Insects

Apis cerana cerana is a subspecies of eastern honeybee, Apis cerana. Nosema ceranae is a widespread fungal parasite of honeybee, causing heavy losses for beekeeping industry all over the world. In this article, total RNA of normal midguts (AcCK1, AcCK2) and N. ceranae-infected midguts of A. c. cerana workers at 7 d and 10 d post inoculation (AcT1, AcT2) were respectively isolated followed by strand-specific cDNA library construction and next-generation RNA sequencing. In tolal, 56270223688, 44860946964, 78991623806, and 92712308296 raw reads were derived from AcCK1, AcCK2, AcT1 and AcT2, respectively. Following strict quality control, 54495191388, 43570608753, 76708161525, and 89467858351 clean reads were obtained, with Q30 value of 95.80%, 95.99%, 96.07% and 96.04%, and GC content of 44.20%, 43.44%, 44.83% and 43.63%, respectively. The raw data were submitted to the NCBI Sequence Read Archive database and connected to BioProject PRJNA562784. These data offers a valuable resource for deep investigation of mechanisms underlying eastern honeybee responding to N. ceranae infection and host-fungal parasite interaction during microsporidiosis. Value of the DataO_LICurrent dataset offers a valuable resource for exploring mRNAs, lncRNAs and circRNAs involved in response of A. c. cerana worker to N. ceranae infection. C_LIO_LIThe accessible data can be used to investigate differential expression pattern and regulatory network of non-coding RNAs in A. c. cerana workers midguts responding to N. ceranae challenge. C_LIO_LIThis data will enable a better understanding of the molecular mechanism regulating eastern honeybee-N. ceranae interaction. C_LI

Apis mellifera ligustica is a subspecies of western honeybee, Apis mellifera. Nosema ceranae is known to cause bee microspodiosis, which seriously affects bee survival and colony productivity. In this article, Nanopore long-read sequencing was used to sequence N. ceranae-infected and un-infected midguts of A. m. ligustica workers at 7 d and 10 d post inoculation (dpi). In total, 5942745, 6664923, 7100161 and 6506665 raw reads were respectively yielded from AmT1, AmT2, AmCK1 and AmCK2, with average lengths of 1148, 1196, 1178 and 1201 bp, and N50 of 1328, 1394, 1347 and 1388 bp. The length distribution of raw reads from AmT1, AmT2, AmCK1 and AmCK2 was ranged from 1 kb to more than 10 kb. Additionally, the distribution of quality score of raw reads from AmT1 and AmT2 was among Q6[~]Q12, while that from AmCK1 and AmCK2 was among Q6[~]Q16. Further, 5745048, 6416987, 6928170, 6353066 clean reads were respectively gained from AmT1, AmT2, AmCK1 and AmCK2, and among them 4172542, 4638289, 5068270 and 4857960 were identified as being full-length. After removing redundant reads, the length distribution of remaining full-length transcripts was among 1 kb[~]8 kb, with the most abundant length of 2 kb. The long-read transcriptome data reported here contributes to a deeper understanding of the molecular regulating N. ceranae-response of A. m. ligustica and host-fungal parasite interaction during microsporidiosis.

Apis cerana cerana is a subspecies of eastern honeybee, Apis cerana. Ascosphaera apis is a widespread fungal pathogen of honeybee, leading to chalkbrood, which results in heavy losses for beekeeping industry. In this article, 4-, 5-, and 6-day-old larval guts of un-infected (AcCK1, AcCK2, and AcCK3) and A. apis-infected A. c. cerana (AcT1, AcT2, and AcT3) were sequenced by next generation sequencing. Totally, 73830148, 96586212, 94552744, 76672564, 90954858, and 83418832 raw reads were respectively produced from AcCK1, AcCK2, AcCK3, AcT1, AcT2, and AcT3. The sequencing depth was enough to detect all expressed genes. After strict quality control, 73775592, 96513798, 94495000, 76593924, 90870608 and 83339288 clean reads were obtained, with a mean GC content of 48.54%. Additionally, average Q20 and Q30 for aforementioned six groups were 98.10% and 94.36%, respectively. Moreover, 45302685, 65872823, 52709987, 49947838, 56476339, and 42657156 clean reads from above-mentioned six groups were mapped to the reference genome of Apis cerana, respectively. In addition, exons were the most abundant regions in reference genome mapped by clean reads, followed by intergenic regions and introns. Our data presented here can be used to identify long non-coding RNAs (lncRNAs), circlular RNAs (circRNAs), mRNAs and their regulatory networks engaged in response of eastern honeybee larvae to A. apis infection, and decipher molecular mechanisms underlying host-pathogen interaction. Value of the DataO_LIThe current data can be used for exploration of mRNAs, lncRNAs, circRNAs and their competing endogenous RNA (ceRNA) networks engaged in response of A. c. cerana larvae to A. apis infection. C_LIO_LIThis data provides a valuable genetic resource for better understanding non-coding RNA-mediated cross-kingdom regulation of eastern honey larvae to A. apis. C_LIO_LIThe accessible data offers novel insights into understanding the molecular mechanism underlying interaction between eastern honeybee larvae and A. apis. C_LI

Comprehensive expression datasets were constructed for Apis mellifera and Apis cerana japonica. Post-oviposition day 6 to day 58 samples of A. mellifera workers (larva to adult); day 9, 10, 12, and 13 samples of A. mellifera queen (larva to pupa); and day 9 to day 18 samples of A. cerana japonica workers (larva to adult) were prepared, and RNA-Seq data were obtained. For A. cerana japonica, reference transcript sequence data, predicted amino acid sequence data, and functional annotation data were generated based on the genome sequence and RNA-Seq data. Using the transcript sequence and RNA-Seq data, comprehensive expression data for all transcripts of A. mellifera and A. cerana japonica were prepared. Hierarchical clustering analyses and the used sample preparation method ensured that both sets of expression data were reliable for use as comprehensive reference expression datasets. Therefore, these data are applicable for honey bee research or for comparative or evolutionary studies on insect species or social insect species at the genetic and molecular levels.

Neonicotinoids, including imidacloprid, harm Apis mellifera in a number of ways, among others by impairing body maintenance and immunity. Energy resources are important to prevent this, particularly, as we hypothesized, in the fat body tissue. But hormesis (adaptative, diaphasic response to a stressor) was also reported in the energy-dependent traits of bees exposed to sublethal doses of imidacloprid. Consequently, concentrations/activities of respiratory and citric cycle compounds (Acetyl-CoA, IMH-2, AKG, succinate, fumarate, NADH2, CoC, COX, and ATP) were compared in the hemolymph and fat bodies of honeybees fed with diets containing 200 ppb (IM-200), 5 ppb (IM-5; sublethal), and 0 ppb of ID. The values of the compounds were higher in the fat body than in the hemolymph, where the variability was higher in hemolymph. The pattern of response to ID was the same in both tissues, but differed between IM-200 and IM-5. The concentrations of the strongly correlated NADH2, ATP and acetyl-CoA decreased both in ID-200 and ID-5, whereas the levels of the remaining compounds decreased in ID-200 but increased in ID-5. Decreased ATP levels in each diet show that the pesticide impairs the fat-body energy metabolism in spite of hormesis in six of the nine respiratory and citric cycle compounds even in the low, residual doses. We believe that sublethal doses of imidacloprid decrease energy demands, as they decreased the levels of ATP, and acetyl-CoA.

Nosema ceranae is a widespread fungal parasite that infects both adult honeybee and honeybee larvae, leading to microsporidiosis, which seriously affects bee health and apicultural industry. In this article, genome sequencing of clean spores of N. ceranae was conducted using third-generation Pacific Biosciences (PacBio) single molecule real time (SMRT) sequencing technology. In total, 152671 subreads were obtained after quality control of raw reads from PacBio SMRT sequencing, with a N50 and average length of 14422 bp and 11310 bp, respectively. Additionally, the length distribution of subreads was from 10000 bp to more than 50000 bp. Nineteen scaffords with a total length of 7354221 bp were assembled, and the N50, N90 and maximum scafford length were 728543 bp, 198795 bp and 1917792 bp, respectively. The GC content was 25.97%. Furthermore, by integration of genes predicted from de novo and homology-based methods, 3112 N. ceranae genes were finally assembled, with a total length of 2730179 bp and mean length of 877.31 bp. In addition, the total length and mean length of exons were 2657637 bp and 854 bp, respectively; and the total length and mean length of introns were 72542 bp and 23.31 bp, respectively. The genome sequencing data documented here will give deep insights into the molecular biology of N. ceranae, facilitate exploration of genes and pathways associated with toxin factors and infection-related factors, and benefit research on comparative genomics and phylogenetic diversity of Nosema species.

Odorant binding proteins (OBPs) in insects bind to volatile chemical cue and help in their binding to odorant receptors. The odor coding hypothesis states that OBPs may bind with specificity to certain volatiles and aid the insect in various behaviours. Honeybees are eusocial insects with complex behaviour that requires olfactory inputs. Here, we have identified and annotated odorant binding proteins from the genome of the dwarf honey bee, Apis florea using an exhaustive homology-based bioinformatic pipeline and analyzed the evolutionary relationships between the OBP subfamilies. Our study suggests that Minus-C subfamily may have diverged from the Classic subfamily of odorant binding proteins in insects.

Oxidative stress (OX) is a state of imbalance between antioxidants and reactive oxygen species, which are the byproducts of oxidative phosphorylation in the mitochondria. Different landscapes: conventional, organic, and roadside use different pesticides and pest control management practices pest control. The roadside landscape is a control for minimum application of herbicides and management. Two important pollinators, honey bees (Apis mellifera) and small carpenter bees (Ceratina calcarata) are studied here for their abiotic stress from different landscape. It is unknown whether the differences in these landscapes affect the oxidative stress of these two pollinators. Conventional farms offer the most exposure to pesticides, which have been related to increased stress levels. We hypothesized that honey bees and small carpenter bees (SCB) from conventional farms would experience the highest levels of OX. Results from the lipid assay indicated that honey bees and SCB of organic and roadside landscapes experienced the lowest and highest OX levels. The results of the protein carbonyl assay indicated a significant difference found between conventional and organic bees (p < 0.05), based on Tukeys post-hoc test. Previous research has shown that feral bees have a tolerance to high stress levels. The same could be true of honey bees and SCB from roadside landscapes, where the conditions imitate natural habitats. Organic farms use naturally derived pesticides and moderate management practices, likely contributing to their lower levels. Examining how these pollinators are affected by different farm landscapes can reveal the benefits and detriments of their corresponding management practices. This will lead to streamlined practices that are healthier for pollinators.

Feeding experiments are standard tools in the pollinator risk assessment. The design (Oomen et al 1992) was developed to test insect growth regulators and herbicides. In recent years there was an update (Luckmann & Schmitzer 2015) on the outline in order to also focus on the advantage of different rates making a dose response design possible where exposure levels are known. Additionally, this design gives the possibility to test different rates for honey bee colonies foraging in the same landscape. The main objective of the experiment presented here was to determine the natural variability of foragers losses of hives fed with a sub-lethal neonicotinoid concentration compared to an untreated control. Other objectives were to see if the neurotoxic exposure results in any observable sub-lethal effects and to find out if losses can be correlated to hive development. This was assessed with traditional methods and a novel, visual monitoring device.

A parasitic mite of honey bee brood (Tropilaelaps mercedesae), is spreading through populations of Apis mellifera honey bees in new regions and poses a major threat to honey bee health. Despite its clear threat, the biology of this mite is poorly understood, with gaps on such fundamental issues as how fast its populations can grow. This leaves the beekeeping world underprepared to plan for its arrival and management. In this study, we documented the growth of T. mercedesae populations in untreated A. mellifera colonies in Thailand and South Korea, and did the same for another parasitic mite (Varroa destructor) when possible. We found that the population growth of T. mercedesae was variable but could reach high levels (daily r of 0.010, 0.036, and 0.057), while the population growth of V. destructor (r = 0.021) matched previous estimates. Our results indicate that T. mercedesae populations can grow rapidly but they do not always attain this potential. Based on our results, humidity should be studied as a potential driver of population growth. If future work can reveal key drivers of T. mercedesae population growth, this would help predict infestations and help design management strategies that exploit the pest's biological vulnerabilities.

The western honey bee, Apis mellifera, faces a new threat from the spread of parasitic Tropilaelaps (Tropi) mites, specifically T. mercedesae, which adds additional complexity to an apicultural landscape heavily impacted by Varroa destructor. In this study conducted in central Thailand, we investigated the efficacy of two methods of applying formic acid and a thermal remediation technique in controlling Tropi mites and Varroa, focusing our attention on the reproductive stage of the mites which is restricted to capped brood cells. Results revealed that both formic acid treatments (Formic Pro and liquid formic acid) demonstrated an immediate and substantial reduction in live Tropi and Varroa populations, maintaining near-zero levels for the 3- week duration of the study. In contrast, thermal remediation, employing heating pads, exhibited a more gradual decline, achieving an 85.42% reduction in Tropi mites and a 92.33% reduction in Varroa mites by week three. Notably, heat-treated colonies experienced an unexpected resurgence in mite populations during week two. The findings contribute valuable insights into potential strategies for mitigating the threat of Tropi mites and highlight the urgency of further research to safeguard global honey bee populations.

In the era of global warming, much attention has been paid to the possibility that high temperature may influence diverse insects not only directly but also indirectly via effects on their symbiotic microorganisms. The stinkbug Plautia stali develops a midgut symbiotic organ that harbors a specific bacterial symbiont indispensable for its growth and survival. Being maintainable in laboratory and tractable experimentally, P. stali is recently highlighted as a model system to investigate the mechanisms underpinning insect-microbe symbiotic interactions. In this study, we reared newly-emerged adult insects of P. stali under different temperature conditions for 8 days and monitored how their symbiotic organs and symbiotic bacteria are affected. While all insects survived at temperatures from 25{degrees}C to 37{degrees}C, some insects died at 38{degrees}C, 39{degrees}C and 40{degrees}C, wherein mortality rates increased as temperature elevated. While the symbiotic organs of the normal insects exhibited vivid yellow color, the symbiotic organs of the insects reared at 35{degrees}C or higher frequently exhibited abnormal colors such as pale yellow, yellowish white, or white, the extent of which became more severe as temperature elevated. Symbiont quantification revealed that, while the symbiont titers were almost constant for 8 days at 25{degrees}C and 30{degrees}C, the symbiont titers on the 8th day drastically declined to 1/100 at 35 and 1/10000 at 37{degrees}C and 39{degrees}C. Based on these results, we propose that rearing at 37{degrees}C for a week is a recommended treatment regime by which the symbiont is effectively suppressed with minimal damage to the host insect.

Fipronil, a non-competitive antagonist of {gamma}-aminobutyric acid receptors (GABA-Rs), inhibits the flow of chloride ions in the nerve cells, leading to the insect death. In a previous work, the volatolome of bees chronically exposed to this insecticide (0.5 and 1 g/L) was analyzed and characterize the Apis mellifera metabolic responses. Two Volatile Organic Compounds or VOCs (1-octen-3-ol and 2,6-dimethylcyclohexanol) produced by bee were highlighted. We assumed that these VOCs could act as modulators of the GABA-R to counteract the effect of fipronil. The toxicity of these VOCs was tested on emerging bees for concentrations ranging from 0 to 3.4 g/L. Survival and the sucrose consumption were recorded during 21 days of chronic exposure. 1-octen-3-ol and 2,6-dimethylcyclohexanol did not significantly affect the proportion of survival at the end of the experiment, whatever their concentration. 1-octen-3-ol (0.5, 1, 1.7 and 3.4 g/L) was chosen to assess its effect in the case of co-exposure with fipronil (0.5 and 1 g/L). A beneficial effect on survival at 21 days was observed with an average improvement in survival rate for the co-exposed groups. This positive effect was not related to the VOC concentration and could be a direct effect (interaction with GABA-R) or a hormetic effect (global improvement of bee fitness).

O_LIHemiptera and Thysanoptera insects pose persistent threats to agricultural production. Conventional management strategies involve the release of chemical or plastic agents, causing adverse environmental and global health issues. Notably, Nicotiana benthamiana, a globally utilized model plant, exhibits remarkable lethal effects and attraction towards these pests. C_LIO_LIIn this study, we explored the potential of using N. benthamiana for Hemiptera and Thysanoptera pest control in the laboratory and field. Through net cover and three field assays over two years, we demonstrated the efficacy and benefits of using N. benthamiana as a field-deployed pest control dead-end trap. C_LIO_LIN. benthamiana demonstrated nearly 100% lethality to whiteflies, aphids, and thrips, with emitted volatiles attracting these insects. Field trials showed that potted and planted N. benthamiana blocks and traps whiteflies and thrips from several Solanaceae and Cucurbitaceae crops effectively, comparable to common commercial yellow and blue sticky boards. Moreover, N. benthamiana in the field exhibits robust growth in commercial greenhouses without negatively impacting crop growth, natural enemies, and pollinators. C_LIO_LIOur study introduces an innovative, easily implementable, and sustainable approach for controlling Hemiptera and Thysanoptera pests. Moreover, it unveils the novel utility of N. benthamiana in field-based pest management. C_LI

Immunosuppression and premature senescence are main risks of exposure to toxic compounds that might define individual longevity and the fate of natural animal populations. However, these sublethal effects have not been studied in insects that are of fundamental importance for human economy and well-being such as dung beetles. We exposed adult dung beetles Euoniticellus intermedius to ivermectin, a commonly used antiparasitic drug in cattle that is excreted in dung, and measured immune activity through phenoloxidase (PO) and its zymogen, prophenoloxidase (proPO), in males and females of different ages. We predicted that both ivermectin exposure and age would reduce immune activity, with the negative effect of ivermectin being more pronounced in older individuals, revealing immunosenescence. Despite PO and proPO activities decreased with age and ivermectin exposure, ivermectin effects on these immune mechanisms were constant across ages, revealing no immunosenescence caused by ivermectin. Whereas males suffered a reduction in PO and proPO with ivermectin and age, female proPO was not affected by ivermectin or age. This reveals a strategy of self-care adopted by females to prioritize immune function when facing stressful conditions. Among sublethal effects caused by ivermectin, immunosuppression might be a main physiological driver of population declines in contaminated environments.

As critical pollinators of agricultural crops, honey bees (Apis mellifera) play a vital role in food production. Therefore, it is imperative to investigate and develop new methods to control Varroa destructor, a devastating parasitic mite of honey bee colonies that weakens bees and spreads deadly diseases that lead to colony loss. Here, we adapted existing methods to investigate the role of ABCB1 transporters in mitigating the toxicity of amitraz, a widely used miticide approved for use in honey bee colonies, demonstrating that a pharmacological inhibitor can synergistically increase amitraz toxicity compared to the equivalent dose of amitraz alone. Evaluations performed on the mites used in one of the described experiments revealed a high proportion of the test subject mites (88.5%) possessed the amitraz resistant genotype, indicating that even in resistant mites, inhibiting ABCB1 transporters can increase amitraz efficacy. This promising finding may be useful in developing powerful synergists that can be used to increase the efficacy of new and existent miticides.

Accurately determining the mating status of the agricultural fruit fly pest Ceratitis capitata, commonly known as Medfly, is essential for timely and effective eradication efforts. To overcome the limitations of subjective DAPI-based staining assessments of females captured in Jackson dry traps and Multilure liquid traps, we developed a multi-tier molecular diagnostic method that unequivocally detects mating status using DNA probes targeting the male-specific Y114 locus on the Y-chromosome of the species. Our protocol integrates morphological evaluation with increasingly sensitive molecular assays through the following steps: 1) A preliminary quality assessment of the specimens physical condition, DNA preservation, and mating status using conventional PCR followed by agarose electrophoresis (cPCR); 2) Quantification and real-time detection of sperm presence via quantitative PCR (qPCR); and 3) Detection of trace sperm amounts through droplet digital PCR (ddPCR). This PCR-based framework is designed for samples collected in the field, enabling accurate analysis of specimens exposed to adverse environmental conditions and varying levels of preservation after 2- and 3-weeks weathering times in traps. It allows quantitative determination of mating status even when sperm concentrations are extremely low, such as during transient copulation, and achieves detection limits down to approximately 14 spermatozoa in a mated female. By accounting for variable specimen quality and the performance characteristics of each molecular platform, this tiered approach ensures highly sensitive and unequivocal detection of mated females. The methodology can be used to assist eradication efforts across the C. capitata geographic range through the timely detection of mated females, halting their expansion and establishment into novel regions reducing control and eradication costs.

Huanglongbing (HLB, or citrus greening disease) affects all citrus varieties world-wide. In the USA, Asia, and South America the causal agent is "Candidatus Liberibacter asiaticus" (CLas), a phloem-limited, uncultured, alphaproteobacterium. The hemipteran insect vector, Diaphorina citri (Asian citrus psyllid) acquires and transmits CLas in a circulative, propagative manner. In addition to CLas, D. citri hosts multiple symbiotic bacterial species including Wolbachia (wDi). In D. citri, wDi has been sequenced and studied but specific roles in D. citri biology are unknown. Using well established quantitative PCR methods we measured CLas titer in D. citri collected from four groves in central Florida with distinct HLB management strategies and tested whether CLas and wDi titer were correlated in a sub-set of these insects. Grove site had the largest effect on CLas titer. Sex had no effect on CLas titer, while higher wDi titer was correlated with non-infected insects. Our results suggest that more directed follow-up research is necessary and important to clarify whether field management tactics influence CLas titer in D. citri and to better understand gene-by-environment interactions among D. citri, wDi and CLas. Now that millions of trees in Florida have been treated with injectable formulations of oxytetracycline, which is likely to decrease bacterial populations in D. citri, this study may represent the last biologically meaningful snapshot of grove-level vector-pathogen ecology in the state during the HLB epidemic.

Evidence-based decision making on malaria vector control strategies increasingly rely on triangulation of data which requires informatics systems that can integrate data from complex, multi-stage studies involving mosquitoes. This manuscript describes a performance evaluation of an extended version of the generic schema underpinning the VBDs360 platform, specifically improved to accommodate multiple distinct entomological assays spanning the field, insectary and laboratory. The utility of this extension, with respect to high-fidelity data linkage and robust sample traceability across complex entomological workflows, was evaluated through a case study conducted in southern Tanzania. Wild female mosquitoes were collected from 40 locations across a >4,000 km{superscript 2} area and then reared through multiple generations in an insectary before derived iso-female lineages were tested for phenotypic susceptibility to a pyrethroid insecticide. Such multi-generational lineages (F to F where n [&ge;] 2) were propagated to prevent non-heritable maternal effects on phenotype and produce enough progeny for standard WHO susceptibility assays. All samples were subsequently archived in a molecular laboratory, where all F specimens were tested for sibling species identity. A paper-based implementation of the extended schema enabled successful integration of 77,017 lines of data distributed across 6 different tables that spanned 3 distinct field, insectary, and laboratory workflows, implemented by three different teams working in different locations. At each step, fully independent and redundant primary and secondary keys enabled high fidelity error correction and sample tracing. Consistently perfect linkage between assay design and sample sorting data was achieved for F0 wild-caught adults, with 100% of 66,108 record successfully linked between field capture and morphological categorization. This complete traceability extended to the propagation of derived Fn lineages, with all 100 and 243 records from 9 adult-derived and 13 larval-derived lineages, respectively, correctly linked. Insecticide susceptibility phenotype further confirmed 100% linkage for 5,654 records between exposure history and recorded mortality outcome data in the insectary. Although such cross-cleaned linkages to sample analysis and storage data recorded by the laboratory team were not entirely perfect and could be improved, they were nevertheless of very high fidelity (97.3% (1967/2,022) for F0 samples and 99.3% (437/440) for Fn samples). Overall, this pilot application of the extended generic schema ensured robust data provenance and minimized transcription errors in this complex study distributed across multiple teams and locations. These findings demonstrate how this generic informatics framework may be scaled and adapted to support data integrity across diverse, large-scale, multi-team entomological research workflows.

Observing honeybee (Apis mellifera) brood dynamics and host-parasite interactions, especially within sealed cells, is vital for colony health research but traditionally relies on disruptive or destructive methods. Here we present BroodScan, a monitoring system employing a modified consumer-grade Charge-Coupled Device (CCD) flatbed scanner for continuous, high-resolution basal imaging of brood cells directly within an active beehive. Our experiments, conducted over a 35-day period, demonstrate BroodScans capacity to track the complete honeybee brood development cycle from egg to adult emergence, yielding quantitative data such as an average egg stage duration of 72.93 {+/-} 2.04 hours (n=58). Notably, the system provided detailed, time-lapse visual documentation of the complete intra-cellular reproductive cycle of the parasitic mite Varroa destructor within sealed worker cells, including the progression from foundress mite to multiple distinct offspring stages and their maturation. BroodScan offers a valuable, non-invasive tool that overcomes key limitations of previous techniques, opening new avenues for research into honeybee biology, colony health, and the impacts of diverse stressors like Varroa destructor, thereby promising to enhance our understanding and stewardship of these vital pollinators.