Cold storage as part of a Varroa management strategy: effects on honey bee colony performance, mite levels and stress biomarkers.

Placing honey bee colonies in cold storage has been proposed as a way to induce a pause in brood production as part of a Varroa mite treatment plan. Here, we exposed colonies to combinations of with or without an October cold storage period and with or without a subsequent miticide application. We then measured the effects of those treatments on colony-level variables (i.e. colony size, Varroa infestation level, survivorship and hive weight and temperature) and pooled individual-level variables that are associated with nutritional and stress responses. Colonies were assessed before and after cold storage, and again post winter, for a total duration of about 5 months, and the experiment was repeated. Brood levels were significantly lower after cold storage, and hive temperatures indicated that most or all brood had emerged after about two weeks in cold storage. However, Varroa levels at the end of the experiments in February were not significantly different among treatment groups. Colonies kept outside (not subjected to cold storage) and treated with a miticide had higher survivorship on average than any other treatment group, but no other group comparisons were significant, and long-term impact of cold storage on adult bee populations and on colony thermoregulation was low. The bee forage environment was also very different between the 2 years of the study, as rainfall and bee forage availability were much higher the second year. Colonies were over 2.5 times larger on average the second year compared to the first, both in terms of adult bee mass and brood area, and expression levels of nutrition and stress response genes were also significantly higher the second year. The results indicate that limited cold storage would likely have little long-term impact on most colony and individual measures of health, but for such a strategy to succeed levels of stressors, such as Varroa, may also need to be low.

Field and Cage Studies Show No Effects of Exposure to Flonicamid on Honey Bees at Field-Relevant Concentrations.

The extent to which insecticides harm non-target beneficial insects is controversial. The effects of long-term exposure on honey bees to sublethal concentrations of flonicamid, a pyridinecarboxamide compound used as a systemic insecticide against sucking insects, were examined in a field study and two cage studies. The field study involved the continuous weight, temperature, and CO2 monitoring of 18 honey bee colonies, 6 of which were exposed over six weeks to 50 ppb flonicamid in sugar syrup, 6 exposed to 250 ppb flonicamid in syrup, and 6 exposed to unadulterated syrup (control). Treatments were derived from concentrations observed in honey samples in a published study. No effects were observed on foraging activity, hive weight gain, thermoregulation, or average CO2 concentrations. However, Varroa mite infestations may have also contributed to experimental variability. The two cage studies, in which cages (200 newly-emerged bees in each) were exposed to the same flonicamid concentrations as the field study and kept in a variable-temperature incubator, likewise did not show any experiment-wide effects on survivorship, thermoregulation, or syrup consumption. These results suggest that field applications of flonicamid that result in concentrations as high as 250 ppb in honey may be largely safe for honey bees.

Integration of Scales and Cameras in Nondisruptive Electronic Beehive Monitoring: On the Within-Day Relationship of Hive Weight and Traffic in Honeybee (Apis mellifera) Colonies in Langstroth Hives in Tucson, Arizona, USA.

The relationship between beehive weight and traffic is a fundamental open research problem for electronic beehive monitoring and digital apiculture, because weight and traffic affect many aspects of honeybee (Apis mellifera) colony dynamics. An investigation of this relationship was conducted with a nondisruptive two-sensor (scale and camera) system on the weight and video data collected on six Apis mellifera colonies in Langstroth hives at the USDA-ARS Carl Hayden Bee Research Center in Tucson, Arizona, USA, from 15 May to 15 August 2021. Three hives had positive and two hives had negative correlations between weight and traffic. In one hive, weight and traffic were uncorrelated. The strength of the correlation between weight and traffic was stronger for longer time intervals. The traffic spread and mean, when taken separately, did not affect the correlation between weight and traffic more significantly than the exact traffic counts from videos. Lateral traffic did not have a significant impact on weight.

Traces of a neonicotinoid pesticide stimulate different honey bee colony activities, but do not increase colony size or longevity.

How neonicotinoid contamination affects honey bees remains controversial. Studies have yielded contradictory results, and few have examined effects on colony development. Here we report the results of a comprehensive five-year study of the effects of the neonicotinoid imidacloprid on honey bee colonies. Colonies fed 5 ng/g (ppb) imidacloprid in sugar syrup showed increased brood production, lower temperature variability, higher CO2 production and had more foragers compared to control colonies fed unadulterated syrup, but treatment did not affect adult bee numbers or average hive temperatures, and did not increase food stores, daily food acquisition or colony survivorship. These results suggest that imidacloprid contamination increased colony metabolism without improving colony productivity, and helps explain why some studies have reported no, or even positive, effects of neonicotinoids. Effect sizes were generally small but that could be attributed at least in part to variability in uncontrolled factors such as weather. We provide an explanation for the diverse effects of pesticide contamination on honey bees, and an improved understanding of how colonies are impacted.

Sublethal concentrations of clothianidin affect honey bee colony growth and hive CO2 concentration.

The effects of agricultural pesticide exposure upon honey bee colonies is of increasing interest to beekeepers and researchers, and the impact of neonicotinoid pesticides in particular has come under intense scrutiny. To explore potential colony-level effects of a neonicotinoid pesticide at field-relevant concentrations, honey bee colonies were fed 5- and 20-ppb concentrations of clothianidin in sugar syrup while control colonies were fed unadulterated syrup. Two experiments were conducted in successive years at the same site in southern Arizona, and one in the high rainfall environment of Mississippi. Across all three experiments, adult bee masses were about 21% lower among colonies fed 20-ppb clothianidin than the untreated control group, but no effects of treatment on brood production were observed. Average daily hive weight losses per day in the 5-ppb clothianidin colonies were about 39% lower post-treatment than in the 20-ppb clothianidin colonies, indicating lower consumption and/or better foraging, but the dry weights of newly-emerged adult bees were on average 6-7% lower in the 5-ppb group compared to the other groups, suggesting a nutritional problem in the 5-ppb group. Internal hive CO2 concentration was higher on average in colonies fed 20-ppb clothianidin, which could have resulted from greater CO2 production and/or reduced ventilating activity. Hive temperature average and daily variability were not affected by clothianidin exposure but did differ significantly among trials. Clothianidin was found to be, like imidacloprid, highly stable in honey in the hive environment over several months.

Landscape factors influencing honey bee colony behavior in Southern California commercial apiaries.

Colony brood levels, frames of bees (adult bee mass) and internal hive temperature were monitored for 60 colonies for each of two years as they were moved from agricultural, tree crop and mountain landscapes in southern California to blueberry and almond pollination sites. Hive weight was also continuously monitored for 20 of those hives for 6 weeks for both years, during commercial pollination. Pesticide residues in wax, honey and beebread samples were analyzed by composite apiary samples. While colonies in mountain sites had more adult bees and brood than those in agricultural sites in August, by October brood levels were higher in colonies from agricultural sites. Though hives from different original landscapes differed in size in October, hive assessments revealed no differences between the groups after co-wintering when graded for commercial almond pollination. Beebread from hives in agricultural sites had greater agrochemical diversity and in general higher pesticide hazard quotients than those from mountain sites, but those hives also had higher and more constant temperatures from September until January than hives from mountain sites. Hives placed in commercial almond pollination gained on average 287 g per d, compared to an average loss of 68 g per d for colonies in commercial blueberry pollination, although weight data indicated greater foraging effort by colonies in blueberries, possibly due to the proximity and abundance of almond pollen during bloom. Temperature monitoring was effective at distinguishing hive groups and had the best overall value in terms of equipment, installation, colony disturbance and information yield.

Exposure to sublethal concentrations of methoxyfenozide disrupts honey bee colony activity and thermoregulation.

Methoxyfenozide is an insect growth regulator (IGR) commonly used in agriculture to simultaneously control pests and preserve beneficial insect populations; however, its impact on honey bees in not fully understood. We conducted field and laboratory experiments to investigate bee health in response to field-relevant concentrations of this pesticide. Significant effects were observed in honey bee colony flight activity and thermoregulation after being exposed over 9 weeks to supplemental protein patty containing methoxyfenozide. Compared to bee colonies in the control group, colonies fed pollen patty with 200 ppb methoxyfenozide (as measured by residue analysis) had: 1) a significantly reduced rate of weight loss due to forager departure in the morning; and 2) higher temperature variability during the winter. Colonies in the 100 ppb (as measured by residue analysis) treatment group had values between the 200 ppb group and control for both response variables. The dusk break point, which is the time associated with the end of forager return, differed among all treatment groups but may have been confounded with direction the hives were facing. Bee colony metrics of adult bee mass and brood surface area, and measurements of bee head weight, newly-emerged bee weight, and hypopharyngeal gland size were not significantly affected by methoxyfenozide exposure, suggesting that there may be significant effects on honey bee colony behavior and health in the field that are difficult to detect using standard methods for assessing bee colonies and individuals. The second experiment was continued into the following spring, using the same treatment groups as in the fall. Fewer differences were observed among groups in the spring than the fall, possibly because of abundant spring forage and consequent reduced treatment patty consumption. Residue analyses showed that: 1) observed methoxyfenozide concentrations in treatment patty were about 18-60% lower than the calculated concentrations; 2) no residues were observed in wax in any treatment; and 3) methoxyfenozide was detected in bee bread only in the 200 ppb treatment group, at about 1-2.5% of the observed patty concentration.

Using within-day hive weight changes to measure environmental effects on honey bee colonies.

Patterns in within-day hive weight data from two independent datasets in Arizona and California were modeled using piecewise regression, and analyzed with respect to honey bee colony behavior and landscape effects. The regression analysis yielded information on the start and finish of a colony's daily activity cycle, hive weight change at night, hive weight loss due to departing foragers and weight gain due to returning foragers. Assumptions about the meaning of the timing and size of the morning weight changes were tested in a third study by delaying the forager departure times from one to three hours using screen entrance gates. A regression of planned vs. observed departure delays showed that the initial hive weight loss around dawn was largely due to foragers. In a similar experiment in Australia, hive weight loss due to departing foragers in the morning was correlated with net bee traffic (difference between the number of departing bees and the number of arriving bees) and from those data the payload of the arriving bees was estimated to be 0.02 g. The piecewise regression approach was then used to analyze a fifth study involving hives with and without access to natural forage. The analysis showed that, during a commercial pollination event, hives with previous access to forage had a significantly higher rate of weight gain as the foragers returned in the afternoon, and, in the weeks after the pollination event, a significantly higher rate of weight loss in the morning, as foragers departed. This combination of continuous weight data and piecewise regression proved effective in detecting treatment differences in foraging activity that other methods failed to detect.

Monitoring Colony-level Effects of Sublethal Pesticide Exposure on Honey Bees.

The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment methods. Here we describe methods to measure the quantities of adult bees, brood, and food resources by weighing hives and hive parts, by photographing frames, and by installing hives on scales and with internal sensors. Data from these periodic evaluations are then combined with running average and daily detrended data on hive weight and internal hive temperature. The resulting datasets have been used to detect colony-level effects of imidacloprid applied in a sugar syrup as low as 5 parts per billion. The methods are objective, require little training, and provide permanent records in the form of sensor output and photographs.

Sublethal Effects of Imidacloprid on Honey Bee Colony Growth and Activity at Three Sites in the U.S.

Imidacloprid is a neonicotinoid pesticide heavily used by the agricultural industry and shown to have negative impacts on honey bees above certain concentrations. We evaluated the effects of different imidacloprid concentrations in sugar syrup using cage and field studies, and across different environments. Honey bee colonies fed sublethal concentrations of imidicloprid (0, 5, 20 and 100 ppb) over 6 weeks in field trials at a desert site (Arizona), a site near intensive agriculture (Arkansas) and a site with little nearby agriculture but abundant natural forage (Mississippi) were monitored with respect to colony metrics, such as adult bee and brood population sizes, as well as pesticide residues. Hive weight and internal hive temperature were monitored continuously over two trials in Arizona. Colonies fed 100 ppb imidacloprid in Arizona had significantly lower adult bee populations, brood surface areas and average frame weights, and reduced temperature control, compared to colonies in one or more of the other treatment groups, and consumption rates of those colonies were lower compared to other colonies in Arizona and Arkansas, although no differences in capped brood or average frame weight were observed among treatments in Arkansas. At the Mississippi site, also rich in alternative forage, colonies fed 5 ppb imidacloprid had less capped brood than control colonies, but contamination of control colonies was detected. In contrast, significantly higher daily hive weight variability among colonies fed 5 ppb imidacloprid in Arizona suggested greater foraging activity during a nectar flow post treatment, than any other treatment group. Imidacloprid concentrations in stored honey corresponded well with the respective syrup concentrations fed to the colonies and remained stable within the hive for at least 7 months after the end of treatment.

Comparison of productivity of colonies of honey bees, Apis mellifera, supplemented with sucrose or high fructose corn syrup.

Honey bee colony feeding trials were conducted to determine whether differential effects of carbohydrate feeding (sucrose syrup (SS) vs. high fructose corn syrup, or HFCS) could be measured between colonies fed exclusively on these syrups. In one experiment, there was a significant difference in mean wax production between the treatment groups and a significant interaction between time and treatment for the colonies confined in a flight arena. On average, the colonies supplied with SS built 7916.7 cm(2) ± 1015.25 cm(2) honeycomb, while the colonies supplied with HFCS built 4571.63 cm(2) ± 786.45 cm(2). The mean mass of bees supplied with HFCS was 4.65 kg (± 0.97 kg), while those supplied with sucrose had a mean of 8.27 kg (± 1.26). There was no significant difference between treatment groups in terms of brood rearing. Differences in brood production were complicated due to possible nutritional deficiencies experienced by both treatment groups. In the second experiment, colonies supplemented with SS through the winter months at a remote field site exhibited increased spring brood production when compared to colonies fed with HFCS. The differences in adult bee populations were significant, having an overall average of 10.0 ± 1.3 frames of bees fed the sucrose syrup between November 2008 and April 2009, compared to 7.5 ± 1.6 frames of bees fed exclusively on HFCS. For commercial queen beekeepers, feeding the right supplementary carbohydrates could be especially important, given the findings of this study.

Carbohydrate composition of high-fructose corn syrups (HFCS) used for bee feeding: effect on honey composition.

In this study, the carbohydrate composition of high-fructose corn syrups (HFCS) from commercial manufacturers as well as from beekeepers was characterized by GC-MS. Sucrose syrups (SS) were also included in this work for comparison. Fructosyl-fructoses and some unknown carbohydrates, which could correspond to fructosyl-glucoses, have been detected in HFCS for the first time, whereas SS were mainly characterized by the high contents of sucrose. Hydroxymethylfurfural (HMF) content of samples supplied by beekeepers was much more variable; the mean level of HMF was 64.61 ppm (+/-16.92 ppm, 95% CI ranging from 26.91 to 102.31 ppm). Syrups were used to feed caged bees and the resulting honeys produced were analyzed in order to determine their influence in carbohydrate composition. Fructosyl-fructoses were mainly detected in honeys from bees fed with HFCS, but not from those honeys coming from free-flying bees or bees fed with SS.