Honey bees save energy in honey processing by dehydrating nectar before returning to the nest.

Honey bees process nectar into honey by active evaporation on the tongue and passive evaporation involving nest ventilation and fanning behaviour, as well as enzymatic action. The elimination of excess water from nectar carries considerable energetic costs. The concentration of the nectar load is assumed to remain constant during transport. However, some of this water elimination may occur before foragers return to the nest and pass their nectar loads to receiver bees. In honey bees captured while foraging in Macadamia orchards, we show that the nectar in their crops has approximately twice the sugar concentration of the fresh nectar in flowers. This was true for four Macadamia cultivars, with up to 75% of the initial water content being removed. There is a further concentration increase in the crops of returning bees captured at the hive entrance. The only possible route of water elimination from the crop is via evaporation from the mouthparts. We calculate the savings in honey processing costs to be on average 35 times more than the reduction in flight costs due to reduced body mass. Pre-concentration of nectar in foraging honey bees may be widespread, and of crucial importance for honey storage.

Drivers of colony losses.

Over the past decade, in some regions of the world, honey bee (Apis mellifera L.) colonies have experienced rates of colony loss that are difficult for beekeepers to sustain. The reasons for losses are complex and interacting, with major drivers including Varroaand related viruses, pesticides, nutrition and beekeeper practices. In these endeavors it has also become apparent that defining a dead colony, and singling out the effects of specific drivers of loss, is not so straightforward. Using the class of neonicotinoid pesticides as an example we explain why quantifying risk factor impact at the colony level is at times elusive and in some cases unpractical. In this review, we discuss the caveats of defining and quantifying dead colonies. We also summarize the current leading drivers of colony losses, their interactions and the most recent research on their effects on colony mortality.

Neonicotinoids decrease sucrose responsiveness of honey bees at first contact.

For two decades, neonicotinoid insecticides have been extensively used worldwide. Targeting neuronal receptors, they have deleterious effects on the behaviour and physiology of many insects. Bees are exposed to these insecticides in pollen and nectar while providing pollination services to agricultural crops, and neonicotinoids have been shown to impair navigation and decrease their foraging activity. We have previously reported the effect of dietary thiamethoxam on sucrose responsiveness of young worker bees. Here, we exposed caged foragers to sublethal acute doses of clothianidin, imidacloprid, and thiamethoxam, then tested them individually for sucrose responsiveness using standard methods. In addition, we tested the response to a range of sucrose solutions laced with neonicotinoids on bees previously unexposed to neonicotinoids. This paradigm mimics the situation where foragers would first encounter poisoned nectars varying in sugar concentration. Bees were exposed to the insecticides in the feeding solution for 24 h before testing, or in the test solutions, or both. The three compounds had a detrimental effect on responses to mid-to-high sucrose concentrations under all experimental conditions, and unexposed bees tested with laced sucrose displayed unexpected low responses to the higher sucrose concentrations tested. This attenuation of sucrose response is further evidence that neonicotinoids are multisensory disruptors, with potent actions against pollinators and other beneficial insects at first contact.

Digestibility and nutritional value of fresh and stored pollen for honey bees (Apis mellifera scutellata).

Pollen, the main protein source for honey bees, is mixed with regurgitated nectar or honey during collection and then stored as 'bee bread' before its consumption, mainly by young nurse workers. It has been suggested that storage of pollen improves its nutritional value and digestibility, but there is little evidence for such changes. We fed two fresh pollen types of different protein content (aloe and sunflower), and two stored pollen types (sunflower and a mixed pollen), to young caged worker bees. We measured daily consumption of pollen and sucrose solution, and survival after 14 days. At day 14 we recorded ovarian activation and extraction efficiency, by counting empty pollen grains in the rectal contents. Extraction efficiency is a measure of pollen digestibility. Contrary to our predictions, bees did not consume more fresh sunflower pollen than fresh aloe pollen to compensate for the lower protein content of sunflower pollen. In addition, they did not consume less sucrose solution when fed stored pollen diets that are already enriched in sugar. Consumption of stored sunflower pollen resulted in a low protein to carbohydrate (P:C) intake. Survival and ovarian activation were higher on diets giving higher P:C intakes. Extraction efficiency was high (up to 99%) for all pollen diets, and comparison of fresh and stored sunflower pollen showed that storage did not make it easier to digest. Changes to pollen during storage do not confer obvious benefits to honey bees.

Proteomic and metabolomic analysis reveals rapid and extensive nicotine detoxification ability in honey bee larvae.

Despite potential links between pesticides and bee declines, toxicology information on honey bee larvae (Apis mellifera) is scarce and detoxification mechanisms in this development stage are virtually unknown. Larvae are exposed to natural and synthetic toxins present in pollen and nectar through consumption of brood food. Due to the characteristic intensive brood care displayed by honey bees, which includes progressive feeding throughout larval development, it is generally assumed that larvae rely on adults to detoxify for them and exhibit a diminished detoxification ability. We found the opposite. We examined the proteomic and metabolomic responses of in vitro reared larvae fed nicotine (an alkaloid found in nectar and pollen) to understand how larvae cope on a metabolic level with dietary toxins. Larvae were able to effectively detoxify nicotine through an inducible detoxification mechanism. A coordinated stress response complemented the detoxification processes, and we detected significant enrichment of proteins functioning in energy and carbohydrate metabolism, as well as in development pathways, suggesting that nicotine may promote larval growth. Further exploration of the metabolic fate of nicotine using targeted mass spectrometry analysis demonstrated that, as in adult bees, formation of 4-hydroxy-4-(3-pyridyl) butanoic acid, the result of 2'C-oxidation of nicotine, is quantitatively the most significant pathway of nicotine metabolism. We provide conclusive evidence that larvae are capable of effectively catabolising a dietary toxin, suggesting that increased larval sensitivity to specific toxins is not due to diminished detoxification abilities. These findings broaden the current understanding of detoxification biochemistry at different organizational levels in the colony, bringing us closer to understanding the capacity of the colony as a superorganism to tolerate and resist toxic compounds, including pesticides, in the environment.

Effects of a neonicotinoid pesticide on thermoregulation of African honey bees (Apis mellifera scutellata).

Thiamethoxam is a widely used neonicotinoid pesticide that, as agonist of the nicotinic acetylcholine receptors, has been shown to elicit a variety of sublethal effects in honey bees. However, information concerning neonicotinoid effects on honey bee thermoregulation is lacking. Thermoregulation is an essential ability for the honey bee that guarantees the success of foraging and many in-hive tasks, especially brood rearing. We tested the effects of acute exposure to thiamethoxam (0.2, 1, 2ng/bee) on the thorax temperatures of foragers exposed to low (22°C) and high (33°C) temperature environments. Thiamethoxam significantly altered honey bee thorax temperature at all doses tested; the effects elicited varied depending on the environmental temperature and pesticide dose to which individuals were exposed. When bees were exposed to the high temperature environment, the high dose of thiamethoxam increased their thorax temperature 1-2h after exposure. When bees were exposed to the low temperature, the higher doses of the neonicotinoid reduced bee thorax temperatures 60-90min after treatment. In both experiments, the neonicotinoid decreased the temperature of bees the day following the exposure. After a cold shock (5min at 4°C), the two higher doses elicited a decrease of the thorax temperature, while the lower dose caused an increase, compared to the control. These alterations in thermoregulation caused by thiamethoxam may affect bee foraging activity and a variety of in-hive tasks, likely leading to negative consequences at the colony level. Our results shed light on sublethal effect of pesticides which our bees have to deal with.

Sucrose Sensitivity of Honey Bees Is Differently Affected by Dietary Protein and a Neonicotinoid Pesticide.

Over a decade, declines in honey bee colonies have raised worldwide concerns. Several potentially contributing factors have been investigated, e.g. parasites, diseases, and pesticides. Neonicotinoid pesticides have received much attention due to their intensive use in crop protection, and their adverse effects on many levels of honey bee physiology led the European Union to ban these compounds. Due to their neuronal target, a receptor expressed throughout the insect nervous system, studies have focused mainly on neuroscience and behaviour. Through the Geometric Framework of nutrition, we investigated effects of the neonicotinoid thiamethoxam on survival, food consumption and sucrose sensitivity of honey bees (Apis mellifera). Thiamethoxam did not affect protein and carbohydrate intake, but decreased responses to high concentrations of sucrose. Interestingly, when bees ate fixed unbalanced diets, dietary protein facilitated better sucrose detection. Both thiamethoxam and dietary protein influenced survival. These findings suggest that, in the presence of a pesticide and unbalanced food, honey bee health may be severely challenged. Consequences for foraging efficiency and colony activity, cornerstones of honey bee health, are also discussed.

Resistance rather than tolerance explains survival of savannah honeybees (Apis mellifera scutellata) to infestation by the parasitic mite Varroa destructor.

Varroa destructor is considered the most damaging parasite affecting honeybees (Apis mellifera L.). However, some honeybee populations such as the savannah honeybee (Apis mellifera scutellata) can survive mite infestation without treatment. It is unclear if survival is due to resistance mechanisms decreasing parasite reproduction or to tolerance mechanisms decreasing the detrimental effects of mites on the host. This study investigates both aspects by quantifying the reproductive output of V. destructor and its physiological costs at the individual host level. Costs measured were not consistently lower when compared with susceptible honeybee populations, indicating a lack of tolerance. In contrast, reproduction of V. destructor mites was distinctly lower than in susceptible populations. There was higher proportion of infertile individuals and the reproductive success of fertile mites was lower than measured to date, even in surviving populations. Our results suggest that survival of savannah honeybees is based on resistance rather than tolerance to this parasite. We identified traits that may be useful for breeding programmes aimed at increasing the survival of susceptible populations. African honeybees may have benefited from a lack of human interference, allowing natural selection to shape a population of honeybees that is more resistant to Varroa mite infestation.

Impact of Varroa destructor on honeybee (Apis mellifera scutellata) colony development in South Africa.

The devastating effects of Varroa destructor Anderson & Trueman on European honeybee colonies (Apis mellifera L.) have been well documented. Not only do these mites cause physical damage to parasitised individuals when they feed on them, they also transmit viruses and other pathogens, weaken colonies and can ultimately cause their death. Nevertheless, not all honeybee colonies are doomed once Varroa mites become established. Some populations, such as the savannah honeybee, A. m. scutellata, have become tolerant after the introduction of the parasite and are able to withstand the presence of these mites without the need for acaricides. In this study, we measured daily Varroa mite fall, Varroa infestation rates of adult honeybees and worker brood, and total Varroa population size in acaricide treated and untreated honeybee colonies. In addition, honeybee colony development was compared between these groups in order to measure the cost incurred by Varroa mites to their hosts. Daily Varroa mite fall decreased over the experimental period with different dynamics in treated and untreated colonies. Varroa infestation rates in treated adult honeybees and brood were lower than in untreated colonies, but not significantly so. Thus, indicating a minimal benefit of treatment thereby suggesting that A. m. scutellata have the ability to maintain mite populations at low levels. We obtained baseline data on Varroa population dynamics in a tolerant honeybee over the winter period. Varroa mites appeared to have a low impact on this honeybee population, given that colony development was similar in the treated and untreated colonies.

Seasonal prevalence of pathogens and parasites in the savannah honeybee (Apis mellifera scutellata).

The loss of Apis mellifera L. colonies in recent years has, in many regions of the world, been alarmingly high. No single cause has been identified for these losses, but the interactions between several factors (mostly pathogens and parasites) have been held responsible. Work in the Americas on honeybees originating mainly from South Africa indicates that Africanised honeybees are less affected by the interplay of pathogens and parasites. However, little is known about the health status of South African honeybees (A. m. scutellata and A. m. capensis) in relation to pathogens and parasites. We therefore compared the seasonal prevalence of honeybee pathogens (viruses, bacteria, fungi) and parasites (mites, bee lice, wax moth, small hive beetles, A. m. capensis social parasites) between sedentary and migratory A. m. scutellata apiaries situated in the Gauteng region of South Africa. No significant differences were found in the prevalence of pathogens and parasites between sedentary and migratory apiaries. Three (Black queen cell virus, Varroa destructor virus 1 and Israeli acute paralysis virus) of the eight viruses screened were detected, a remarkable difference compared to European honeybees. Even though no bacterial pathogens were detected, Nosema apis and Chalkbrood were confirmed. All of the honeybee parasites were found in the majority of the apiaries with the most common parasite being the Varroa mite. In spite of hosting few pathogens, yet most parasites, A. m. scutellata colonies appeared to be healthy.

Bees get a head start on honey production.

Nectar concentration is assumed to remain constant during transport by honeybees between flowers and hive. We sampled crop contents of nectar foragers on Aloe greatheadii var. davyana, a major winter bee plant in South Africa. The nectar is dilute (approx. 20% w/w), but the crop contents of bees captured on flowers are significantly more concentrated. In returning foragers, the concentration increases further to 38-40%, accompanied by a volume decrease. The doubling of sugar concentration suggests that nectar is regurgitated onto the tongue and evaporated during foraging and on the return flight. Processing of the dilute nectar into honey thus begins early, aided by low ambient humidities. This has implications for honeybee thermoregulation, water balance and energetics during foraging, and for the communication of nectar quality to recruits.

Nutritional content of fresh, bee-collected and stored pollen of Aloe greatheadii var. davyana (Asphodelaceae).

Aloe greatheadii var. davyana is the most important indigenous South African bee plant. Fresh, bee-collected and stored pollen of this aloe was collected and analysed for its nutritional content, including amino acid and fatty acid composition. Highly significant differences were found between the three types of pollen. Collection and storage by the bees resulted in increased water (13-21% wet weight) and carbohydrate content (35-61% dry weight), with a resultant decrease in crude protein (51-28% dry weight) and lipid content (10-8% dry weight). Essential amino acids were present in equal or higher amounts than the required minimum levels for honeybee development, with the exception of tryptophan. Fatty acids comprised a higher proportion of total lipid in fresh pollen than in bee-collected and stored pollen. This study is the first to compare the changes that occur in pollen of a single species after collection by honeybees.

Do honeybees, Apis mellifera scutellata, regulate humidity in their nest?

Honeybees are highly efficient at regulating the biophysical parameters of their hive according to colony needs. Thermoregulation has been the most extensively studied aspect of nest homeostasis. In contrast, little is known about how humidity is regulated in beehives, if at all. Although high humidity is necessary for brood development, regulation of this parameter by honeybee workers has not yet been demonstrated. In the past, humidity was measured too crudely for a regulation mechanism to be identified. We reassess this issue, using miniaturised data loggers that allow humidity measurements in natural situations and at several places in the nest. We present evidence that workers influence humidity in the hive. However, there are constraints on potential regulation mechanisms because humidity optima may vary in different locations of the nest. Humidity could also depend on variable external factors, such as water availability, which further impair the regulation. Moreover, there are trade-offs with the regulation of temperature and respiratory gas exchanges that can disrupt the establishment of optimal humidity levels. As a result, we argue that workers can only adjust humidity within sub-optimal limits.