collection-and-identification-of-pollen-from-honey-bee-colonies

Collection and Identification of Pollen from Honey Bee Colonies

Researchers often collect and analyze corbicular pollen from honey bees to identify the plant sources on which they forage for pollen or to estimate ...

Oregon State University

In honey bees and many other social insects, production of queens is a vital task, as colony fitness is dependent on queens. The factors considered by honey bee workers in selecting larvae to rear new queens during emergency queen rearing are poorly understood. Identifying these parameters is critical, both in an evolutionary and apicultural context. As female caste development in honey bees is dependent on larval diet (i.e. nutrition), we hypothesized that larval nutritional state is meticulously assessed and used by workers in selection of larvae for queen rearing. To test this hypothesis, we conducted a series of experiments manipulating the nutritional status of one day old larvae by depriving them of brood food for a four-hour period, and then allowing workers to choose larvae for rearing queens from nutritionally deprived and non-deprived larvae. We simultaneously investigated the role of genetic relatedness in selection of larvae for queen rearing. In all the experiments, significantly greater numbers of non-deprived larvae than deprived larvae were selected for queen rearing irrespective of genetic relatedness. Our results demonstrate that honey bees perceive the nutritional state of larvae and use that information when selecting larvae for rearing queens in the natural emergency queen replacement process.

Honey bees consider larval nutritional status rather than genetic relatedness when selecting larvae for emergency queen rearing.

Global western honey bee, Apis mellifera (L.) (Hymenoptera: Apidae), colony declines pose a significant threat to food production worldwide. Poor nutrition resulting from habitat loss, extensive monocultures, and agricultural intensification is among the several suggested drivers for colony declines. Pollen is the primary source of protein for honey bees; therefore, both pollen abundance and diversity are critical for colony growth and survival. Many cropping systems that employ honey bee colonies for pollination may lack sufficient pollen diversity and abundance to provide optimal bee nutrition. In this observational study, we documented the diversity and relative abundance of pollen collected by honey bees in five major pollinator-dependent crops in the western United States. We sampled pollen from pollen traps installed on honey bee colonies in the following cropping systems-almond, cherry, highbush blueberry, hybrid carrot, and meadowfoam. The pollen diversity was estimated by documenting the number of different pollen pellet colors and plant taxa found in each pollen sample. The lowest pollen diversity was found in almond crop. Relatively higher quantities of pollen collection were collected in almond, cherry, and meadowfoam cropping systems. The information gleaned from this study regarding pollen diversity and abundance may help growers, land managers, and beekeepers improve pollen forage available to bees in these cropping systems.

Assessment of Pollen Diversity Available to Honey Bees (Hymenoptera: Apidae) in Major Cropping Systems During Pollination in the Western United States.

Significant annual honey bee colony losses have been reported in the USA and across the world over the past years. Malnutrition is one among several causative factors for such declines. Optimal nutrition serves as the first line of defense against multiple stressors such as parasites/pathogens and pesticides. Given the importance of nutrition, it is imperative to understand bee nutrition holistically, identifying dietary sources that may fulfill bee nutritional needs. Pollen is the primary source of protein for bees and is critical for brood rearing and colony growth. Currently, there is significant gap in knowledge regarding the chemical and nutritional composition of pollen.

The omics approach to bee nutritional landscape.

The decline in populations of insect pollinators is a global concern. While multiple factors are implicated, there is uncertainty surrounding the contribution of certain groups of pesticides to losses in wild and managed bees. Nanotechnology-based pesticides (NBPs) are formulations based on multiple particle sizes and types. By packaging active ingredients in engineered particles, NBPs offer many benefits and novel functions, but may also exhibit different properties in the environment when compared with older pesticide formulations. These new properties raise questions about the environmental disposition and fate of NBPs and their exposure to pollinators. Pollinators such as honey bees have evolved structural adaptations to collect pollen, but also inadvertently gather other types of environmental particles which may accumulate in hive materials. Knowledge of the interaction between pollinators, NBPs, and other types of particles is needed to better understand their exposure to pesticides, and essential for characterizing risk from diverse environmental contaminants. The present review discusses the properties, benefits and types of nanotechnology-based pesticides, the propensity of bees to collect such particles and potential impacts on bee pollinators.

Potential Risk to Pollinators from Nanotechnology-Based Pesticides.

Poor nutrition is an important factor in global bee population declines. A significant gap in knowledge persists regarding the role of various nutrients (especially micronutrients) in honey bees. Sterols are essential micronutrients in insect diets and play a physiologically vital role as precursors of important molting hormones and building blocks of cellular membranes. Sterol requirements and metabolism in honey bees are poorly understood. Among all pollen sterols, 24-methylenecholesterol is considered the key phytosterol required by honey bees. Nurse bees assimilate this sterol from dietary sources and store it in their tissues as endogenous sterol, to be transferred to the growing larvae through brood food. This study examined the duration of replacement of such endogenous sterols in honey bees. The dietary C-labeled isotopomer of 24-methylenecholesterol added to artificial bee diet showed differential, progressive in vivo assimilation across various honey bee tissues. Significantly higher survival, diet consumption, head protein content and abdominal lipid content were observed in the dietary sterol-supplemented group than in the control group. These findings provide novel insights into phytosterol utilization and temporal pattern of endogenous 24-methylenecholesterol replacement in honey bees.

Novel Insights into Dietary Phytosterol Utilization and Its Fate in Honey Bees ( L.).

Although poor nutrition is cited as one of the crucial factors in global pollinator decline, the requirements and role of several important nutrients (especially micronutrients) in honey bees are not well understood. Micronutrients, viz. phytosterols, play a physiologically vital role in insects as precursors of important molting hormones and building blocks of cellular membranes. There is a gap in comprehensive understanding of the impacts of dietary sterols on honey bee physiology. In the present study, we investigated the role of 24-methylenecholesterol-a key phytosterol-in honey bee nutritional physiology. Artificial diets with varying concentrations of 24-methylenecholesterol (0%, 0.1%. 0.25%, 0.5%, 0.75%, and 1% dry diet weight) were formulated and fed to honey bees in a laboratory cage experiment. Survival, diet consumption, head protein content, and abdominal lipid contents were significantly higher in dietary sterol-supplemented bees. Our findings provide additional insights regarding the role of this important sterol in honey bee nutritional physiology. The insights gleaned from this study could also advance the understanding of sterol metabolism and regulation in other bee species that are dependent on pollen for sterols, and assist in formulation of a more complete artificial diet for honey bees ( Linnaeus, 1758) (Hymenoptera: Apidae).

Evaluating Effects of a Critical Micronutrient (24-Methylenecholesterol) on Honey Bee Physiology.

Pesticide exposures can have detrimental impacts on bee pollinators, ranging from immediate mortality to sub-lethal impacts. Flupyradifurone is the active ingredient in Sivanto™ and sulfoxaflor is the active ingredient in Transform®. They are both relatively new insecticides developed with an intent to reduce negative effects on bees, when applied to bee-attractive crops. With the growing concern regarding pollinator health and pollinator declines, it is important to have a better understanding of any potential negative impacts, especially sub-lethal, of these pesticides on bees. This study reports novel findings regarding physiological stress experienced by bees exposed to field application rates of these two insecticides via a Potter Tower sprayer. Two contact exposure experiments were conducted-a shorter 6-hour study and a longer 10-day study. Honey bee mortality, sugar syrup and water consumption, and physiological responses (oxidative stress and apoptotic protein assays) were assessed in bees exposed to Sivanto™ and Transform®, and compared to bees in control group. For the longer, 10-day contact exposure experiment, only the Sivanto™ group was compared to the control group, as high mortality recorded in the sulfoxaflor treatment group during the shorter contact exposure experiment, made the latter group unfeasible to test in the longer 10-days experiment. In both the studies, sugar syrup and water consumptions were significantly different between treatment groups and controls. The highest mortality was observed in Transform® exposed bees, followed by the Sivanto™ exposed bees. Estimates of reactive oxygen/nitrogen species indicated significantly elevated oxidative stress in both pesticide treatment groups, when compared to controls. Caspase-3 protein assays, an indicator of onset of apoptosis, was also significantly higher in the pesticide treatment groups. These differences were largely driven by post exposure duration, indicating sub-lethal impacts. Further, our findings also emphasize the need to revisit contact exposure impacts of Sivanto™, given the sub-lethal impacts and mortality observed in our long-term (10-day) contact exposure experiment.

Field rates of Sivanto™ (flupyradifurone) and Transform® (sulfoxaflor) increase oxidative stress and induce apoptosis in honey bees (Apis mellifera L.).

Phytosterols are important micronutrients that are precursors of important molting hormones and help maintain cellular membrane integrity in insects including bees. Previous research has shown that 24-methylenecholesterol is a key phytosterol that enhances honey bee longevity and improves nurse bee physiology. Nurse bees have the ability to selectively transfer this sterol to developing larvae through brood food. This study examines the physiological impacts of 24-methylenecholesterol on nurse bees, by analyzing the protein profiles of nurse bee heads upon dietary sterol manipulation. Dietary experimental groups consisting of newly emerged honey bees were provided with varying concentrations of 24-methylenecholesterol for three weeks. At the end of the study, honey bees were collected and proteomic analysis was performed on honey bee heads. A total of 1715 proteins were identified across experimental groups. The mean relative abundances of nutritional marker proteins (. major royal jelly proteins 1, 4, 5, 7) were higher in experimental groups supplemented with higher dietary sterol concentrations, when compared with the control dietary group. The mean relative abundances of important enzymatic proteins (aminopeptidase and calcium-transporting ATPase) were higher in control groups, whereas mean relative abundances of oxysterol-binding protein and fatty acid-binding protein were higher in higher dietary sterol groups.

Changes in Honey Bee Head Proteome in Response to Dietary 24-Methylenecholesterol.

Honey bees provision glandular secretions in the form of royal jelly as larval nourishment to developing queens. Exposure to chemicals and nutritional conditions can influence queen development and thus impact colony fitness. Previous research reports that royal jelly remains pesticide-free during colony-level exposure and that chemical residues are buffered by the nurse bees. However, the impacts of pesticides can also manifest in quality and quantity of royal jelly produced by nurse bees. Here, we tested how colony exposure to a multi-pesticide pollen treatment influences the amount of royal jelly provisioned per queen and the additional impacts on royal jelly nutritional quality. We observed differences in the metabolome, proteome, and phytosterol compositions of royal jelly synthesized by nurse bees from multi-pesticide exposed colonies, including significant reductions of key nutrients such as 24-methylenecholesterol, major royal jelly proteins, and 10-hydroxy-2-decenoic acid. Additionally, quantity of royal jelly provisioned per queen was lower in colonies exposed to pesticides, but this effect was colony-dependent. Pesticide treatment had a greater impact on royal jelly nutritional composition than the weight of royal jelly provisioned per queen cell. These novel findings highlight the indirect effects of pesticide exposure on queen developmental nutrition and allude to social consequences of nurse bee glandular degeneration.

Colony-level pesticide exposure affects honey bee (Apis mellifera L.) royal jelly production and nutritional composition.

Global decline in insect pollinators, especially bees, have resulted in extensive research into understanding the various causative factors and formulating mitigative strategies. For commercial beekeepers in the United States, overwintering honey bee colony losses are significant, requiring tactics to overwinter bees in conditions designed to minimize such losses. This is especially important as overwintered honey bees are responsible for colony expansion each spring, and overwintered bees must survive in sufficient numbers to nurse the spring brood and forage until the new 'replacement' workers become fully functional. In this study, we examined the physiology of overwintered (diutinus) bees following various overwintering storage conditions. Important physiological markers, i.e., head proteins and abdominal lipid contents were higher in honey bees that overwintered in controlled indoor storage facilities, compared with bees held outdoors through the winter months. Our findings provide new insights into the physiology of honey bees overwintered in indoor and outdoor environments and have implications for improved beekeeping management.

Article	Year
Honey bees consider larval nutritional status rather than genetic relatedness when selecting larvae for emergency queen rearing. Scientific reports\| PubMed ID: 29769574	2018
Assessment of Pollen Diversity Available to Honey Bees (Hymenoptera: Apidae) in Major Cropping Systems During Pollination in the Western United States. Journal of economic entomology\| PubMed ID: 31237612	2019
The omics approach to bee nutritional landscape. Metabolomics : Official journal of the Metabolomic Society\| PubMed ID: 31538263	2019
Potential Risk to Pollinators from Nanotechnology-Based Pesticides. Molecules (Basel, Switzerland)\| PubMed ID: 31817417	2019
Novel Insights into Dietary Phytosterol Utilization and Its Fate in Honey Bees ( L.). Molecules (Basel, Switzerland)\| PubMed ID: 32012964	2020
Evaluating Effects of a Critical Micronutrient (24-Methylenecholesterol) on Honey Bee Physiology. Annals of the Entomological Society of America\| PubMed ID: 32410742	2020
Field rates of Sivanto™ (flupyradifurone) and Transform® (sulfoxaflor) increase oxidative stress and induce apoptosis in honey bees (Apis mellifera L.). PloS one\| PubMed ID: 32437365	2020
Changes in Honey Bee Head Proteome in Response to Dietary 24-Methylenecholesterol. Insects\| PubMed ID: 33138161	2020
Colony-level pesticide exposure affects honey bee (Apis mellifera L.) royal jelly production and nutritional composition. Chemosphere\| PubMed ID: 33297150	2021
Impacts of Different Winter Storage Conditions on the Physiology of Diutinus Honey Bees (Hymenoptera: Apidae). Journal of economic entomology\| PubMed ID: 33386734	2021

Priyadarshini Chakrabarti

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