Olfactory response of megachilid bees, Osmia lignaria, Megachile rotundata, and M. pugnata, to individual cues from old nest cavities

The megachilid bees Osmia lignaria Say, Megachile rotundata (F.), and M. pugnata Say were tested for attraction to various components associated with their old nest cavities, or chemical extracts of these components, using a Y-tube olfactory response bioassay. Female bees of these species are known to nest in or near old nest cavities, implying that remnant nest components are important cues for bees looking for nest cavities. Significant results show that female bees were attracted to components that may provide species-specific cues or indicate conspecific nesting activity. Specifically, O. lignaria females showed attraction only to the female cocoon. M. rotundata females were attracted to intact nest cells, the fecal material on the outside of a cocoon, leaf pieces used as nest cell lining, and the extract of leaf pieces. M. pugnata females were attracted to the whole nest cell, the paper straw nesting material with attached cocoon, and feces.     

Experience-dependent plasticity in the mushroom bodies of the solitary bee Osmia lignaria (Megachilidae)

All members of the solitary bee species Osmia lignaria (the orchard bee) forage upon emergence from their natal nest cell. Conversely, in the honey bee, days-to-weeks of socially regulated behavioral development precede the onset of foraging. The social honey bee's behavioral transition to foraging is accompanied by neuroanatomical changes in the mushroom bodies, a region of the insect brain implicated in learning. If these changes were general adaptations to foraging, they should also occur in the solitary orchard bee. Using unbiased stereological methods, we estimated the volume of the major compartments of the mushroom bodies, the neuropil and Kenyon cell body region, in adult orchard bees. We compared the mushroom bodies of recently emerged bees with mature bees that had extensive foraging experience. To separate effects of general maturation from field foraging, some orchard bees were confined to a cage indoors. The mushroom body neuropil of experienced field foragers was significantly greater than that of both recently emerged and mature caged orchard bees, suggesting that, like the honey bee, this increase is driven by outdoor foraging experience. Unlike the honey bee, where increases in the ratio of neuropil to Kenyon cell region occur in the worker after emerging from the hive cell, the orchard bee emerged from the natal nest cell with a ratio that did not change with maturation and was comparable to honey-bee foragers. These results suggest that a common developmental endpoint may be reached via different development paths in social and solitary species of foraging bees.     

Lethal and sublethal effects of imidacloprid on Osmia lignaria and clothianidin on Megachile rotundata (Hymenoptera: Megachilidae)

We examined lethal and sublethal effects of imidacloprid on Osmia lignaria (Cresson) and clothianidin on Megachile rotundata (F.) (Hymenoptera: Megachilidae). We also made progress toward developing reliable methodology for testing pesticides on wild bees for use in pesticide registration by using field and laboratory experiments. Bee larvae were exposed to control, low (3 or 6 ppb), intermediate (30 ppb), or high (300 ppb) doses of either imidacloprid or clothianidin in pollen. Field experiments on both bee species involved injecting the pollen provisions with the corresponding pesticide. Only O. lignaria was used for the laboratory experiments, which entailed both injecting the bee's own pollen provisions and replacing the pollen provision with a preblended pollen mixture containing imidacloprid. Larval development, emergence, weight, and mortality were monitored and analyzed. There were no lethal effects found for either imidacloprid or clothianidin on O. lignaria and M. rotundata. Minor sublethal effects were detected on larval development for O. lignaria, with greater developmental time at the intermediate (30 ppb) and high doses (300 ppb) of imidacloprid. No similar sublethal effects were found with clothianidin on M. rotundata. We were successful in creating methodology for pesticide testing on O. lignaria and M. rotundata; however, these methods can be improved upon to create a more robust test. We also identified several parameters and developmental stages for observing sublethal effects. The detection of sublethal effects demonstrates the importance of testing new pesticides on wild pollinators before registration.     

Olfactory cues and nest recognition in the solitary bee Osmia lignaria

Abstract.  The use of olfactory cues for nest recognition by the solitary bee Osmia lignaria is studied in a greenhouse environment. Glass tubes are provided as nesting cavities to allow the in-nest behaviour of bees to be observed. In addition, each glass tube is cut into three sections for experimental manipulation and for subsequent chemical analysis. Nesting females drag their abdomen along the tube before exiting, spiral inside the tube, and sometimes deposit fluid droplets from the tip of the abdomen. For the manipulation, the outer section, the middle section, or both sections are removed and replaced with similar clean glass tube sections, and the behaviour exhibited by test females is recorded upon arrival in front of the nesting site and inside the nesting tubes. The resulting hesitation behaviour displayed by females after treatments appears to indicate the loss of some olfactory cues used for nest recognition inside the entire nest. Chemical analysis of the depositions inside the nesting tube, as well as analysis of the cuticular lipids of the nesting bees, reveals the presence of free fatty acids, hydrocarbons and wax esters.     

The nectar alkaloid, gelsemine, does not affect offspring performance of a native solitary bee, Osmia lignaria (Megachilidae)

Abstract.  1. The ecology and evolution of foliar-feeding insects are thought to be closely tied to plant secondary compounds. Although secondary compounds are also abundant in floral nectar, their role in mediating pollinator preference and performance remains relatively unexplored.
2. This study tested the effects of an alkaloid, gelsemine, found in the nectar of Carolina jessamine ( Gelsemium sempervirens L., Loganiaceae), on the performance of a native solitary bee ( Osmia lignaria lignaria Say, Megachilidae). Nectar gelsemine reduces visits from pollinators, including O. lignaria lignaria , and gelsemine is toxic to vertebrates and possibly non-native honey bees ( Apis mellifera L., Apidae). To test the hypothesis that the deterrent effects of nectar gelsemine reflect negative consequences for pollinator performance, O. lignaria lignaria offspring provisions were supplemented with nectar containing different gelsemine concentrations. Effects on larval development time, prepupa cocoon mass, adult emergence, and adult mass were measured.
3. Nectar gelsemine had no effect on any measure of offspring performance. Thus, although gelsemine deters foraging by adult bees, this behaviour did not optimize offspring performance under the experimental conditions of this study. In contrast, sugar added to nectar treatments increased offspring mass.
4. While adult pollinators may avoid nectar with secondary compounds, this could hinder offspring performance by reducing sugar in provisions if nectar is limiting in the environment. Preference-performance trade-offs, which are studied extensively with foliar herbivores, have seldom been tested for pollinating plant consumers. Future studies of nectar secondary compounds and insect pollinator preference and performance may help to integrate studies of foliage-consuming insect herbivores with nectar-consuming insect pollinators.     

Different Arginine Transfer Ribonucleic Acid Species Prevalent in Shaken and Unshaken Cultures of Neurospora

When the arginyl-transfer ribonucleic acid (tRNA) species isolated from unshaken and from shaken cultures of Neurospora were compared by co-chromatography, a marked change in the relative abundance of the two main tRNA(arg) species was found. The two arginine tRNA species had different codon responses in ribosome binding assays. The tRNA(arg) eluting first (prevalent in shaken cultures) bound strongly to polyadenylic-guanylic acid [poly(A,G)] and to a lesser extent to polycytidylic-guanylic-adenylic acid [poly(C,G,A)]. The second tRNA(arg) species (prevalent in unshaken cultures) bound to poly(C,G,A) but not to poly(A, G). The possible significance of these observations is briefly discussed. Several modifications that improve the yield of tRNA from Neurospora were introduced in a standard isolation procedure.     

Influence of moisture and aeration on oxygen uptakes in Warburg respiratory experiments with litter and soil

Summary Oxygen uptakes by litter and soil from a beech forest were determined in a Warburg apparatus, using material at field moisture content, at 60 per cent water-holding capacity (WHC), and at saturation (>100 per cent WHC), in unshaken and shaken flasks. In unshaken flasks, oxygen uptakes by materials from some horizons were markedly dependent upon their moisture content. Oxygen uptakes were similar with unshaken materials at 60 per cent WHC and with shaken water-saturated materials; this latter method of determination is preferred for routinely measuring respiratory activity under aerobic conditions. Control of moisture content of the systems appeared necessary to estimate respiratory responses to added solutions of glucose. Significant responses to added glucose were found with materials from all horizons under conditions of adequate moisture and aeration but responses were much less with some unshaken water-saturated materials.     

Evaluation of the shaking technique for the economic management of American foulbrood disease of honey bees (Hymenoptera: Apidae)

Shaking is a nonantibiotic management technique for the bacterial disease American foulbrood (AFB) (Paenibacillus larvae sensu Genersch et al.), in which infected nesting comb is destroyed and the adult honey bees, Apis mellifera L. (Hymenoptera: Apidae), are transferred onto uncontaminated nesting material. We hypothesized that colonies shaken onto frames of uninfected drawn comb would have similar reductions in AFB symptoms and bacterial spore loads than those shaken onto frames of foundation, but they would attain higher levels of production. We observed that colonies shaken onto drawn comb, or a combination of foundation and drawn comb, exhibited light transitory AFB infections, whereas colonies shaken onto frames containing only foundation failed to exhibit clinical symptoms. Furthermore, concentrations of P. larvae spores in honey and adult worker bees sampled from colonies shaken onto all comb and foundation treatments declined over time and were undetectable in adult bee samples 3 mo after shaking. In contrast, colonies that were reestablished on the original infected comb remained heavily infected resulting in consistently high levels of spores, and eventually, their death. In a subsequent experiment, production of colonies shaken onto foundation was compared with that of colonies established from package (bulk) bees or that of overwintered colonies. Economic analysis proved shaking to be 24% more profitable than using package bees. These results suggest that shaking bees onto frames of foundation in the spring is a feasible option for managing AFB in commercial beekeeping operations where antibiotic use is undesirable or prohibited.     

Floating and fidelity in nest visitation byCrawfordapis luctuosa (Hymenoptera: Colletidae)

We studied nest visitation behavior of the ground-nesting beeCrawfordapis luctuosa (Colletidae) at the Monteverde Cloud Forest Preserve, Puntarenas Province, Costa Rica. Some females entered many nests, while others consistently entered only one nest. Multiple nest entry was exhibited by bees we called floaters, and those bees that consistently entered only one nest were provisioning bees. Provisioning bees abandoned their old nests after provisioning periods and became nonprovisioning bees that floated over the nesting area. Floating behavior was a nest-searching behavior typified by oscillating flights just above the ground (<15 cm). Floaters investigated available nests and were not observed to interact aggressively with nest owners. Upon establishing a new nest, floaters became provisioning bees.     

Effect of temperature on Osmia lignaria (Hymenoptera: Megachilidae) prepupa-adult development, survival, and emergence

The solitary bee Osmia lignaria Say continues to be developed as an orchard pollinator in the United States and southern Canada. Female bees are active during the early spring and construct nests consisting of a linear series of unlined cells delimited by mud partitions. Cells are provisioned with a pollen/nectar mass on which an egg is deposited, and nests are sealed with a mud plug. In 1997, we initiated two experiments on the development, mortality, and emergence of O. lignaria at selected laboratory temperature regimes and outdoors. In the first experiment (published previously), we compared temperature treatments for their adequacy in maintaining healthy O. lignaria populations. In a second experiment (reported here), we determined the relationship between rearing temperatures and prepupa-adult development rates as well as emergence time and longevity after wintering and incubation the following spring. We observed important differences in O. lignaria prepupa versus pupa responses to selected temperature treatments. The relationship between temperature and days to pupa was U-shaped, with additional time to transit the prepupa-pupa interval at temperatures above and below 26 degrees C. The negative relationship between temperature and the length of the pupa to adult interval contrasts with the U-shaped thermal response observed for prepupae. Thus, with each increase in thermal heat units over the range of temperature treatments tested, we observed an additional reduction in the pupa-adult interval. Individuals reared at constant 18 degrees C required 2.4 times as many days to transit the pupa-adult interval compared with those at constant 32 degrees C. Our results suggest that there is a need for the development of regionally adapted, latitude-specific breeding populations of this outstanding orchard crop pollinator.     

Induction of mitochondrial migration in the slime mold Physarum polycephalum

Mitochondrial migration in a microplasmodium of Physarum polycephalumwas studied by litgh and electron microscopy. The mitochondriawere dispersed evenly in the microplasmodium of Physarum polycephalumin shaken cultures but when the microplasmodia were left unshakenin a liquid culture for more than 3 hr, the mitochondria migratedtoward the peripheral area and came into contact with an semi-electrontransparent layer beneath the cell membrane. Once the peripherallocalization of mitochondria was established in unshaken culture,subsequent reversal to the shaken cultures induced a reversion.These results suggest that mitochondrial migration is reversiblyindicated by culture condition. (Received June 19, 1978; )     

The importance of odor in nest site selection by a lodger bee, Centris Bicornuta Mocsáry (Hymenoptera: Apidae) in the dry forest of Costa Rica

The more common lodger bee occurring in the dry forest of Costa Rica, Centris bicornuta Muscáry), has been observed nesting in new nest cavities drilled into wooden blocks placed next to cavities used by another female within 2-3 days. In contrast, new nest cavities placed in similar areas with no nesting Centris nearby were not used for weeks. These observations suggest that the presence of nesting bees may play a role in nest site selection. To confirm our observations, new nest cavities were placed in areas with or without nesting. We found nest initiation in newly placed nest cavities only in areas where bees were actively nesting. To examine the possibility that nesting locations are not unique, we placed new nest cavities in new locations either with (a) a number of completed nest cavities or (b) placed alone. Within three days we only found bees nesting in the newly placed nest cavities in situation "a". The results suggested that odor might be involved. We next compared nesting in new cavities placed alone with cavities contaminated with either (a) nest entrance plug material, (b) nest nectar, (c) nest pollen or (d) a combination of pollen and nectar. Nesting was significantly low in cavities placed next to cavities with nest entrance plug material (a), and high in cavities placed next to cavities "b, c, or d". The results suggest that pollen and /or nectar odor play a role in the location of potential nest sites.     

Landmark orientation during the approach to the nest in the stingless beeTrigona (Tetragonisca) angustula (Apidae,Meliponinae)

Summary We displaced a small nest box containing stingless bees (Trigona (Tetragonisca)angustula) over distances of up to 1.6 meters in different directions and counted the numbers of returning foragers to measure the effects of this manipulation on the homing ability of bees. Bees find it hard to locate the nest box when it was displaced more than about 1 m backwards, forwards or sideways relative to the direction into which the nest entrance pointed. They do not find the nest when its height above ground is changed. The bees use landmarks in the vicinity of the nest to locate it: When the nest box is displaced and landmark positions are changed so that their angular position at the new nest site is the same as at the normal nest position their homing ability is less impaired than it is without changes in landmark positions. Our results show that the bees do not use the nest box itself as a landmark until they have approached the nest position to within about 1 meter with the aid of surrounding landmarks.     

Orchard pollination in Capitol Reef National Park,Utah, USA. Honey bees or native bees?

Capitol Reef National Park in central Utah, USA surrounds 22 managed fruit orchards started over a century ago by Mormon pioneers. Honey bees are imported for pollination, although the area in which the Park is embedded has over 700 species of native bees, many of which are potential orchard pollinators. We studied the visitation of native bees to apple, pear, apricot, and sweet cherry over 2 years. Thirty species of bees visited the flowers but, except for pear flowers, most were uncommon compared to honey bees. Evidence that honey bees prevented native bees from foraging on orchard crop flowers was equivocal: generally, honey bee and native bee visitation rates to the flowers were not negatively correlated, nor were native bee visitation rates positively correlated with distance of orchards from honey bee hives. Conversely, competition was tentatively suggested by much larger numbers of honey bees than natives on the flowers of apples, apricots and cherry; and by the large increase of native bees on pears, where honey bee numbers were low. At least one-third of the native bee species visiting the flowers are potential pollinators, including cavity-nesting species such as Osmia lignaria propinqua, currently managed for small orchard pollination in the US, plus several fossorial species, including one rosaceous flower specialist (Andrena milwaukiensis). We suggest that gradual withdrawal of honey bees from the Park would help conserve native bee populations without decreasing orchard crop productivity, and would serve as a demonstration of the commercial value of native pollinators.     

Old comb for nesting site recognition by <Emphasis Type="Italic">Apis dorsata?</Emphasis> Field experiments in China

The Asian giant honey bee, Apis dorsata, often conducts seasonal, long-distance migrations in southern China, between a preferred tree (having more than one nest) and alternate sites. Although worker bees cannot make a round-trip journey, colonies re-utilize preferred trees after an absence of several months. We performed comb experiments in which bases and all abandoned combs were entirely scraped off trees and their sites covered with plastic, or comb was moved to trees of the same species. Swarms of giant honey bees investigated trees where combs were removed and continued to nest on the same trees. In contrast, placing combs in nets on previously used trees, or on nearby trees of the same species, did not attract more swarms. The same number of colonies that left them returned to previously occupied trees. Our findings suggest that direct olfactory or sensory contact with old comb bases might regulate nest establishment, but individual trees, lacking normal visual or chemical cues of old nests, are relocated using behavioral devices that remain to be elucidated. Received 12 February 2007; revised 5 June 2007; accepted 13 September 2007.     

Individual bumblebees vary in response to disturbance: a test of the defensive reserve hypothesis

Bees may leave their nest in the event of an attack, but this is not their only response. Here, we examine the behavior of those individuals that remain inside the nest during a disturbance. Specifically, we test the hypothesis that bee workers usually exhibiting high levels of inactivity (i.e., ‘lazy’ bees) may function as defensive reserves that are more likely to respond when the colony is disturbed. We explore this hypothesis by simulating vertebrate attacks by vibrating or blowing carbon dioxide into two colonies on alternating days and measuring the movements and tasks performed by bees inside the nest. Our results show that regardless of the disturbance type, workers increase guarding behavior after a disturbance stops. Although previously inactive bees increased their movement speed inside the nest when the disturbance was vibration, they were not more likely to leave the nest (presumably to attack the simulated attacker) or switch to guarding behavior for any disturbance type. We therefore reject the hypothesis that inactive Bombus impatiens bumblebees act as defensive reserves, and propose alternative hypotheses regarding why many workers remain inactive inside the nest.     

PVC nest boxes are less at risk of occupancy by feral honey bees than timber nest boxes and natural hollows

Feral European Honey Bee (Apis mellifera) has been identified as a potential nest competitor for Australian hollow nesting species, but few studies have investigated the impact of feral honey bee competition on Threatened species. Our study used data from Glossy Black‐cockatoo (Calyptorhynchus lathami halmaturinus) nests on Kangaroo Island, monitored and managed over an 11‐year period, and found 12% of nests became occupied by feral honey bees during that period. Our results indicate that feral honey bees were less likely to occupy nest boxes made of PVC (5%) compared with wooden nest boxes (24%) or natural hollows in Eucalyptus trees (14%). The removal of feral honey bee hives from nests is a priority for long‐term conservation of glossy black‐cockatoos on Kangaroo Island. We recommend that PVC nest boxes are chosen for future nesting habitat restoration, due to the more frequent use of wooden nest boxes by feral honey bees.     

Nesting aggregation as a predictor of brood parasitism in mason bees (Osmia spp.)

1. Parasitism can be an important source of mortality for insect populations; however, we know little about the factors influencing vulnerability of wild bees to parasites. Mason bees (genus Osmia; Hymenoptera: Megachilidae) are important pollinators of crops and wild plants and are vulnerable to attack by brood parasites. High nest densities may increase rates of brood parasitism by attracting disproportionate numbers of parasites. 2. Three years of field observations from multiple sites were analysed to assess whether mason bee brood parasitism increased with host density. Mason bees were allowed to nest in artificial nesting blocks and establish natural variation in nesting density. Nest cells constructed by bees were checked for the presence of parasite eggs. 3. Parasitism of nest cells strongly increased with the number of actively nesting bees at a nesting block. Mason bees showed no preference for nesting in blocks that were occupied or unoccupied by other mason bees. Parasitism also increased with the number of days a nest was provisioned and decreased over the course of the season. Nest cells constructed last in a nest were significantly more parasitised than inner cells, despite being sealed against invasions. 4. These findings show positively density‐dependent parasitism in mason bees. They also suggest that bees terminate parasitised nests, causing parasitised cells to become outermost nest cells – a behaviour that may represent a defence against parasites. Our results have implications for the management of mason bees as agricultural pollinators, as cultivating them at high densities could reduce offspring survival.     

Ground-nesting bees determine the location of their nest relative to a landmark by other than angular size cues

Bees and wasps acquire a visual representation of their nest's environment and use it to locate their nest when they return from foraging trips. This representation contains among other features cues to the distance of near-by landmarks. We worked with two species of ground-nesting bees, Lasioglossum malachurum (Hymenoptera: Halictidae), Dasypoda hirtipes (Hymenoptera: Melittidae) and asked which cues to landmark distance they use during homing. Bees learned to associate a single cylindrical landmark with their nest's location. We subsequently tested returning bees with landmarks of different sizes and thus introduced large discrepancies between the angular size of the landmark as seen from the nest during training and its distance from the nest. The bees' search behaviour and their choice of dummy nest entrances show that both species of ground-nesting bees consistently search for their nest at the learned distance from landmarks. The influence of the apparent size of landmarks on the bees' search and choice behaviour is comparatively weak. We suggest that the bees exploit cues derived from the apparent speed of the landmark's image at their retina for distance evaluation.     

Visual marks learned by the solitary bee Megachile rotundata for localizing its nest

We studied homing behaviour of leaf-cutter bees, Megachile rotundata, by using artificial landmarks. We evaluated their nest-searching behaviour in different test situations to elucidate the nature of the visual marks they used in this task. When we modified or removed geometrical figures surrounding the nest, the bees searched for longer, showing that they noticed the introduced changes. However, these manipulations never prevented bees from finding their nest, suggesting that other visual cues were crucial in the task. Manipulations of the edges provided by the boundaries of the device (nest block, metal sheet on which the block was mounted) strongly impaired the homing performance. The further away the edges that were left intact, the stronger was the impairment of the homing behaviour. These results suggest that bees learn the distances of the various edges from the goal and that edges have a hierarchical significance according to their distance from the nest. The most distant edges provide vague information, which suffices to guide the insect towards the next edge in the sequence, until it recognizes the final, precise location of the nest. The results support the conclusion that information on distances is acquired using cues derived from motion parallax generated by the insect's self-motion. Recognition of edge parameters such as position and orientation might be achieved by an image-matching mechanism based on dynamic processes. Thus, in the homing task, there is no clear discrepancy between the eidetic and the parametric hypotheses of spatial representation.