Management strategies are needed that mitigate the effects of climate change on honey bee colony losses. Extended periods of warmer fall temperatures prolong periods of honey bee flight and parasitic Varroa mite immigration into colonies. We report on a management strategy using Varroa-resistant Russian honey bees overwintered in indoor cold storage facilities, and compare colony survival and growth with that of unselected European bees. Fat body metrics that are key to overwintering survival were also measured in Russian and unselected bees. Comparisons between overwintering Russian colonies in cold storage versus apiaries were also conducted. Russian and unselected colonies overwintered in cold storage had comparable overwintering survival and percentages rented for almond pollination. However, more Russian colonies overwintered in cold storage were alive after almond bloom than those overwintered in apiaries. Fat bodies in Russian and unselected bees gained weight while in cold storage. Protein concentrations increased and lipids decreased. Changes in lipid concentrations were inversely related to the number of brood bees reared while in cold storage. Similar percentages of colonies overwintered in cold storage or outdoor apiaries survived and were rented for almond pollination. An economic analysis indicated that overwintering Russian colonies in cold storage costs less than in apiaries. Our study indicates that cold storage can be a viable management strategy for mitigating the effects of climate change on colony survival.

Vairimorpha (Microsporidia: Nosematidae) is a microsporidian that infects honey bees especially in winter. Fumagillin can reduce infections, but whether overwintering survival is improved is unclear. The diet also may influence the severity of Nosema infections. We examined the relationship between Nosema and colony size and survival in hives overwintered in cold storage facilities. In year 1, no Fumagillin treatments were applied. Colony size and survival after cold storage and almond bloom were comparable between groups with high and low pre-cold storage infections. In year 2, size and survival were compared among colonies with and without Fumagillin treatment that were fed either pollen or protein supplement prior to overwintering. Colonies treated with Fumagillin had lower spore numbers than untreated, but colony sizes and survival were similar among the treatments. However, more colonies with zero spores per bee could be rented for almond pollination and were alive after bloom than those averaging >1 million spores per bee. Fat body metrics can affect overwintering success. In both years, fat body weights and protein concentrations increased, and lipid concentrations decreased while bees were in cold storage. Fat body metrics did not differ with Nosema infection levels. However, Fumagillin negatively affected pre-cold storage fat body protein concentrations and colony sizes after cold storage and almond bloom. Treating with Fumagillin before overwintering in cold storage might result in greater colony survival if spore numbers are high, but undetectable or even negative effects when spore numbers are low.

For over a decade, high percentages of honey bee colonies have been perishing during the winter creating economic hardship to beekeepers and growers of early-season crops requiring pollination. A way to reduce colony losses might be moving hives into cold storage facilities for the winter. We explored factors that could affect the size and survival of colonies overwintered in cold storage and then used for almond pollination. The factors were when hives were put into cold storage and their location prior to overwintering. We found that colonies summered in North Dakota, USA and moved to cold storage in October were larger after cold storage and almond pollination than those moved in November. Colony location prior to overwintering also affected size and survival. Colonies summered in southern Texas, USA and moved to cold storage in November were smaller after cold storage and almond pollination than those from North Dakota. The colonies also were smaller than those overwintered in Texas apiaries. Fat body metrics of bees entering cold storage differed between summer locations. North Dakota bees had higher lipid and lower protein concentrations than Texas bees. While in cold storage, fat bodies gained weight, protein concentrations increased, and lipids decreased. The decrease in lipid concentrations was correlated with the amount of brood reared while colonies were in cold storage. Our study indicates that in northern latitudes, overwintering survival might be affected by when colonies are put into cold storage and that colonies summered in southern latitudes should be overwintered there.

Honey bee colonies have a yearly cycle that is supported nutritionally by the seasonal progression of flowering plants. In the spring, colonies grow by rearing brood, but in the fall, brood rearing declines in preparation for overwintering. Depending on where colonies are located, the yearly cycle can differ especially in overwintering activities. In temperate climates of Europe and North America, colonies reduce or end brood rearing in the fall while in warmer climates bees can rear brood and forage throughout the year. To test the hypothesis that nutrients available in seasonal pollens and honey bee responses to them can differ we analyzed pollen in the spring and fall collected by colonies in environments where brood rearing either stops in the fall (Iowa) or continues through the winter (Arizona). We fed both types of pollen to worker offspring of queens that emerged and open mated in each type of environment. We measured physiological responses to test if they differed depending on the location and season when the pollen was collected and the queen line of the workers that consumed it. Specifically, we measured pollen and protein consumption, gene expression levels (hex 70, hex 110, and vg) and hypopharyngeal gland (HPG) development. We found differences in macronutrient content and amino and fatty acids between spring and fall pollens from the same location and differences in nutrient content between locations during the same season. We also detected queen type and seasonal effects in HPG size and differences in gene expression between bees consuming spring vs. fall pollen with larger HPG and higher gene expression levels in those consuming spring pollen. The effects might have emerged from the seasonal differences in nutritional content of the pollens and genetic factors associated with the queen lines we used.

Varroa destructor is an ectoparasitic mite of immature and adult honey bees that can transmit several single-stranded RNA viruses to its host. Varroa reproduce in brood cells, and mite populations increase as colonies produce brood in spring and summer. Mite numbers also can sharply rise, particularly in the fall, by the migration of varroa into hives on foragers. Colonies with high levels of varroa and viruses often die over the winter. Feeding colonies pollen might keep virus levels low and improve survival because of the positive effects of pollen on immunity and colony growth. We compared varroa and virus levels and overwinter survival in colonies with (fed) and without (unfed) supplemental pollen. We also measured the frequency of capturing foragers with mites (FWM) at colony entrances to determine its relationship to varroa and virus levels. Colonies fed supplemental pollen were larger than unfed colonies and survived longer. Varroa populations and levels of Deformed wing virus (DWV) rose throughout the season, and were similar between fed and unfed colonies. The growth of varroa populations was correlated with FWM in fed and unfed colonies, and significantly affected DWV levels. Increasing frequencies of FWM and the effects on varroa populations might reduce the positive influence of supplemental pollen on immune function. However, pollen feeding can stimulate colony growth and this can improve colony survival.

Free-ranging herbivores have yearly life cycles that generate dynamic resource needs. Honey bee colonies also have a yearly life cycle that might generate nutritional requirements that differ between times of brood rearing and colony expansion in the spring and population contraction and preparation for overwintering in the fall. To test this, we analyzed polyfioral mixes of spring and fall pollens to determine if the nutrient composition differed with season. Next, we fed both types of seasonal pollens to bees reared in spring and fall. We compared the development of brood food glands (i.e., hypopharyngeal glands - HPG), and the expression of genes in the fat body between bees fed pollen from the same (in-season) or different season (out-of-season) when they were reared. Because pathogen challenges often heighten the effects of nutritional stress, we infected a subset of bees with Nosema to determine if bees responded differently to the infection depending on the seasonal pollen they consumed. We found that spring and fall pollens were similar in total protein and lipid concentrations, but spring pollens had higher concentrations of amino and fatty acids that support HPG growth and brood production. Bees responded differently when fed in vs. out of season pollen. The HPG of both uninfected and Nosema-infected spring bees were larger when they were fed spring (in-season) compared to fall pollen. Spring bees differentially regulated more than 200 genes when fed in- vs. out-of-season pollen. When infected with Nosema, approximately 400 genes showed different infection-induced expression patterns in spring bees depending on pollen type. In contrast, HPG size in fall bees was not affected by pollen type, though HPG were smaller in those infected with Nosema. Very few genes were differentially expressed with pollen type in uninfected (4 genes) and infected fall bees (5 genes). Pollen type did not affect patterns of infection-induced expression in fall bees. Our data suggest that physiological responses to seasonal pollens differ between bees reared in the spring and fall with spring bees being significantly more sensitive to pollen type especially when infected with Nosema. This study provides evidence that seasonal pollens may provide levels of nutrients that align with the activities of honey bees during their yearly colony cycle. The findings are important for the planning and establishment of forage plantings to sustain honey bees, and in the development of seasonal nutritional supplements fed to colonies when pollen is unavailable.

Honey bees that consume pollen with sublethal levels of the fungicide Pristine® can have reduced pollen digestion, lower ATP synthesis, and in many ways resemble malnourished bees. Reduced nutrient acquisition in bees exposed to Pristine® might be due to the effects of this fungicide on the composition of gut microbial communities. Two experiments were conducted in two separate years to test for the effects of Pristine® on the composition and diversity of bacteria in nurse bee midguts. In the low-dose experiment, bees fed the fungicide had a reduced relative abundance of Gilliamella sp. and the number of bacterial taxa within each individual bee was lower. In the high-dose experiment, the fungicide treatment led to increased relative abundance of Lactobacillus sp. Firm 4 and Firm 5 relative to the control. Presence of the fungicide did not impact the distribution of bacteria among individuals on either the low-or high-dose experiments. Considerable differences in gut microbial communities existed between the two experiments perhaps due to environmental and dietary factors. The effects of Pristine® on the gut microbiota were inconsistent between experiments indicating that exposure rate and environmental conditions can influence the effects of this fungicide on gut microbial communities.

Malnutrition is a major cause of colony losses. In managed hives, bees are fed protein supplements (PS) during pollen shortages. If bees were provided with natural forage instead of PS, would they have lower pathogen levels and higher queen and colony survival? We addressed this question by either providing colonies with forage (Brassica rapa-rapini) or feeding them PS from November to February. Soluble protein concentrations in the PS were lower than the rapini pollen as were levels of most amino acids. Nurse bees digested less of the protein in PS than the pollen. Hemolymph protein titers in nurse bees and colony growth did not differ between those fed PS or foraging on rapini. However, colonies fed PS had higher levels of black queen cell virus and Nosema and greater queen losses, indicating that natural forage might improve overwintering survival.

Varroa mites are a serious pest of honey bees and the leading cause of colony losses. Varroa have relatively low reproductive rates, so populations should not increase rapidly, but often they do. Other factors might contribute to the growth of varroa populations including mite migration into colonies on foragers from other hives. We measured the proportion of foragers carrying mites on their bodies while entering and leaving hives, and determined its relationship to the growth of varroa populations in those hives at two apiary sites. We also compared the estimates of mite population growth with predictions from a varroa population dynamics model that generates estimates of mite population growth based on mite reproduction. Samples of capped brood and adult bees indicated that the proportion of brood cells infested with mites and adult bees with phoretic mites was low through the summer but increased sharply in the fall especially at site 1. The frequency of capturing foragers with mites on their bodies while entering or leaving hives also increased in the fall. The growth of varroa populations at both sites was not significantly related to our colony estimates of successful mite reproduction, but instead to the total number of foragers with mites (entering and leaving the colony). There were more foragers with mites at site 1 than site 2, and mite populations at site 1 were larger especially in the fall. The model accurately estimated phoretic mite populations and infested brood cells until November when predictions were much lower than those measured in colonies. The rapid growth of mite populations particularly in the fall being a product of mite migration rather than mite reproduction only is discussed.

Sublethal exposure to fungicides can affect honey bees (Apis mellifera L.) in ways that resemble malnutrition. These include reduced brood rearing, queen loss, and increased pathogen levels. We examined the effects of oral exposure to the fungicides boscalid and pyraclostrobin on factors affecting colony nutrition and immune function including pollen consumption, protein digestion, hemolymph protein titers, and changes in virus levels. Because the fungicides are respiratory inhibitors, we also measured ATP concentrations in flight muscle. The effects were evaluated in 3- and 7-d-old worker bees at high fungicide concentrations in cage studies, and at field-relevant concentrations in colony studies. Though fungicide levels differed greatly between the cage and colony studies, similar effects were observed. Hemolymph protein concentrations were comparable between bees feeding on pollen with and without added fungicides. However, in both cage and colony studies, bees consumed less pollen containing fungicides and digested less of the protein. Bees fed fungicide-treated pollen also had lower ATP concentrations and higher virus titers. The combination of effects we detected could produce symptoms that are similar to those from poor nutrition and weaken colonies making them more vulnerable to loss from additional stressors such as parasites and pathogens.

Ten self-fertile commercial sunflowers cultivars were evaluated for seed set with and without exposure to bees. In the first planting, the number of foraging honey bees was smaller than in the second, and seed set for most cultivars did not differ between those bagged to exclude bees and ones that were open pollinated. In the second planting, however, a majority of cultivars had significantly greater seed set when capitula were exposed to bees compared with when they were not. The weight of seeds from open-pollinated capitula was greater than from those where bees were excluded. Environmental conditions also played a role in seed set as evidenced by differences between plantings in set on bagged capitula. In the first planting, average maximum and minimum temperatures were significantly higher than in the second, and overall seed set was significantly lower in capitula where bees were excluded compared with the second planting. Under the high temperature conditions, however, some cultivars set four times more seed on open-pollinated capitula compared with those that were bagged. These results suggest that foraging activity and cross-pollination by bees might mitigate reductions in seed set caused by high temperatures.