Honey bee colony behavior and ontogeny are adversely affected when exposed to a pesticide-contaminated environment.

Honey bees exhibit age polyethism and thus have a predictable sequence of behaviors they express through developmental time. Numerous laboratory studies show exposure to pesticides may impair critical honey bee behaviors (brood care, foraging, egg-laying, etc.) that adversely affect colony productivity and survival. There are fewer studies that examine the impacts of pesticides in natural field settings, especially given the challenges of implementing treatment groups and controlling variables. This study helps address the need for impact studies on pollinators under field conditions to assess the consequences of chemical overuse and dependency in agricultural and urban landscapes. To assess the impact of systemic pesticides in a natural field setting on worker bee behavioral development, observation hives were established to monitor changes in behaviors of similarly aged workers and sister queens within 2 experimental groups: (i) colonies located near point-source systemic pesticide pollution (pesticide contaminated treatment), and (ii) colonies embedded within a typical Midwestern US agricultural environment (control). In this study, worker bees in the contaminated environment exhibited important and biologically significant behavioral differences and accelerated onset of hive tasks (i.e., precocious behavioral development) compared to similarly aged bees at the control site. Queen locomotion was largely unaffected; however, the egg-laying rate was reduced in queens at the contaminated (treated) site. These results show that environmental pesticide exposure can disrupt colony function and adversely affect worker bee behavioral maturation, leading to reduced worker longevity and decreased colony efficiency.

Re-using food resources from failed honey bee (Apis mellifera L.) colonies and their impact on colony queen rearing capacity.

For over a decade, beekeepers have experienced high losses of honey bee (Apis mellifera L.) colonies due to a variety of stressors including pesticide exposure. Some of these chemical stressors may residually remain in the colony comb and food resources (pollen and nectar) of failed colonies and be later re-used by beekeepers when splitting and building back new colonies. The practice of re-using comb from previously perished colonies (termed "deadout") is common in beekeeping practice, but its role in affecting colony health is not well understood. Here, we evaluate the impact of reused, pesticide-contaminated "deadout" combs on colony function during the process of replacing a queen bee. Queenless microcolonies were established to monitor queen rearing capacity in two treatment groups: (1) colonies given frames containing food resources from deadout colonies in control "clean" apiaries and, (2) colonies given frames containing "contaminated" resources from deadout colonies originating from apiaries experiencing chronic pesticide exposure from widespread systemic pesticide pollution (including neonicotinoid insecticides: clothianidin and thiamethoxam). Results indicate that colonies given pesticide-contaminated resources produced fewer queen cells per colony and had a lower proportion of colonies successfully raising a functional, diploid egg-laying queen. This research highlights the deleterious effects of re-using deadout combs from colonies previously lost due to pesticide contamination.

Targeted Method for Quantifying Air-Borne Pesticide Residues from Conventional Seed Coat Treatments to Better Assess Exposure Risk During Maize Planting.

Agricultural seed-coat treatments are prone to drift as seed coatings may scuff off and become incorporated into field particles during planting. Vacuum planters release exhaust and kick up field dust, laden with systemic pesticides that blow across the landscape, is taken up, and later expressed in the nectar and pollen of surrounding plants. Offsite movements and nontarget exposure to systemic pesticides need attention and determining how and at what exposure levels pollinators are exposed is of critical importance. Unfortunately, this requires extensive and costly instrumental analyses. Here, we describe dust sampling and a modified, rapid method based on liquid chromatography in tandem with mass spectrometry-based method for quantification of a broad array of agrochemicals in captured dust particles. This method increases ability to detect potential exposure to multiple agrochemicals and allows researchers to better address critical knowledge gaps in the environmental fate, off-target movement, and persistence of conventional seed treatments.

Identification of anti-inflammatory vesicle-like nanoparticles in honey.

Honey has been used as a nutrient, an ointment, and a medicine worldwide for many centuries. Modern research has demonstrated that honey has many medicinal properties, reflected in its anti-microbial, anti-oxidant, and anti-inflammatory bioactivities. Honey is composed of sugars, water and a myriad of minor components, including minerals, vitamins, proteins and polyphenols. Here, we report a new bioactive component‒vesicle-like nanoparticles‒in honey (H-VLNs). These H-VLNs are membrane-bound nano-scale particles that contain lipids, proteins and small-sized RNAs. The presence of plant-originated plasma transmembrane proteins and plasma membrane-associated proteins suggests the potential vesicle-like nature of these particles. H-VLNs impede the formation and activation of the nucleotide-binding domain and leucine-rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome, which is a crucial inflammatory signalling platform in the innate immune system. Intraperitoneal administration of H-VLNs in mice alleviates inflammation and liver damage in the experimentally induced acute liver injury. miR-4057 in H-VLNs was identified in inhibiting NLRP3 inflammasome activation. Together, our studies have identified anti-inflammatory VLNs as a new bioactive agent in honey.

An updated genetic marker for detection of Lake Sinai Virus and metagenetic applications.

BACKGROUND: Lake Sinai Viruses (LSV) are common RNA viruses of honey bees (Apis mellifera) that frequently reach high abundance but are not linked to overt disease. LSVs are genetically heterogeneous and collectively widespread, but despite frequent detection in surveys, the ecological and geographic factors structuring their distribution in A. mellifera are not understood. Even less is known about their distribution in other species. Better understanding of LSV prevalence and ecology have been hampered by high sequence diversity within the LSV clade. METHODS: Here we report a new polymerase chain reaction (PCR) assay that is compatible with currently known lineages with minimal primer degeneracy, producing an expected 365 bp amplicon suitable for end-point PCR and metagenetic sequencing. Using the Illumina MiSeq platform, we performed pilot metagenetic assessments of three sample sets, each representing a distinct variable that might structure LSV diversity (geography, tissue, and species). RESULTS: The first sample set in our pilot assessment compared cDNA pools from managed A. mellifera hives in California (n = 8) and Maryland (n = 6) that had previously been evaluated for LSV2, confirming that the primers co-amplify divergent lineages in real-world samples. The second sample set included cDNA pools derived from different tissues (thorax vs. abdomen, n = 24 paired samples), collected from managed A. mellifera hives in North Dakota. End-point detection of LSV frequently differed between the two tissue types; LSV metagenetic composition was similar in one pair of sequenced samples but divergent in a second pair. Overall, LSV1 and intermediate lineages were common in these samples whereas variants clustering with LSV2 were rare. The third sample set included cDNA from individual pollinator specimens collected from diverse landscapes in the vicinity of Lincoln, Nebraska. We detected LSV in the bee Halictus ligatus (four of 63 specimens tested, 6.3%) at a similar rate as A. mellifera (nine of 115 specimens, 7.8%), but only one H. ligatus sequencing library yielded sufficient data for compositional analysis. Sequenced samples often contained multiple divergent LSV lineages, including individual specimens. While these studies were exploratory rather than statistically powerful tests of hypotheses, they illustrate the utility of high-throughput sequencing for understanding LSV transmission within and among species.

Proceedings of the 2020 American Bee Research Conference.

The 2020 American Bee Research Conference (ABRC) was held on 9-10 January 2020 in conjunction with the annual convention of the American Beekeeping Federation Conference and Trade Show in Schaumburg, IL. Over the two-day conference, a total of 65 oral and poster presentations were given, representing work done from over 30 different research groups from throughout the United States and Canada. These proceedings contain the submitted abstracts for presentations given at the 2020 American Bee Research Conference.

Comparative analysis of viruses in four bee species collected from agricultural, urban, and natural landscapes.

Managed honey bees (Apis mellifera L.) and wild bees provide critical ecological services that shape and sustain natural, agricultural, and urban landscapes. In recent years, declines in bee populations have highlighted the importance of the pollination services they provide and the need for more research into the reasons for global bee losses. Several stressors cause declining populations of managed and wild bee species such as habitat degradation, pesticide exposure, and pathogens. Viruses, which have been implicated as a key stressor, are able to infect a wide range of species and can be transmitted both intra- and inter-specifically from infected bee species to uninfected bee species via vertical (from parent to offspring) and/or horizontal (between individuals via direct or indirect contact) transmission. To explore how viruses spread both intra- and inter-specifically within a community, we examined the impact of management, landscape type, and bee species on the transmission of four common viruses in Nebraska: Deformed wing virus (DWV), Israeli acute paralysis virus (IAPV), Black queen cell virus (BQCV), and Sacbrood virus (SBV). Results indicated the prevalence of viruses is significantly affected (P < 0.005) by bee species, virus type, and season, but not by landscape or year (P = 0.290 and 0.065 respectively). The higher prevalence of DWV detected across bee species (10.4% on Apis mellifera, 5.3% on Bombus impatiens, 6.1% on Bombus griseocollis, and 22.44% on Halictus ligatus) and seasons (10.8% in early-mid summer and 11.4% in late summer) may indicate a higher risk of interspecific transmission of DWV. However, IAPV was predominately detected in Halictus ligatus (20.7%) and in late season collections (28.1%), which may suggest species-specific susceptibility and seasonal trends in infection rates associated with different virus types. However, there were limited detections of SBV and BQCV in bees collected during both sampling periods, indicating SBV and BQCV may be less prevalent among bee communities in this area.

Proceedings of the 2019 American Bee Research Conference.

The 2019 American Bee Research Conference (ABRC) was held January 10-12, 2019 in conjunction with the annual convention of the American Honey Producers Association in Tempe, AZ. Over the three-day conference, a total of 45 oral presentations and 13 poster presentations were given, representing work done from over 27 institutions and 34 different research groups from throughout the United States and Canada. This proceedings contains and overview of the conference and the submitted abstracts for presentations given at the 2018 American Bee Research Conference.

Interactions between pesticides and pathogen susceptibility in honey bees.

There exist a variety of factors that negatively impact the health and survival of managed honey bee colonies, including the spread of parasites and pathogens, loss of habitat, reduced availability or quality of food resources, climate change, poor queen quality, changing cultural and commercial beekeeping practices, as well as exposure to agricultural and apicultural pesticides both in the field and in the hive. These factors are often closely intertwined, and it is unlikely that a single stressor is driving colony losses. There is a growing consensus, however, that increasing prevalence of parasites and pathogens are among the most significant threats to managed bee colonies. Unfortunately, improper management of hives by beekeepers may exacerbate parasite populations and disease transmission. Furthermore, research continues to accumulate that describes the complex and largely harmful interactions that exist between pesticide exposure and bee immunity. This brief review summarizes our progress in understanding the impact of pesticide exposure on bees at the individual, colony, and community level.

Effects of neonicotinoid imidacloprid exposure on bumble bee (Hymenoptera: Apidae) queen survival and nest initiation.

Neonicotinoids are highly toxic to insects and may systemically translocate to nectar and pollen of plants where foraging bees may become exposed. Exposure to neonicotinoids can induce detrimental sublethal effects on individual and colonies of bees and may have long-term impacts, such as impaired foraging, reduced longevity, and reduced brood care or production. Less well-studied are the potential effects on queen bumble bees that may become exposed while foraging in the spring during colony initiation. This study assessed queen survival and nest founding in caged bumble bees [Bombus impatiens (Cresson) (Hymenoptera: Apidae)] after chronic (18-d) dietary exposure of imidacloprid in syrup (1, 5, 10, and 25 ppb) and pollen (0.3, 1.7, 3.3, and 8.3 ppb), paired respectively. Here we show some mortality in queens exposed at all doses even as low as 1 ppb, and, compared with untreated queens, significantly reduced survival of treated queens at the two highest doses. Queens that survived initial imidacloprid exposure commenced nest initiation; however, they exhibited dose-dependent delay in egg-laying and emergence of worker brood. Furthermore, imidacloprid treatment affected other parameters such as nest and queen weight. This study is the first to show direct impacts of imidacloprid at field-relevant levels on individual B. impatiens queen survival and nest founding, indicating that bumble bee queens are particularly sensitive to neonicotinoids when directly exposed. This study also helps focus pesticide risk mitigation efforts and highlights the importance of reducing exposure rates in the early spring when bumble bee queens, and other wild bees are foraging and initiating nests.

Weight of evidence evaluation of a network of adverse outcome pathways linking activation of the nicotinic acetylcholine receptor in honey bees to colony death.

Ongoing honey bee (Apis mellifera) colony losses are of significant international concern because of the essential role these insects play in pollinating crops. Both chemical and non-chemical stressors have been implicated as possible contributors to colony failure; however, the potential role(s) of commonly-used neonicotinoid insecticides has emerged as particularly concerning. Neonicotinoids act on the nicotinic acetylcholine receptors (nAChRs) in the central nervous system to eliminate pest insects. However, mounting evidence indicates that neonicotinoids also may adversely affect beneficial pollinators, such as the honey bee, via impairments on learning and memory, and ultimately foraging success. The specific mechanisms linking activation of the nAChR to adverse effects on learning and memory are uncertain. Additionally, clear connections between observed impacts on individual bees and colony level effects are lacking. The objective of this review was to develop adverse outcome pathways (AOPs) as a means to evaluate the biological plausibility and empirical evidence supporting (or refuting) the linkage between activation of the physiological target site, the nAChR, and colony level consequences. Potential for exposure was not a consideration in AOP development and therefore this effort should not be considered a risk assessment. Nonetheless, development of the AOPs described herein has led to the identification of research gaps which, for example, may be of high priority in understanding how perturbation of pathways involved in neurotransmission can adversely affect normal colony functions, causing colony instability and subsequent bee population failure. A putative AOP network was developed, laying the foundation for further insights as to the role of combined chemical and non-chemical stressors in impacting bee populations. Insights gained from the AOP network assembly, which more realistically represents multi-stressor impacts on honey bee colonies, are promising toward understanding common sensitive nodes in key biological pathways and identifying where mitigation strategies may be focused to reduce colony losses.

Sub-lethal effects of dietary neonicotinoid insecticide exposure on honey bee queen fecundity and colony development.

Many factors can negatively affect honey bee (Apis mellifera L.) health including the pervasive use of systemic neonicotinoid insecticides. Through direct consumption of contaminated nectar and pollen from treated plants, neonicotinoids can affect foraging, learning, and memory in worker bees. Less well studied are the potential effects of neonicotinoids on queen bees, which may be exposed indirectly through trophallaxis, or food-sharing. To assess effects on queen productivity, small colonies of different sizes (1500, 3000, and 7000 bees) were fed imidacloprid (0, 10, 20, 50, and 100 ppb) in syrup for three weeks. We found adverse effects of imidacloprid on queens (egg-laying and locomotor activity), worker bees (foraging and hygienic activities), and colony development (brood production and pollen stores) in all treated colonies. Some effects were less evident as colony size increased, suggesting that larger colony populations may act as a buffer to pesticide exposure. This study is the first to show adverse effects of imidacloprid on queen bee fecundity and behavior and improves our understanding of how neonicotinoids may impair short-term colony functioning. These data indicate that risk-mitigation efforts should focus on reducing neonicotinoid exposure in the early spring when colonies are smallest and queens are most vulnerable to exposure.