Influence of rough handling on Osmia lignaria (Hymenoptera: Megachilidae) nest establishment in commercial orchards

Osmia lignaria Say (Hymenoptera: Megachilidae) can be used to pollinate fruit trees. Populations are sometimes difficult to sustain because some female bees fail to establish at provided nesting sites. We address the hypothesis that rough handling of overwintered O. lignaria results in decreased establishment. We tested this by shaking (200 rpm for 2 min) overwintering bees as a proxy for rough handling. Bees were then released in an orchard, and nest establishment of shaken and unshaken bees was recorded. There was no significant difference in the proportion of shaken and unshaken females that nested, indicating that rough handling of overwintering bees does not discourage nest establishment.     

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 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.     

Foraging and nesting behavior of Osmia lignaria (Hymenoptera: Megachilidae) in the presence of fungicides: cage studies

During orchard pollination studies in California, we observed dramatic changes in nesting and foraging behavior of Osmia lignaria Say (Hymenoptera: Megachilidae) after sprays with tank mixtures containing fungicides. A characteristic pattern of postspray events observed includes erratic behavior and interrupted foraging and nesting activity for several days. In an effort to determine whether fungicidal sprays were disruptive to bee foraging and thus to pollination, we exposed O. lignaria females nesting in field cages planted with lacy scorpionweed, Phacelia tanacetifolia Benth (Hydrophyllaceae), to selected spray mixtures normally encountered in California orchard production systems: iprodione (Rovral), propiconazole (Orbit), benomyl (Benlate), and captan (Captan 50 WP); the surfactant Dyne-Amic, alone and mixed with Rovral; and the tank mixture IDB (Rovral + Dyne-Amic + the foliar fertilizer Bayfolan Plus). An additional cage sprayed with an equal volume of water acted as control, and a cage sprayed with the insecticide dimethoate as a toxic standard. For each female O. lignaria, we recorded time spent inside the nest depositing pollen-nectar loads, foraging time, cell production rate, and survival. All females in the dimethoate treatment died postspray + 1 d. Before death, some of these females behaved similarly to our previous orchard observations. A high proportion of females in the IDB cage were inactive for a few hours before resuming normal foraging and nesting activity. No lethal or behavioral effects were found for any of the other compounds or mixtures tested. Our results indicate that the fungicide applications that we tested are compatible with the use of O. lignaria as an orchard pollinator.     

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.     

Spatial Nest‐Settlement Decisions in Digger Wasps: Conspecifics Matter more than Heterospecifics and Previous Experience

Research into the driving forces behind spatial arrangement of wasp nests has considered abiotic environmental factors, but seldom investigated attraction or repulsion towards conspecifics or heterospecifics. Solitary female digger wasps (Hymenoptera) often nest in dense aggregations, making these insects good models to study this topic. Here, we analysed the nesting patterns in an area shared by three species of the genus Bembix, in a novel study to discover whether female wasps are attracted to or repulsed by conspecific nests, heterospecific nests or their own previously established nests when choosing nest‐digging locations. Early in the season, each species showed a clumping pattern of nests, but later in the season, a random distribution of nests was more common, suggesting an early conspecific attraction. Such behaviour was confirmed by the fact that females started building their nests more frequently where other females of their species were simultaneously digging. The distances between subsequent nests dug by individual females were shorter than those obtained by random simulations. However, this pattern seemed to depend on the tendency to dig close to conspecifics rather than remain in the vicinity of previous nests, suggesting that females' experience matters to future decisions only on a large scale. Nesting patches within nest aggregations largely overlapped between species, but the nests of each species were generally not closer to heterospecific nests than expected by chance, suggesting that females are neither repulsed by, nor attracted to, congenerics within nest aggregations. A role of the spatial distribution of natural enemies on the observed nesting patterns seemed unlikely. Bembix digger wasp nest aggregations seem thus to be primarily the result of female–female attraction during nest‐settlement decisions, in accordance with the ‘copying’ mechanisms suggested for nesting vertebrates.     

Behavioral responses of the parasitoid Melittobia digitata to volatiles emitted by its natural and laboratory hosts

Responses of macropterous females of the ectoparasitoid Melittobia digitata Dahms (Hymenoptera: Eulophidae) to direct and indirect cues emitted by its natural hosts as well as laboratory hosts were investigated using a Y‐tube olfactometer. To locate the nest of mud dauber wasps, Trypoxylon politum Say (Hymenoptera: Crabronidae), and one of their inquilines, Anthrax spec., parasitoids exploit volatiles from the freshly built nest mud and the empty cocoon constructed by the wasps, as well as their meconium. However, the parasitoids did not respond to odors emitted by older nest mud or by the host stages that are attacked (T. politum prepupae and Anthrax spec. larvae). Melittobia digitata was not attracted to direct volatiles released by the dipteran hosts Anastrepha ludens Loew (Diptera: Tephritidae) (a natural host) and Sarcophaga bullata (Parker) (Diptera: Sarcophagidae) (a laboratory host). Based on our results, we suggest that M. digitata adopts a ‘sit and wait’ strategy to locate mud dauber wasps, relying mainly on indirect host‐related cues: females search for nests that are under construction and once found, they wait inside the cell until the host completes its cocoon and releases meconium, an indicator that is associated with host suitability. No attraction was found to dipteran hosts, suggesting that parasitization of these hosts may be incidental, due to the broad host plasticity of Melittobia wasps.     

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.     

Does Conspecific Attraction Affect Nest Choice in Budgerigars (Melopsittacus undulatus: Psittacidae: Aves)?

In their native Australia the budgerigar's breeding season is very short and rapid nest establishment is crucial. To minimize the time required to locate and establish nesting sites, female budgerigars were predicted to use cues on nest site locations provided by other females in their colony. Experimental results from captive birds indicated that, while females can be attracted to nests that are occupied by other females, the social and breeding experience of individuals affects this use of conspecific cues. Females that have had altercations with other females that resulted in physical injuries showed an aversion to apparently occupied nestboxes. Females that had no such social experience prior to testing were attracted to apparently occupied nestboxes. Breeding experience, as well as social experience, may have an effect on the use of conspecifics as cues. Experienced breeding females were less likely to be attracted to apparently occupied nestboxes than were females that had never bred before.     

Nest preference and ecology of cavity-nesting bees (Hymenoptera: Apoidea) in Punjab,Pakistan

Among bees, 85 % are solitary species, most of them are ground-nester and some are cavity-nesting and construct their nests in pre-existing cavities. This work was conducted to evaluate the substrate preference and nest architecture (acceptance, occupation percentage, seasonality and parasitism) of cavity-nesting bees in different substrates. Trap nests offered comprised five different materials (drilled cavities in wood, bamboo, cardboard tubes, plastic soda straws, and mud blocks). These were installed in four districts for two years. The nesting cavities of five different diameters (6, 8, 10, 12 and 14 mm) were provided in each nesting material with an average length of 180 ± 9.92 mm. In all the materials, 5400 nesting cavities were offered, out of which 628 were colonized by bees from two families (Megachilidae and Apidae) and six species (Megachile cephalotes, M. lanata, M. bicolor, Xylocopa basalis, X. fenestrata and Ceratina smaragdula), including one parasitic bee (Euaspis carbonaria). The bee species differed significantly in occupying five nesting materials. The most preferred diameters were 8 mm and 10 mm, with 52.20% and 29.45% of colonization, respectively. Nesting was done throughout the year except in winter. This study will serve as a baseline for future studies and conservation programs of cavity-nesting bees in Pakistan.     

Effect of the entomopathogenic fungus,Entomophthora muscae (Zygomycetes: Entomophthoraceae), on sex pheromone and other cuticular hydrocarbons of the house fly,Musca domestica

House fly (Musca domestica) males are highly attracted to dead female flies infected with the entomopathogenic fungus Entomophthora muscae. Because males orient to the larger abdomen of infected flies, both visual and chemical cues may be responsible for the heightened attraction to infected flies. Our behavioral assays demonstrated that the attraction is sex-specific-males were attracted more to infected females than to infected males, regardless of cadaver size. We examined the effect of E. muscae on the main component of the house fly sex pheromone, (Z)-9-tricosene, and other cuticular hydrocarbons including n-tricosane, n-pentacosane, (Z)-9-heptacosene, and total hydrocarbons of young (7 days old) and old (18 days old) virgin females. Young E. muscae-infected female flies accumulated significantly less sex pheromone and other hydrocarbons on their cuticular surface than uninfected females, whereas the cuticular hydrocarbons of older flies were unaffected by fungus infection. These results suggest that chemical cues other than (Z)-9-tricosene, visual cues other than abdomen size, or a combination of both sets of cues might be responsible for attraction of house fly males to E. muscae-infected females.     

An overview of proximate factors affecting the nesting behavior of solitary wasps and bees (Hymenoptera: Aculeata) in preexisting cavities in wood

Guilds of Aculeate solitary wasps and bees that nest in preexisting cavities in wood are important components of terrestrial ecosystems because they engage in several ecological interactions (e.g. predation and pollination) with other species of plants and animals. Spatial and temporal variations in richness and abundance of solitary wasps and bees can be related to changes in environmental structure and in the diversity of other groups of organisms. The nesting period of these Aculeata is their most critical life cycle stage. Females of solitary wasp and bee species invest relatively more time constructing and provisioning their nests than do females of social species. Differently from species that nest in the soil or construct exposed nests, the main factors affecting the reproductive success of solitary species nesting in preexisting wood holes are still unknown. Our objective is to provide an overview of the role of proximate causes of nesting failure or success among solitary wasps and bees (Aculeata), for designing effective conservation and management strategies for these Hymenoptera.     

Predation Cues in Solitary bee Nests

Predation at the nesting site can significantly affect solitary bees’ reproductive success. We tested female red mason bees’ (Osmia bicornis L.) acceptance of potential nesting sites, some of which were marked with cues coming from predated conspecifics (crushed bees) or from a predator itself (rodent excreta). In our experiment, females did not avoid nests marked with either of the two predator cues. We suggest that bee females do not recognize these two cues as risky. Alternatively, costs of abandoning natal aggregation might be too high compared with any perceived predation risk of staying. Moreover, the presence of crushed bees can provide positive information about the presence of conspecifics and, possibly, information about a nesting aggregation that may be preferred by bees when choosing a nesting site.     

Effects of Fungicide and Adjuvant Sprays on Nesting Behavior in Two Managed Solitary Bees,Osmia lignaria and Megachile rotundata

There is a growing body of empirical evidence showing that wild and managed bees are negatively impacted by various pesticides that are applied in agroecosystems around the world. The lethal and sublethal effects of two widely used fungicides and one adjuvant were assessed in cage studies in California on blue orchard bees, Osmia lignaria, and in cage studies in Utah on alfalfa leafcutting bees, Megachile rotundata. The fungicides tested were Rovral 4F (iprodione) and Pristine (mixture of pyraclostrobin + boscalid), and the adjuvant tested was N-90, a non-ionic wetting agent (90% polyethoxylated nonylphenol) added to certain tank mixtures of fungicides to improve the distribution and contact of sprays to plants. In separate trials, we erected screened cages and released 20 paint-marked females plus 30–50 males per cage to document the behavior of nesting bees under treated and control conditions. For all females in each cage, we recorded pollen-collecting trip times, nest substrate-collecting trip times (i.e., mud for O. lignaria and cut leaf pieces for M. rotundata), cell production rate, and the number of attempts each female made to enter her own or to enter other nest entrances upon returning from a foraging trip. No lethal effects of treatments were observed on adults, nor were there effects on time spent foraging for pollen and nest substrates and on cell production rate. However, Rovral 4F, Pristine, and N-90 disrupted the nest recognition abilities of O. lignaria females. Pristine, N-90, and Pristine + N-90 disrupted nest recognition ability of M. rotundata females. Electroantennogram responses of antennae of O. lignaria females maintained in the laboratory did not differ significantly between the fungicide-exposed and control bees. Our results provide the first empirical evidence that two commonly used fungicides and a non-ionic adjuvant can disrupt nest recognition in two managed solitary bee species.     

Not all who wander are lost: nest fidelity in <Emphasis Type="Italic">Xylocopa virginica</Emphasis> examined by mark recapture

Relocation to novel nests (sometimes called drifting) in flying Hymenoptera is often interpreted as the result of navigation error and guard bees erroneously admitting foreign individuals into the nest. We studied nest fidelity and nest relocation of both females and males in a nesting aggregation of Xylocopa virginica in southern Ontario, Canada, where females can nest either solitarily or socially. Adult female and male bees were trapped at nest entrances, individually paint marked, and then released. Subsequent recapture patterns were used to assess nest fidelity: that is, how faithful individuals were to their home nest and how often they moved to another nest. Bees were considered to have relocated if they were recaptured in a nest different from the one in which they were initially trapped, indicating that they had spent at least one night in a new nest. Some females were only captured in one nest, some occasionally moved to new nests, temporarily or permanently, and a few were never caught in the same nest twice. In addition, females relocated to nests that were further away in 2007 when population density was low, suggesting that they seek out and claim nesting spaces when they are available. Males relocated more frequently than females, with most drifting from nest to nest in no obvious pattern. This indicates that males spend the night wherever space is available or in nests nearest to their territories. This study reveals that for both female and male X. virginica, nest membership is not as stable as once thought.     

Chalkbrood transmission in the alfalfa leafcutting bee: the impact of disinfecting bee cocoons in loose cell management systems

Understanding pathogen transmission could illuminate new methods for disease prevention. A case in point is chalkbrood in the alfalfa leafcutting bee [Megachile rotundata (F.)]. Propagation of this solitary bee is severely hampered by chalkbrood, a larval disease caused by Ascosphaera aggregata (Ascomycota). Alfalfa leafcutting bees nest in existing cavities in wood or hollow reeds and overwinter as larvae. In the early summer, emerging adults frequently must chew through dead, diseased siblings that block their exit, becoming contaminated with chalkbrood spores in the process. When alfalfa leafcutting bees are used as a commercial pollinator, the cocoons are removed from nesting boards to reduce chalkbrood transmission, but the disease is still common. To determine if these removed cocoons (called loose cells) are an important source of disease transmission, they were disinfected with a fungicide before bees were incubated, and released in the field. Chalkbrood prevalence among the progeny of the treated bees was reduced up to 50% in one field trial, but not significantly when tested in an on-farm trial. Thus, substantial disease transmission still occurred when the loose cells were disinfected, and even when clean nesting materials were used. In conclusion, pathogen transmission must still be occurring from another source that has yet to be identified. Another possible source of transmission could arise from bees that emerge midsummer in populations with a high percent of multivoltinism, but dirty nesting boards and feral bees also may be minor sources of transmission.     

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.     

Bee Tracker—an open‐source machine learning‐based video analysis software for the assessment of nesting and foraging performance of cavity‐nesting solitary bees

The foraging and nesting performance of bees can provide important information on bee health and is of interest for risk and impact assessment of environmental stressors. While radiofrequency identification (RFID) technology is an efficient tool increasingly used for the collection of behavioral data in social bee species such as honeybees, behavioral studies on solitary bees still largely depend on direct observations, which is very time‐consuming. Here, we present a novel automated methodological approach of individually and simultaneously tracking and analyzing foraging and nesting behavior of numerous cavity‐nesting solitary bees. The approach consists of monitoring nesting units by video recording and automated analysis of videos by machine learning‐based software. This Bee Tracker software consists of four trained deep learning networks to detect bees that enter or leave their nest and to recognize individual IDs on the bees’ thorax and the IDs of their nests according to their positions in the nesting unit. The software is able to identify each nest of each individual nesting bee, which permits to measure individual‐based measures of reproductive success. Moreover, the software quantifies the number of cavities a female enters until it finds its nest as a proxy of nest recognition, and it provides information on the number and duration of foraging trips. By training the software on 8 videos recording 24 nesting females per video, the software achieved a precision of 96% correct measurements of these parameters. The software could be adapted to various experimental setups by training it according to a set of videos. The presented method allows to efficiently collect large amounts of data on cavity‐nesting solitary bee species and represents a promising new tool for the monitoring and assessment of behavior and reproductive success under laboratory, semi‐field, and field conditions.     

Solitary and group nesting in the orchid bee <Emphasis Type="Italic">Euglossa hyacinthina</Emphasis> (Hymenoptera,Apidae)

Summary Orchid bees (Euglossini) provide a potentially informative contrast for examining origins of advanced social behaviour in bees because they are the only tribe in the apine clade that do not form large colonies or have queens and workers. We investigated natural nests of Euglossa hyacinthina Dressler, an orchid bee that nests singly or in groups. By comparing the two types of nests, we examined if individuals in a group merely share the nest (are communal) or exhibit a level of social organization where there is reproductive division of labour among the females. Observations are consistent with communal nesting, indicating that all females in group nests are reproductively similar to the solitary nesting females because the provisioning of young, as well as the ovary development and mating status of females sharing nests were not different than that of solitary-nesting females. Also, multiple female nests did not produce a female-biased brood as predicted for nests with reproductive division of labour. We also investigated potential advantages of group nesting vs. individual nesting. We demonstrate that per capita offspring production is lower in nests with more than one female. However, we found that nests with single females were left unattended for longer periods of time during foraging, and that there was a high incidence of natural enemy attack in nests when females were absent. Group and solitary nesting may be advantageous under different conditions.Received 3 December 2002; revised 7 March 2003; accepted 2 April 2003.     

Evolution of nesting behaviour and kleptoparasitism in a selected group of osmiine bees (Hymenoptera: Megachilidae)

The construction of nests to rear offspring is restricted to vertebrates and few insect taxa, such as termites, wasps, and bees. Among bees, species of the family Megachilidae are characterized by a particularly high diversity in nest construction behaviour. Many megachilid bees nest in excavated burrows in the ground, others place their brood cells in a variety of above‐ground cavities or attach them to the surface of a substrate, and yet others have adopted a kleptoparasitic habit. Evolutionary transitions between the different nesting sites and between conventional nesting and kleptoparasitism in bees are poorly understood. In the present study, we traced the evolution of nesting site selection and kleptoparasitism in the Annosmia–Hoplitis group (Osmiini), which displays an exceptionally high diversity in nesting behaviour. We found that the evolution of nesting behaviour proceeded unidirectionally from nesting in excavated burrows in the ground to nesting in rock depressions and cavities, followed by the colonization of snail shells and insect borings in dead wood or hollow stems. Kleptoparasitism evolved once and the kleptoparasitic species have derived from the same lineage as their hosts. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 108 , 349–360.