host workers. Adult female beetles mounted and attacked young workers more often than their older nestmates, indicating that the beetle is assessing the defensiveness of the host and is adjusting its behaviour accordingly. Since adult female beetles need proteins to activate their ovaries, predation on defenceless young alive host workers offers another rewarding food source, which can obviously not be exploited by beetle larvae. In conclusion, adult small hive beetles seem to be able to assess the trade-off between safety and food reward.     

Olfactory responses of Aethina tumida murray (Coleoptera: Nitidulidae) to some major volatile compounds from hive materials and workers of Apis mellifera

The small hive beetle (Aethina tumida Murray) is an invasive pest affecting honey bee colonies. The beetles are known to be attracted to volatiles from hive products and honey bees like Apis mellifera L. Previously we reported the presence of five major compounds from the volatile extracts of hive materials; ethyl linolenate and ethyl palmitate from pollen dough, oleamide and tetracosane in fermenting honey, and oleamide and 5-methyl-2-phenyl-1H-indole from A. mellifera worker bees. This study tested the attractiveness of the aforementioned five volatile organic compounds to small hive beetles (SHB) by Y-tube olfactometric bioassay. Ethyl linolenate was highly attractive to both male and female adults of SHB. Ethyl palmitate was attractive to SHB only at higher concentration (0.01–01 mg/ml). Interestingly, tetracosane, 5-methyl-2-phenyl-1H-indole and oleamide were repellent for SHB of both sexes, but ethyl linolenate and ethyl palmitate as components of honey bee brood pheromone attracted SHB. The results highlight that SHB differentially utilizes volatile chemicals from hive materials and honey bees as cues to locate honey bee hives.     

Adaptive behaviour of honeybees (Apis mellifera) toward beetle invaders exhibiting various levels of colony integration

Social bees generally host fewer nest invaders than do ants and termites. This is potentially explained by the adaptive defensive strategies of host bees when faced with nest invaders exhibiting various levels of colony integration (based on adaptations to the nest habitat and frequency of nest inhabitation). In the present study, experiments are performed to determine the behaviour at the nest entrance of European honeybee guards Apis mellifera L. (Hymenoptera: Apidae) toward beetle invaders of various levels of behavioural integration into colonies. The species used to test this include Aethina tumida Murray (Coleoptera: Nitidulidae), which is regarded as a highly integrated, unwelcome guest (synechthran) or true guest (symphile); Lobiopa insularis Laporte (Coleoptera: Nitidulidae) and Epuraea luteola Erichson (Coleoptera: Nitidulidae) that are accidentals; and Carpophilus humeralis Fabricius (Coleoptera: Nitidulidae), Carpophilus hemipterus L. (Coleoptera: Nitidulidae) and Tribolium castaneum Herbst (Coleoptera: Tenebrionidae), all of which are species that are not integrated into honeybee colonies. The responses of guard bees to a control bead also are noted. In general, bees ignore T. castaneum and E. luteola to a greater extent than other beetle species. Bees make contact with the black glass bead (a non‐aggressive behaviour) more than they do all beetle species. Bees treat A. tumida more defensively than they treat any other beetle species and the level of bee defensiveness varies by colony. These data suggest an adaptive heightened defensive response by bees toward the most integrated colony intruder but a significantly reduced level of response toward invaders representing all other levels of colony integration.     

Multiple mating by both sexes in an invasive insect species,Aethina tumida (Coleoptera: Nitidulidae)

Multiple mating by both sexes is common among sexually reproducing animals. Small hive beetles (SHB), Aethina tumida, are parasites of bee nests endemic to sub-Saharan Africa and have become a widespread invasive species. Despite the considerable economic damages they can cause, their basic biology remains poorly understood. Here we show that male and female small hive beetles can mate multiple times, suggesting that costs for mating are low in this species. In an invasive A. tumida population in the United States, a combination of laboratory experiments for males and paternity analysis with eight polymorphic DNA microsatellite markers for field-caught females were used to estimate the number of mating by both sexes. The data show that females and males can mate multiple times—females mated with up to eight males, whereas males mated with at least seven females. The results also showed that A. tumida displayed a skewed paternity, although this was not consistent among the tested females. Thus, first or last male advantage seem to be unlikely in A. tumida. Our observations that individuals of both sexes of A. tumida can mate multiple times opens new research avenues for examining drivers of multiple mating and determining the role it may play in promoting biological invasions.     

Infestation of Japanese Native Honey Bees by Tracheal Mite and Virus from Non-native European Honey Bees in Japan

Invasion of alien species has been shown to cause detrimental effects on habitats of native species. Insect pollinators represent such examples; the introduction of commercial bumble bee species for crop pollination has resulted in competition for an ecological niche with native species, genetic disturbance caused by mating with native species, and pathogen spillover to native species. The European honey bee, Apis mellifera, was first introduced into Japan for apiculture in 1877, and queen bees have been imported from several countries for many years. However, its effects on Japanese native honey bee, Apis cerana japonica, have never been addressed. We thus conducted the survey of honey bee viruses and Acarapis mites using both A. mellifera and A. c. japonica colonies to examine their infestation in native and non-native honey bee species in Japan. Honey bee viruses, Deformed wing virus (DWV), Black queen cell virus (BQCV), Israeli acute paralysis virus (IAPV), and Sacbrood virus (SBV), were found in both A. mellifera and A. c. japonica colonies; however, the infection frequency of viruses in A. c. japonica was lower than that in A. mellifera colonies. Based on the phylogenies of DWV, BQCV, and SBV isolates from A. mellifera and A. c. japonica, DWV and BQCV may infect both honey bee species; meanwhile, SBV has a clear species barrier. For the first time in Japan, tracheal mite (Acarapis woodi) was specifically found in the dead honey bees from collapsing A. c. japonica colonies. This paper thus provides further evidence that tracheal-mite-infested honey bee colonies can die during cool winters with no other disease present. These results demonstrate the infestation of native honey bees by parasite and pathogens of non-native honey bees that are traded globally.     

Honey bee colonies from different races show variation in defenses against the varroa mite in a ‘common garden’

Honey bee [Apis mellifera L. (Hymenoptera: Apidae)] genetic diversity may be the key to responding to novel health challenges faced by this important pollinator. In this study, we first compared colonies of four honey bee races, A. m. anatoliaca, A. m. carnica, A. m. caucasica, and A. m. syriaca from Turkey, with respect to honey storage, bee population size, and defenses against varroa. The mite Varroa destructor Anderson & Trueman (Acari: Varroidae) is an important pest of honey bee colonies. There are genetic correlates with two main defenses of bees against this parasite: hygienic behavior, or removing infested brood, and grooming, which involves shaking and swiping off mites and biting them. In the second part of this study, we examined the relationship of these two types of defenses, hygiene and grooming, and their correlation with infestation rates in 32 genetically diverse colonies in a ‘common garden’ apiary. Mite biting was found to be negatively correlated with mite infestation levels.     

Social encapsulation of the small hive beetle (<Emphasis Type="Italic">Aethina tumida</Emphasis> Murray) by European honeybees (<Emphasis Type="Italic">Apis mellifera</Emphasis> L.)

Summary European and African subspecies of honeybees (Apis mellifera L.) utilize social encapsulation to contain the small hive beetle (Aethina tumida Murray), a honeybee colony scavenger. Using social encapsulation, African honeybees successfully limit beetle reproduction that can devastate host colonies. In sharp contrast, European honeybees often fail to contain beetles, possibly because their social encapsulation skills may be less developed than those of African honeybees. In this study, we quantify beetle and European honeybee behaviours associated with social encapsulation, describe colony and time (morning and evening) differences in these behaviours (to identify possible circadian rhythms), and detail intra-colonial, encapsulated beetle distributions. The data help explain the susceptibility of European honeybees to depredation by small hive beetles. There were significant colony differences in a number of social encapsulation behaviours (the number of beetle prisons and beetles per prison, and the proportion of prison guard bees biting at encapsulated beetles) suggesting that successful encapsulation of beetles by European bees varies between colonies. We also found evidence for the existence of circadian rhythms in small hive beetles, as they were more active in the evening rather than morning. In response to increased beetle activity during the evening, there was an increase in the number of prison guard bees during evening. Additionally, the bees successfully kept most (~93%) beetles out of the combs at all times, suggesting that social encapsulation by European honeybees is sufficient to control small populations of beetles (as seen in this study) but may ultimately fail if beetle populations are high.Received 20 January 2003; revised 21 April 2003; accepted 29 April 2003.     

Comparative genomics suggests local adaptations in the invasive small hive beetle

Invasive species are a major driver of ecological and environmental changes that affect human health, food security, and natural biodiversity. The success and impact of biological invasions depend on adaptations to novel abiotic and biotic selective pressures. However, the molecular mechanisms underlying adaptations in invasive parasitic species are inadequately understood. Small hive beetles, Aethina tumida, are parasites of bee nests. Originally endemic to sub‐Saharan Africa, they are now found nearly globally. Here, we investigated the molecular bases of the adaptations to novel environments underlying their invasion routes. Genomes of historic and recent adults A. tumida from both the endemic and introduced ranges were compared. Analysis of gene–environment association identified 3049 candidate loci located in 874 genes. Functional annotation showed a significant bias toward genes linked to growth and reproduction. One of the genes from the apoptosis pathway encodes an “ecdysone‐related protein,” which is a crucial regulator in controlling body size in response to environmental cues for holometabolous insects during cell death and renewal. Genes whose proteins regulate organ size, ovary activation, and oviposition were also detected. Functions of these enriched pathways parallel behavioral differences between introduced and native A. tumida populations, which may reflect patterns of local adaptation. The results considerably improve our understanding of the underlying mechanisms and ecological factors driving adaptations of invasive species. Deep functional investigation of these identified loci will help clarify the mechanisms of local adaptation in A. tumida.     

First Detection of the Larval Chalkbrood Disease Pathogen Ascosphaera apis (Ascomycota: Eurotiomycetes: Ascosphaerales) in Adult Bumble Bees

Fungi in the genus Ascosphaera (Ascomycota: Eurotiomycetes: Ascosphaerales) cause chalkbrood disease in larvae of bees. Here, we report the first-ever detection of the fungus in adult bumble bees that were raised in captivity for studies on colony development. Wild queens of Bombus griseocollis, B. nevadensis and B. vosnesenskii were collected and maintained for establishment of nests. Queens that died during rearing or that did not lay eggs within one month of capture were dissected, and tissues were examined microscopically for the presence of pathogens. Filamentous fungi that were detected were plated on artificial media containing broad spectrum antibiotics for isolation and identification. Based on morphological characters, the fungus was identified as Ascosphaera apis (Maasen ex Claussen) Olive and Spiltoir, a species that has been reported earlier only from larvae of the European honey bee, Apis mellifera, the Asian honey bee, Apis cerana, and the carpenter bee Xylocopa californica arizonensis. The identity of the fungus was confirmed using molecular markers and phylogenetic analysis. Ascosphaera apis was detected in queens of all three bumble bee species examined. Of 150 queens dissected, 12 (8%) contained vegetative and reproductive stages of the fungus. Both fungal stages were also detected in two workers collected from colonies with Ascosphaera-infected B. nevadensis queens. In this study, wild bees could have been infected prior to capture for rearing, or, the A. apis infection could have originated via contaminated European honey bee pollen fed to the bumble bees in captivity. Thus, the discovery of A. apis in adult bumble bees in the current study has important implications for commercial production of bumble bee colonies and highlights potential risks to native bees via pathogen spillover from infected bees and infected pollen.     

Hit‐and‐run trophallaxis of small hive beetles

Some parasites of social insects are able to exploit the exchange of food between nestmates via trophallaxis, because they are chemically disguised as nestmates. However, a few parasites succeed in trophallactic solicitation although they are attacked by workers. The underlying mechanisms are not well understood. The small hive beetle (=SHB), Aethina tumida, is such a parasite of honey bee, Apis mellifera, colonies and is able to induce trophallaxis. Here, we investigate whether SHB trophallactic solicitation is innate and affected by sex and experience. We quantified characteristics of the trophallactic solicitation in SHBs from laboratory‐reared individuals that were either bee‐naïve or had 5 days experience. The data clearly show that SHB trophallactic solicitation is innate and further suggest that it can be influenced by both experience and sex. Inexperienced SHB males begged more often than any of the other groups had longer breaks than their experienced counterparts and a longer soliciting duration than both experienced SHB males and females, suggesting that they start rather slowly and gain more from experience. Successful experienced females and males were not significantly different from each other in relation to successful trophallactic interactions, but had a significantly shorter soliciting duration compared to all other groups, except successful inexperienced females. Trophallactic solicitation success, feeding duration and begging duration were not significantly affected by either SHB sex or experience, supporting the notion that these behaviors are important for survival in host colonies. Overall, success seems to be governed by quality rather than quantity of interactions, thereby probably limiting both SHB energy investment and chance of injury (<1%). Trophallactic solicitation by SHBs is a singular example for an alternative strategy to exploit insect societies without requiring chemical disguise. Hit‐and‐run trophallaxis is an attractive test system to get an insight into trophallaxis in the social insects.     

Identification of the ambrosia fungus of <Emphasis Type="Italic">Xyleborus monographus</Emphasis> and <Emphasis Type="Italic">X. dryographus</Emphasis> (Coleoptera: Curculionidae,Scolytinae)

The scolytid ambrosia beetles Xyleborus monographus and X. dryographus were investigated to identify their nutritional ambrosia fungi. The examination of the oral mycetangia of the beetles, the specialized organs for fungal transport, revealed the dominant occurrence of Raffaelea montetyi, a fungus that was also predominant in the beetle tunnels in the immediate vicinity of the feeding larvae. R. montetyi was previously known only as the ambrosia fungus of the platypodid ambrosia beetle, Platypus cylindrus. These beetle species inhabit the same habitat, mainly trunks of oaks in the Western Palaeartic. The possibility of an exchange of the symbiotic fungus between the ambrosia beetles within their common breeding place is discussed. Consequently, the previous hypothesis of a species-specific association of a single ambrosia fungus with a single beetle species is questioned. A phylogenetic analysis based on DNA sequences classified R. montetyi within the Ophiostomatales of the ascomycetes. The investigation of conidiogenesis of R. montetyi by SEM supported this taxonomic placement and showed the development of the conidia by annellidic percurrent proliferation, identical to the conidiogenesis reported for many anamorph states of the Ophiostomatales.     

Comparative susceptibility and immune responses of Asian and European honey bees to the American foulbrood pathogen,Paenibacillus larvae

American foulbrood (AFB) disease is caused by Paenibacillus larvae. Currently, this pathogen is widespread in the European honey bee— Apis mellifera. However, little is known about infectivity and pathogenicity of P. lan'ae in the Asiatic cavity-nesting honey bees, Apis cerana. Moreover, comparative knowledge of P. larvae infectivity and pathogenicity between both honey bee species is scarce. In this study, we examined susceptibility, larval mortality, survival rate and expression of genes encoding antimicrobial peptides (AMPs) including defensin, apidaecin, abaecin, and hymenoptaecin in A. mellifera and A. cerana when infected with P. larvae. Our results showed similar effects of P. larvae on the survival rate and patterns of AMP gene expression in both honey bee species when bee larvae are infected with spores at the median lethal concentration (LC5 0 ) for A. mellifera. All AMPs of infected bee larvae showed significant upregulation compared with noninfected bee larvae in both honey bee species. However, larvae of A. cerana were more susceptible than A. mellifera when the same larval ages and spore concentration of P. larvae were used. It also appears that A. cerana showed higher levels of AMP expression than A. mellifera. This research provides the first evidence of survival rate, LC50 and immune response profiles of Asian honey bees, A. cerana, when infected by P. larvae in comparison with the European honey bee, A. mellifera.     

Host Specificity in the Honeybee Parasitic Mite,Varroa spp. in Apis mellifera and Apis cerana

The ectoparasitic mite Varroa destructor is a major global threat to the Western honeybee Apis mellifera. This mite was originally a parasite of A. cerana in Asia but managed to spill over into colonies of A. mellifera which had been introduced to this continent for honey production. To date, only two almost clonal types of V. destructor from Korea and Japan have been detected in A. mellifera colonies. However, since both A. mellifera and A. cerana colonies are kept in close proximity throughout Asia, not only new spill overs but also spill backs of highly virulent types may be possible, with unpredictable consequences for both honeybee species. We studied the dispersal and hybridisation potential of Varroa from sympatric colonies of the two hosts in Northern Vietnam and the Philippines using mitochondrial and microsatellite DNA markers. We found a very distinct mtDNA haplotype equally invading both A. mellifera and A. cerana in the Philippines. In contrast, we observed a complete reproductive isolation of various Vietnamese Varroa populations in A. mellifera and A. cerana colonies even if kept in the same apiaries. In light of this variance in host specificity, the adaptation of the mite to its hosts seems to have generated much more genetic diversity than previously recognised and the Varroa species complex may include substantial cryptic speciation.     

Estimating reproductive success of Aethina tumida (Coleoptera: Nitidulidae) in honey bee colonies by trapping emigrating larvae

The small hive beetle (Aethina tumida Murray) is a scavenger and facultative predator in honey bee colonies, where it feeds on pollen, honey, and bee brood. Although a minor problem in its native Africa, it is an invasive pest of honey bees in the United States and Australia. Adult beetles enter bee hives to oviposit and feed. Larval development occurs within the hive, but mature larvae leave the hive to pupate in soil. The numbers leaving, which can be estimated by trapping, measure the reproductive success of adult beetles in the hive over any given period of time. We describe a trap designed to intercept mature larvae as they reach the end of the bottom board on their way to the ground. Trap efficiency was estimated by releasing groups of 100 larvae into empty brood boxes and counting the numbers trapped. Some larvae escaped, but mean efficiency ranged from 87.2 to 94.2%. We envision the trap as a research tool for study of beetle population dynamics, and we used it to track numbers of larvae leaving active hives for pupation in the soil. The traps detected large increases and then decreases in numbers of larvae leaving colonies that weakened and died. They also detected small numbers of larvae leaving strong European and African colonies, even when no larvae were observed in the hives.     

Habitat associations of <Emphasis Type="Italic">Agathidium pulchellum</Emphasis>, an endangered old-growth forest beetle species living on slime moulds

The beetle genus Agathidium is the largest insect group documented that principally feeds on slime moulds. Agathidium pulchellum, one of the rarest Agathidium species in Europe, is listed in the EU’s Habitats Directive. We studied the habitat associations of A. pulchellum in 44 sites located in old-growth and managed forests in eastern Finland. Agathidium pulchellum occurred exclusively on the slime-mould species Trichia decipiens. The host was associated with mid-decayed aspen, spruce and birch logs, and its incidence grew with both increasing log diameter and stand-level log density of spruce and aspen. We also observed that even if its host was present, the beetle was absent from sites with less than 80 aspen and spruce logs per hectare. All sites with A. pulchellum were natural forests of high conservation value. Our results show that it is possible to systematically survey the occurrence of A. pulchellum.