Description and validation of an improved method to feed solitary bees (Osmia spp.) known amounts of pesticides.

Pesticide exposure is an important driver of bee declines. Laboratory toxicity tests provide baseline information on the potential effects of pesticides on bees, but current risk assessment schemes rely on one species, the highly social honey bee, Apis mellifera, and there is uncertainty regarding the extent to which this species is a suitable surrogate for other pollinators. For this reason, Osmia cornuta and Osmia bicornis have been proposed as model solitary bee species in the EU risk assessment scheme. The use of solitary bees in risk assessment requires the development of new methodologies adjusted to the biology of these species. For example, oral dosing methods used with honey bees cannot be readily applied to solitary bees due to differences in feeding behaviour and social interactions. In this study, we describe the "petal method", a laboratory feeding method, and validate its use in acute and chronic exposure oral tests with Osmia spp. We conducted five experiments in which we compared the performance of several artificial flowers combining visual and olfactory cues against the petal method, or in which variations of the petal method were confronted. We then use the results of these experiments to optimize the feeding arenas and propose standardized methods for both acute and chronic exposure tests. The petal method provides high levels of feeding success, thus reducing the number of bees needed. It works with a wide variety of petal species and with both female and male Osmia spp., thus ensuring reproducibility across studies. To validate the use of the petal method in ecotoxicology tests, we assess the toxicity of a standard reference insecticide, dimethoate, in O. cornuta adults and determine LD50 values for this species. The petal method should facilitate the inclusion of solitary bees in risk assessment schemes therefore increasing the protection coverage of pesticide regulation.

Pesticide Contamination in Native North American Crops, Part II-Comparison of Flower, Honey Bee Workers, and Native Bee Residues in Lowbush Blueberry.

In lowbush blueberry fields, we conducted residue analysis comparing flowers, trapped pollen (honey bee and Osmia spp.), and collected bees (honey bee workers, bumble bee queens, and non-Bombus spp. wild native bees). The study was conducted from 2012 to 2014. The number of pesticide residues, total concentrations, and risk to honey bees (Risk Quotient) on flowers were not significantly different from those determined for trapped honey bee pollen (except in one study year when residues detected in flower samples were significantly lower than residue numbers detected in trapped pollen). The compositions of residues were similar on flowers and trapped pollen. The number of residues detected in honey bee pollen was significantly greater than the number detected in Osmia spp. pollen, while the total concentration of residue was not different between the two types of pollen. The risk to honey bees was higher in trapped honey bee pollen than in trapped Osmia spp. pollen. The analysis of honey bee workers, native bumble bee queens, and native solitary bees showed that although more pesticide residues were detected on honey bee workers, there were no differences among the bee taxa in total residue concentrations or risk (as estimated in terms of risk to honey bees).

More is less: mass-flowering fruit tree crops dilute parasite transmission between bees.

Parasites influence wild bee population dynamics and are regarded as one of the main drivers of wild bee decline. Most of these parasites are mainly transmitted between bee species via the use of shared floral resources. Disturbance of the plant-pollinator network at a location can hence disturb the transmission of these parasites. Expansion and intensification of agriculture, another major driver of wild bee decline, often disturbs local plant-pollinator networks by altering the availability and diversity of floral resources. Mass-flowering crops are an extreme example as they provide an abundance of floral resources for a short period of time, substantially altering the present plant-pollinator network. This likely has repercussions on parasite transmission in the pollinator community. Using the bloom of mass-flowering crops we tested the hypothesis that an increase in floral resources can dilute parasite transmission in the pollinator community. To test this, we analysed the presence of parasites in the pollen of the brood cell provisions of Osmia spp., collected from trap nests placed in apple and sweet cherry orchards. We collected pollen at several time intervals during and after mass bloom, and found that pollen collected during mass bloom had significantly lower parasite prevalence compared with pollen collected after mass bloom. Furthermore, using pollen barcoding data we found that the presence of MFCs in pollen was a good predictor for lower parasite prevalence. Taken together, our results indicate that an increase in flower availability can reduce parasite transmission between bees.

Towards Integrated Pest and Pollinator Management in Intensive Pear Cultivation: A Case Study from Belgium.

Recently, the concept of Integrated Pest Management (IPM) was further extended into Integrated Pest and Pollinator Management (IPPM). Implementation of IPPM strategies entails the combination of actions for pest and pollinator management providing complementary or synergistic benefits for yield and/or quality of the harvest. The aim of this study was to examine IPPM elements (i.e., mixed hedgerow, nesting boxes for mason bees, Osmia spp.) and demonstrate their impact in the practical context of modern commercial fruit cultivation in a 4-year case study in an intensive 'Conference' pear orchard. The outcomes of visual observations during transect walks and molecular analysis of pollen collected by mason bees, showed the importance of additional floral resources for the presence of mason bees and other pollinating insects in the orchard environment. Pear quality assessments indicated that insect-mediated pollination had a significant positive impact, with a tendency for higher quality pears in the close vicinity of Osmia nesting boxes. However, despite the fact that pear pollen was also detected in Osmia spp. nest cells, the amount and frequency of pear pollen collection for their nest built-up turned out to be rather low. In the same intensive pear orchard studied for pollination effects, we simultaneously demonstrate the impact of a mixed hedgerow to enhance integrated pest control.

Bifidobacterium xylocopae sp. nov. and Bifidobacterium aemilianum sp. nov., from the carpenter bee (Xylocopa violacea) digestive tract.

Social bees harbor a community of gut mutualistic bacteria, among which bifidobacteria occupy an important niche. Recently, four novel species have been isolated from guts of different bumblebees, thus allowing to suppose that a core bifidobacterial population may be present in wild solitary bees. To date there is sparse information about bifidobacteria in solitary bees such as Xylocopa and Osmia spp., this study is therefore focused on the isolation and characterization of bifidobacterial strains from solitary bees, in particular carpenter bee (Xylocopa violacea), builder bee (Osmia cornuta), and red mason bee (Osmia rufa). Among the isolates from Osmia spp. no new species have been detected whereas among Xylocopa isolates four strains (XV2, XV4, XV10, XV16) belonging to putative new species were found. Isolated strains are Gram-positive, lactate- and acetate-producing and possess the fructose-6-phosphate phosphoketolase enzyme. Full genome sequencing and genome annotation were performed for XV2 and XV10. Phylogenetic relationships were determined using partial and complete 16S rRNA sequences and hsp60 restriction analysis that confirmed the belonging of the new strains to Bifidobacterium genus and the relatedness of the strains XV2 and XV10 with XV16 and XV4, respectively. Phenotypic tests were performed for the proposed type strains, reference strains and their closest neighbor in the phylogenetic tree. The results support the proposal of two novel species Bifidobacterium xylocopae sp. nov. whose type strain is XV2 (=DSM 104955T=LMG 30142T), reference strain XV16 and Bifidobacterium aemilianum sp. nov. whose type strain is XV10 (=DSM 104956T=LMG 30143T), reference strain XV4.