Culturing chelifers (Pseudoscorpions) that consume Varroa mites

Chelifers (Pseudoscorpions) are generalist predators of small prey such as mites. Their occasional presence in honeybee hives suggests potential to exploit them as part of a management programme against Varroa mites (Varroa destructor), a significant pest of honeybees. Two species of native New Zealand chelifers Nesochernes gracilis and Heterochernes novaezealandiae, shown to consume Varroa mites, were collected from commercial nucleus hives or in litter surrounding the hives. Methods for mass‐rearing the chelifers were developed to provide specimens for research and introduction into beehives for biological control of Varroa. Cultures were fed aphids and fruit fly larvae in vented containers containing sand and bark. N. gracilis was maintained at 14°C, 18°C, and 22°C. At 18°C, 1423 nymphs were reared from 140 N. gracilis adults, with 84.8% of all nymphs produced at this temperature. H. novaezealandiae was maintained at 18°C and 22°C, with 5 nymphs raised from 12 adults at 18°C and none at the higher temperature.     

Heavy Metal (Hg,Cr, Cd,and Pb) Contamination in Urban Areas and Wildlife Reserves: Honeybees as Bioindicators

The degree of heavy metal (Hg, Cr, Cd, and Pb) pollution in honeybees (Apis mellifera) was investigated in several sampling sites around central Italy including both polluted and wildlife areas. The honeybee readily inhabits all environmental compartments, such as soil, vegetation, air, and water, and actively forages the area around the hive. Therefore, if it functions in a polluted environment, plant products used by bees may also be contaminated, and as a result, also a part of these pollutants will accumulate in the organism. The bees, foragers in particular, are good biological indicators that quickly detect the chemical impairment of the environment by the high mortality and the presence of pollutants in their body or in beehive products. The experiment was carried out using 24 colonies of honeybees bred in hives dislocated whether within urban areas or in wide countryside areas. Metals were analyzed on the foragers during all spring and summer seasons, when the bees were active. Results showed no presence of mercury in all samples analyzed, but honeybees accumulated several amounts of lead, chromium, and cadmium. Pb reported a statistically significant difference among the stations located in urban areas and those in the natural reserves, showing the highest values in honeybees collected from hives located in Ciampino area (Rome), next to the airport. The mean value for this sampling station was 0.52 mg kg⁻¹, and July and September were characterized by the highest concentrations of Pb. Cd also showed statistically significant differences among areas, while for Cr no statistically significant differences were found.     

Effects of Hierarchical Roost Removal on Northern Long-Eared Bat (Myotis septentrionalis) Maternity Colonies

Forest roosting bats use a variety of ephemeral roosts such as snags and declining live trees. Although conservation of summer maternity habitat is considered critical for forest-roosting bats, bat response to roost loss still is poorly understood. To address this, we monitored 3 northern long-eared bat (Myotis septentrionalis) maternity colonies on Fort Knox Military Reservation, Kentucky, USA, before and after targeted roost removal during the dormant season when bats were hibernating in caves. We used 2 treatments: removal of a single highly used (primary) roost and removal of 24% of less used (secondary) roosts, and an un-manipulated control. Neither treatment altered the number of roosts used by individual bats, but secondary roost removal doubled the distances moved between sequentially used roosts. However, overall space use by and location of colonies was similar pre- and post-treatment. Patterns of roost use before and after removal treatments also were similar but bats maintained closer social connections after our treatments. Roost height, diameter at breast height, percent canopy openness, and roost species composition were similar pre- and post-treatment. We detected differences in the distribution of roosts among decay stages and crown classes pre- and post-roost removal, but this may have been a result of temperature differences between treatment years. Our results suggest that loss of a primary roost or ≤ 20% of secondary roosts in the dormant season may not cause northern long-eared bats to abandon roosting areas or substantially alter some roosting behaviors in the following active season when tree-roosts are used. Critically, tolerance limits to roost loss may be dependent upon local forest conditions, and continued research on this topic will be necessary for conservation of the northern long-eared bat across its range.     

Biomonitoring with Honeybees of Heavy Metals and Pesticides in Nature Reserves of the Marche Region (Italy)

The aim of this study was to carry out biomonitoring with honeybees (Apis mellifera L.) to assess the presence of pesticides and heavy metals (cadmium, chromium, nickel, lead) in all of the ten nature reserves of the Marche Region (central–eastern Italy). The study was carried out during the spring and summer seasons when the honeybees were active, over 3 years (2008–2010). Twenty-two colonies of honeybees bred in hives were used. Samples of live and dead honeybees and of honey were collected from 11 sampling stations from May to October in each year. No pesticide pollution was found. Significant differences in heavy metal concentrations were found among years, months and sites, and in particular situations. The analysis reveals that high heavy-metal concentrations occurred exclusively in live honeybees. For the seasonal averages, the most detected heavy metal was chromium, which exceeded the threshold more often than for the other elements, followed by cadmium and lead; nickel never exceeded the threshold. The data are discussed with an evaluation of the natural and anthropic sources taken from the literature and from local situations that were likely to involve heavy metal pollution.     

A DNA barcoding method for identifying and quantifying the composition of pollen species collected by European honeybees, <Emphasis Type="Italic">Apis mellifera</Emphasis> (Hymenoptera: Apidae)

The European honeybee, Apis mellifera L. (Hymenoptera: Apidae), is the most important crop pollinator, and there is an urgent need for a sustained supply of honeybee colonies. Understanding the availability of pollen resources around apiaries throughout the brood-rearing season is crucial to increasing the number of colonies. However, detailed information on the floral resources used by honeybees is limited due to a scarcity of efficient methods for identifying pollen species composition. Therefore, we developed a DNA barcoding method for identifying the species of each pollen pellet and for quantifying the species composition by summing the weights of the pellets for each species. To establish the molecular biological protocol, we analyzed 1008 pellets collected between late July and early September 2016 from five hives placed in a forest/agricultural landscape of Hokkaido, northern Japan. Pollen was classified into 31 plant taxa, of which 29 were identified with satisfactory discrimination (25 species and 4 genera) using trnL-trnF and ITS2 as DNA barcoding regions together with available floral and phenological information. The remaining two taxa were classified to the species level using other DNA barcoding regions. Of the 1008 pollen pellets tested, 1005 (99.7%) were successfully identified. As an example of the use of this method, we demonstrated the change in species composition of pollen pellets collected each week for 9 weeks from the same hive.     

Pollination disruption by European honeybees in the Australian bird-pollinated shrubGrevillea barklyana (Proteaceae)

European honeybees (Apis mellifera) were less efficient pollinators ofGrevillea barklyana than nectar-feeding birds. Nectar-collecting honeybees did not contact reproductive parts of flowers. Pollen-collecting honeybees preferentially visited malestage flowers but rarely visited female-stage flowers. Fruit set on caged inflorescences that allowed access to honeybees but excluded birds was reduced by more than 50% compared to inflorescences that were visited by both types of visitors. Further, fruit set on caged inflorescences was less than on bagged inflorescences that excluded both birds and honeybees, indicating that pollen removal by bees decreased opportunities for delayed autonomous selfing in the absence of birds. Although fruit set was not pollen-limited at the study site, pollen removal by honeybees would decrease fruit set in small populations where birds are scarce. In addition, pollen removal by honeybees would reduce opportunities for outcrossing and reproductive success through male function. Although honeybees have been in Australia for insufficient time to have exerted selection on floral traits, evolutionary shifts in response to these animals are likely to occur in the future.     

Integrated crop pollination to buffer spatial and temporal variability in pollinator activity

Insect pollination improves the yield and quality of many crops, yet there is increasing evidence of insufficient insect pollinators limiting crop production. Effective Integrated Crop Pollination (ICP) involves adaptable, targeted and cost-effective management of crop pollination and encourages the use of both wild and managed pollinators where appropriate. In this study we investigate how the addition of honeybee hives affects the community of insects visiting oilseed rape, and if hive number and location affect pollinator foraging and oilseed rape pollination in order to provide evidence for effective ICP. We found that introducing hives increased overall flower visitor numbers and altered the pollinator community, which became dominated by honeybees. Furthermore a greater number of hives did not increase bee numbers significantly but did result in honeybees foraging further into fields. The timing of surveys and proximity to the field edge influenced different pollinators in different ways and represents an example of spatial and temporal complementarity. For example dipteran flower visitor numbers declined away from the field edge whereas honeybees peaked at intermediate distances into the field. Furthermore, no significant effects of survey round on wild bees overall was observed but honeybee numbers were relatively lower during peak flowering and dipteran abundance was greater in later survey rounds. Thus combining diverse wild pollinators and managed species for crop pollination buffers spatial and temporal variation in flower visitation. However we found no effect of insect pollination on seed set or yield of oilseed rape in our trial, highlighting the critical need to understand crop demand for insect pollination before investments are made in managing pollination services.     

Duration and spread of an entomopathogenic fungus, Beauveria bassiana (Deuteromycota: Hyphomycetes), used to treat varroa mites (Acari: Varroidae) in honey bee (Hymenoptera: Apidae) hives

A strain of the fungus Beauveria bassiana (Balsamo) Vuillemin (Deuteromycota: Hyphomycetes) isolated from varroa mites, Varroa destructor Anderson & Trueman (Acari: Varroidae), was used to treat honey bees, Apis mellifera L. (Hymenoptera: Apidae), against varroa mites in southern France. Fungal treatment caused a significant increase in the percentage of infected varroa mites compared with control treatments in two field experiments. In the first experiment, hives were treated with a formulation containing 0.37 g of B. bassiana conidia per hive and in the second experiment with a dose of 1.0 g of conidia per hive. The percentage of infected varroa mites also increased in the nontreated (control) hives, suggesting a movement of conidia, probably via bee drift, among the hives. Mite fall was significantly higher among treated hives compared with control hives on the sixth and eighth days after treatment in the first experiment. These days correspond to previously published data on the median survivorship of mites exposed to that fungal solate. The interaction of treatment and date was significant in the second experiment with respect to mite fall. Increases in colony-forming unit (cfu) density per bee were observed in all treatments but were significantly higher among bees from treated hives than control hives for at least a week after treatment. The relationship between cfu density per bee and proportion infected was modeled using a sigmoid curve. High levels of infection (>80%) were observed for cfu density per bee as low as 5 x 102 per bee, but the cfu density in hives treated with 0.37 g generally dropped below this level less than a week after treatment.     

Range and Frequency of Africanized Honey Bees in California (USA)

Africanized honey bees entered California in 1994 but few accounts of their northward expansion or their frequency relative to European honey bees have been published. We used mitochondrial markers and morphometric analyses to determine the prevalence of Africanized honeybees in San Diego County and their current northward progress in California west of the Sierra Nevada crest. The northernmost African mitotypes detected were approximately 40 km south of Sacramento in California’s central valley. In San Diego County, 65% of foraging honey bee workers carry African mitochondria and the estimated percentage of Africanized workers using morphological measurements is similar (61%). There was no correlation between mitotype and morphology in San Diego County suggesting Africanized bees result from bidirectional hybridization. Seventy percent of feral hives, but only 13% of managed hives, sampled in San Diego County carried the African mitotype indicating that a large fraction of foraging workers in both urban and rural San Diego County are feral. We also found a single nucleotide polymorphism at the DNA barcode locus COI that distinguishes European and African mitotypes. The utility of this marker was confirmed using 401 georeferenced honey bee sequences from the worldwide Barcode of Life Database. Future censuses can determine whether the current range of the Africanized form is stable, patterns of introgression at nuclear loci, and the environmental factors that may limit the northern range of the Africanized honey bee.     

Honeybees have Hydrophobic Wings that Enable Them to Fly through Fog and Dew

Honeybees have received public attention for their remarkable performance in low-altitude flying and their outstanding airborne hovering capability.However,minimal attention has been given to their capability to fly through the harshest climatic conditions.In this study,we used a high-speed camera and recorded an interesting phenomenon in which honeybees (Apis mellifera ligustica) flew effortlessly through mists or drizzling rain.To identify the mechanism behind honeybees flying through mists,the microstructure of their wings was examined via atomic force microscopy and scanning electron microscopy.Experimental results showed that the surface of a honeybee wing was rough,with bristles distributed on both the dorsal and ventral sides.The measurement results of the contact angle proved that the surface of honeybee wings was hydrophobic.Furthermore,hydrophobic proteins,which contained at least one hydrophobic tetra-peptide (i.e.,AAPA/V),were obtained.The rugged surface and hydrophobic proteins caused the hydrophobicity of honeybee wings.These results identify the hydrophobic mechanism of honeybee wings,which will be useful in designing hydrophobic structures.     

Effects of landscape metrics on Apis and non-Apis pollinators and seed set in common buckwheat

Crop pollination is an essential ecosystem service, which, when provided by wild pollinator species, can be negatively affected by human activities at a landscape scale. Various pollinator species can be affected by landscape factors and/or act at different spatial scales. We investigated whether different landscape factors at different spatial scales are correlated with pollinator abundances and pollination success. We examined common buckwheat, which is a self-incompatible crop species, pollinated by managed honeybees, native honeybees, and wild non-honeybee insects. To determine the effects of landscape factors at different spatial scales, we sampled flower-visiting insects and recorded seed sets. Results from the sampled flower visitors indicated that the abundances of managed honeybees, native honeybees, and wild non-honeybee insects were significantly related to the distance of managed hives, the area of forest cover within a 3-km radius, and the area of forest and grassland cover within a 100-m radius, respectively. Additionally, the latter two influential factors of the landscape, but not the distance of managed hives, showed significant positive correlations with the seed set. These results suggest that considering landscape factors at different spatial scales associated with the characteristics of managed and wild pollinators would help to ensure crop yields.     

How do honeybees attract nestmates using waggle dances in dark and noisy hives?

It is well known that honeybees share information related to food sources with nestmates using a dance language that is representative of symbolic communication among non-primates. Some honeybee species engage in visually apparent behavior, walking in a figure-eight pattern inside their dark hives. It has been suggested that sounds play an important role in this dance language, even though a variety of wing vibration sounds are produced by honeybee behaviors in hives. It has been shown that dances emit sounds primarily at about 250-300 Hz, which is in the same frequency range as honeybees' flight sounds. Thus the exact mechanism whereby honeybees attract nestmates using waggle dances in such a dark and noisy hive is as yet unclear. In this study, we used a flight simulator in which honeybees were attached to a torque meter in order to analyze the component of bees' orienting response caused only by sounds, and not by odor or by vibrations sensed by their legs. We showed using single sound localization that honeybees preferred sounds around 265 Hz. Furthermore, according to sound discrimination tests using sounds of the same frequency, honeybees preferred rhythmic sounds. Our results demonstrate that frequency and rhythmic components play a complementary role in localizing dance sounds. Dance sounds were presumably developed to share information in a dark and noisy environment.     

Plant essential oils as potential control agents of varroatosis

Beekeeping has always been vulnerable to various sanitary drawbacks. The mite Varroa destructor (Mesostigmata: Varroidae), an obligated ectoparasite of honeybees, has been in recent times one of the major problems leading not only to economical losses but also to ecological problems related to the role of honeybees as the most important pollinators on Earth. Varroatosis has been treated by methods that include special practices of beekeeping, physical removal, and the use of synthetic acaricides. None of these methods have, however, rendered complete protection, and there is clear evidence for the evolution of resistance to conventional acaricides. Consequently, the need for alternative control means has prompted the onset of several prospecting programs on botanicals with anti-varroa potential. Among these, essential oils appear as good candidates for safer control. In this review, we provide a concise compilation of the information generated in the last years on essential oils with anti-varroa activity. Future developments in this area demand standardization of bioassay protocols (either in the laboratory for primary screening or in hives), a detailed chemical characterization of the essential oils tested, and comprehensive studies on application methods.     

青阳参花部特征及其传粉适应性

对青阳参花（Cynanchum otophyllum）部综合特征、访花昆虫种类、访花行为及传粉过程进行了研究,结果表明,青阳参花结构复杂,两个子房基部离生、花柱联合与雄蕊形成合蕊柱,柱头表面被邻近花药的侧翼紧密包围形成5个柱头腔。青阳参的花粉形成独特的花粉块,一次传粉过程可以转运大量的花粉。东方蜜蜂（Apis cerana）是青阳参的主要传粉昆虫,其传粉包括两个过程：（1）当蜜蜂的口器或足插入着粉腺的槽口后借助蜜蜂的力量将花粉块从花上拔起;（2）当蜜蜂再次访花时将携带的花粉块插入其中一个柱头腔。花粉块里面的花粉粒住柱头腔中萌发出花粉管,然后沿着花柱道向下生长最后进入子房。在整个花期仡粉保持有相对较高的生活力,而其柱头可授性则在7天后逐渐降低。     

Thermoregulation during flight: body temperature and sensible heat transfer in free-ranging Brazilian free-tailed bats (Tadarida brasiliensis)

Bat wings are important for thermoregulation, but their role in heat balance during flight is largely unknown. More than 80% of the energy consumed during flight generates heat as a by-product, and thus it is expected that bat wings should dissipate large amounts of heat to prevent hyperthermia. We measured rectal (T(r)) and surface (T(s)) temperatures of Brazilian free-tailed bats (Tadarida brasiliensis) as they emerged from and returned to their daytime roosts and calculated sensible heat transfer for different body regions (head, body, wings, and tail membrane). Bats' T(r) decreased from 36.8°C during emergence flights to 34.4°C during returns, and T(s) scaled positively with ambient temperature (T(a)). Total radiative heat loss from bats was significantly greater for a radiative sink to the night sky than for a sink with temperature equal to T(a). We found that free-ranging Brazilian free-tailed bats, on average, do not dissipate heat from their wings by convection but instead dissipate radiative heat (L) to the cloudless night sky during flight ([Formula: see text] W). However, within the range of T(a) measured in this study, T. brasiliensis experienced net heat loss between evening emergence and return flights. Regional hypothermia reduces heat loss from wings that are exposed to potentially high convective fluxes. Additional research is needed to establish the role of wings in evaporative cooling during flight in bats.     

Roost selection by Formosan leaf-nosed bats (Hipposideros armiger terasensis)

Patterns of roost use by Formosan leaf-nosed bats (Hipposideros armiger terasensis) were studied from November 1998 to April 2000. Structural characteristics, microclimates, and disturbance levels of 17 roosts used by H. a. terasensis and 15 roosts either used by other bat species (2) or not occupied by any bat species were compared. Roosts used by these bats were significantly larger in size and had greater areas covered by water compared to unused roosts. Entrances of active roosts were more likely to be east-west oriented. Hibernacula had lower entrances and ceilings than did roosts used only in summer. Higher temperatures were recorded in non-breeding roosts than in breeding roosts, but temperature gradients in these two types of roosts did not differ. In winter, hibernacula were warmer, and the temperature fluctuated less than in non-hibernacula. The relative humidities in summer roosts and hibernacula were nearly 100%. Disturbance levels were significantly higher in non-breeding roosts than in breeding roosts, and in non-hibernacula than in hibernacula. These results suggest that the Formosan leaf-nosed bats are selective of their roosts, but the pattern of their roost selection differs from those reported for bats of temperate regions. The reasons for such differences may be related to differences in body size, behavior, and reproductive strategy of the Formosan leaf-nosed bats living in a subtropical climate in Taiwan.     

An Amino Acid Substitution (L925V) Associated with Resistance to Pyrethroids in Varroa destructor

The Varroa mite, Varroa destructor, is an important pest of honeybees and has played a prominent role in the decline in bee colony numbers over recent years. Although pyrethroids such as tau-fluvalinate and flumethrin can be highly effective in removing the mites from hives, their intensive use has led to many reports of resistance. To investigate the mechanism of resistance in UK Varroa samples, the transmembrane domain regions of the V. destructor voltage-gated sodium channel (the main target site for pyrethroids) were PCR amplified and sequenced from pyrethroid treated/untreated mites collected at several locations in Central/Southern England. A novel amino acid substitution, L925V, was identified that maps to a known hot spot for resistance within the domain IIS5 helix of the channel protein; a region that has also been proposed to form part of the pyrethroid binding site. Using a high throughput diagnostic assay capable of detecting the mutation in individual mites, the L925V substitution was found to correlate well with resistance, being present in all mites that had survived tau-fluvalinate treatment but in only 8 % of control, untreated samples. The potential for using this assay to detect and manage resistance in Varroa-infected hives is discussed.     

Evidence for competition between honeybees and bumblebees; effects on bumblebee worker size

Numerous studies suggest that honeybees may compete with native pollinators where introduced as non-native insects. Here we examine evidence for competition between honeybees and four bumblebee species in Scotland, a region that may be within the natural range of honeybees, but where domestication greatly increases the honeybee population. We examined mean thorax widths (a reliable measure of body size) of workers of Bombus pascuorum, B. lucorum, B. lapidarius and B. terrestris at sites with and without honeybees. Workers of all four species were significantly smaller in areas with honeybees. We suggest that reduced worker size is likely to have implications for bumblebee colony success. These results imply that, for conservation purposes, some restrictions should be considered with regard to placing honeybee hives in or near areas where populations of rare bumblebee species persist.     

Further investigations on the value of electric heating of beehives

A set of twenty-four colonies of honeybees was divided into three groups, two of which received different intensities of artificial heat, applied by means of specially designed heating frames placed in the middle of the brood chambers, from 2 Jan. to 16 Mar. 1942. Heating the colonies reduced the area of brood present at the beginning of May without noticeably affecting the consumption of carbohydrate stores and thus had an adverse effect on the colonies. This conclusion is in agreement with that reached as a result of earlier work (Butler & Cockbill, 1942.)
Colonies in hives fitted with top entrances were no stronger in the spring than colonies in hives with bottom entrances, nor did they show any significant difference in store consumption.
No significant brood chamber temperature differences were found in hives fitted with top entrances as compared with control hives which had normal bottom entrances.
During the period when heat was applied to the colonies by means of the heating frames the minimum brood chamber temperature was significantly higher in the heated hives than in the control, unheated, hives, this being especially marked in the case of the hives fitted with 6 W. heaters.
Throughout the experiment the minimum brood chamber temperature of all the hives showed a close dependence on screen temperature. During the preliminary period before the heat was turned on a change of 1 C. screen temperature was associated with change of 0.92 C. in the minimum brood chamber temperature in hives with top entrances, and a change of 0.86 C. in hives with bottom entrances. After the heat was turned on the corresponding figures were 0.84 C. for top entrance hives and 0.67 C. for the bottom entrance hives.     

Efficiency of essential oil (Satureja montana) in controlling the ascospherosis in the honey bee (Apis mellifera) under field conditions.

With the aim of testing its effect in the control of ascosphaerosis in bees, the essential oil of ajedra was incorporated into three types of feed in five different concentrations. Syrup (water and honey) with 0.1% were the best tolerated by the bees, with no colonial changes after a single feed. Later, the ascosphaerosis was introduced in a controlled manner into eight hives. For this, a 10 e6/ml spore suspension was applied by nedulisor three times a week for four weeks to ensure its abundant presence inside the hives. This was continued for the next four weeks period, and at the same time portions of brood-comb in a known state (24 h before operculation) were removed from the hives and heat shocked for 24 h (22 -/+ 2 degrees C), then replaced in the hives for their opercultion, finally removing them and maintaining them at 35 degrees C and 70% relative humidity until the appearance of typical disease symptoms (mummification of the larvae).The hives were randomly divided into two groups of four. One group received 500 ml of syrup with 0.1% oil of ajedra, twice a week for four weeks. The other was kept as the control, receiving syrup without medication. 27.6% of the selected larvae in the treated hives showed mummification, compared with 79.1% in the control hives. The treatment was perfectly tolerated by the bee colonies, showing no changes in their subsequent development.