Honey bees switch mechanisms to drink deep nectar efficiently.

The feeding mechanisms of animals constrain the spectrum of resources that they can exploit profitably. For floral nectar eaters, both corolla depth and nectar properties have marked influence on foraging choices. We report the multiple strategies used by honey bees to efficiently extract nectar at the range of sugar concentrations and corolla depths they face in nature. Honey bees can collect nectar by dipping their hairy tongues or capillary loading when lapping it, or they can attach the tongue to the wall of long corollas and directly suck the nectar along the tongue sides. The honey bee feeding apparatus is unveiled as a multifunctional tool that can switch between lapping and sucking nectar according to the instantaneous ingesting efficiency, which is determined by the interplay of nectar-mouth distance and sugar concentration. These versatile feeding mechanisms allow honey bees to extract nectar efficiently from a wider range of floral resources than previously appreciated and endow them with remarkable adaptability to diverse foraging environments.

Drinking made easier: honey bee tongues dip faster into warmer and/or less viscous artificial nectar.

Optimal concentrations for nectar drinking are limited by the steep increase in the viscosity of sugar solutions with concentration. However, nectar viscosity is inversely related to temperature, which suggests there are advantages to foraging from flowers that are warmer than the surrounding air. The honey bee (Apis mellifera L.) dips nectar using a hairy tongue. However, the microscopic dynamics of the tongue while the bee ingests nectar of varying concentration, viscosity and temperature are unknown. In this study, we found that honey bees respond to the variation of nectar properties by regulating dipping frequency. Through high-speed imaging, we discovered that the honey bee traps warmer sucrose solutions with a quicker tongue. The honey bee dips the warmest and most dilute solution (40°C and 25% w/w sucrose) 1.57 times as fast as the coldest and thickest solution (20°C and 45% w/w sucrose). When the viscosity of different sucrose concentrations was kept constant by adding the inert polysaccharide Tylose, honey bees dipped nectar at constant frequency. We propose a fluid mechanism model to elucidate potential effects on sucrose intake and show that higher dipping frequency can increase the volumetric and energetic intake rates by 125% and 15%, respectively. Our findings broaden insights into how honey bees adapt to foraging constraints from the perspective of tongue dynamics, and demonstrate that elevated intrafloral temperatures and lower nectar viscosity can improve the volumetric and energetic intake rates of pollinators.

Adult diet does not compensate for impact of a poor larval diet on stress resistance in a tephritid fruit fly.

Adult holometabolous insects may derive metabolic resources from either larval or adult feeding, but little is known of whether adult diets can compensate for deficiencies in the larval diet in terms of stress resistance. We investigated how stress resistance is affected and compensated for by diet across life stages in the marula fruit fly Ceratitis cosyra (Diptera: Tephritidae). Larvae were fed diets containing either 8% torula yeast, the standard diet used to rear this species, or 1% yeast (low protein content similar to known host fruit). At emergence, adults from each larval diet were tested for initial mass, water content, body composition, and desiccation and starvation resistance or they were allocated to one of two adult diet treatments: sucrose only, or sucrose and yeast hydrolysate. The same assays were then repeated after 10 days of adult feeding. Development on a low protein larval diet led to lower body mass and improved desiccation and starvation resistance in newly emerged adults, even though adults from the high protein larval diet had the highest water content. Adult feeding decreased desiccation or starvation resistance, regardless of the diet provided. Irrespective of larval diet history, newly emerged, unfed adults had significantly higher dehydration tolerance than those that were fed. Lipid reserves played a role in starvation resistance. There was no evidence for metabolic water from stored nutrients extending desiccation resistance. Our findings show the possibility of a nutrient-poor larval environment leading to correlated improvement in adult performance, at least in the short term.

Nutritional Physiology and Ecology of Honey Bees.

Honey bees feed on floral nectar and pollen that they store in their colonies as honey and bee bread. Social division of labor enables the collection of stores of food that are consumed by within-hive bees that convert stored pollen and honey into royal jelly. Royal jelly and other glandular secretions are the primary food of growing larvae and of the queen but are also fed to other colony members. Research clearly shows that bees regulate their intake, like other animals, around specific proportions of macronutrients. This form of regulation is done as individuals and at the colony level by foragers.

Effect of Brood Pheromone on Survival and Nutrient Intake of African Honey Bees (Apis mellifera scutellata) under Controlled Conditions.

The influence of pheromones on insect physiology and behavior has been thoroughly reported for numerous aspects, such as attraction, gland development, aggregation, mate and kin recognition. Brood pheromone (BP) is released by honey bee larvae to indicate their protein requirements to the colony. Although BP is known to modulate pollen and protein consumption, which in turn can affect physiological and morphological parameters, such as hypopharyngeal gland (HPG) development and ovarian activation, few studies have focused on the effect of BP on nutritional balance. In this study, we exposed newly emerged worker bees for 14 d and found that BP exposure increased protein intake during the first few days, with a peak in consumption at day four following exposure. BP exposure decreased survival of caged honey bees, but did not affect either the size of the HPG acini or ovarian activation stage. The uncoupling of the BP releaser effect, facilitated by working under controlled conditions, and the presence of larvae as stimulating cues are discussed.

Do sunbirds use taste to decide how much to drink?

Nectarivorous birds typically consume smaller meals of more concentrated than of less concentrated sugar solutions. It is not clear, however, whether they use taste to decide how much to consume or whether they base this decision on post-ingestive feedback. Taste, a cue to nectar concentration, is available to nectarivores during ingestion whereas post-ingestive information about resource quality becomes available only after a meal. When conditions are variable, we would expect nectarivorous birds to base their decisions on how much to consume on taste, as post-ingestive feedback from previous meals would not be a reliable cue to current resource quality. Here, we tested whether white-bellied sunbirds (Cinnyris talatala), foraging from an array of artificial flowers, use taste to decide how much to consume per meal when nectar concentration is highly variable: they did not. Instead, how much they chose to consume per meal appeared to depend on the energy intake at the previous meal, that is how hungry they were. Our birds did, however, appear to use taste to decide how much to consume per flower visited within a meal. Unexpectedly, some individuals preferred to consume more from flowers with lower concentration rewards and some preferred to do the opposite. We draw attention to the fact that many studies perhaps misleadingly claim that birds use sweet taste to inform their foraging decisions, as they analyse mean data for multiple meals over which post-ingestive feedback will have become available rather than data for individual meals when only sensory information is available. We discuss how conflicting foraging rules could explain why sunbirds do not use sweet taste to inform their meal size decisions.

Nutrient balancing of the adult worker bumblebee (Bombus terrestris) depends on the dietary source of essential amino acids.

Animals carefully regulate the amount of protein that they consume. The quantity of individual essential amino acids (EAAs) obtained from dietary protein depends on the protein source, but how the proportion of EAAs in the diet affects nutrient balancing has rarely been studied. Recent research using the Geometric Framework for Nutrition has revealed that forager honeybees who receive much of their dietary EAAs from floral nectar and not from solid protein have relatively low requirements for dietary EAAs. Here, we examined the nutritional requirements for protein and carbohydrates of foragers of the buff-tailed bumblebee Bombus terrestris. By using protein (sodium caseinate) or an equimolar mixture of the 10 EAAs, we found that the intake target (nutritional optimum) of adult workers depended on the source and proportion of dietary EAAs. When bees consumed caseinate-containing diets in a range of ratios between 1:250 and 1:25 (protein to carbohydrate), they achieved an intake target (IT) of 1:149 (w/w). In contrast to those fed protein, bees fed the EAA diets had an IT more biased towards carbohydrates (1:560 w/w) but also had a greater risk of death than those fed caseinate. We also tested how the dietary source of EAAs affected free AAs in bee haemolymph. Bees fed diets near their IT had similar haemolymph AA profiles, whereas bees fed diets high in caseinate had elevated levels of leucine, threonine, valine and alanine in the haemolymph. We found that like honeybees, bumblebee workers prioritize carbohydrate intake and have a relatively low requirement for protein. The dietary source of EAAs influenced both the ratio of protein/EAA to carbohydrate and the overall amount of carbohydrate eaten. Our data support the idea that EAAs and carbohydrates in haemolymph are important determinants of nutritional state in insects.

Nutritional balance of essential amino acids and carbohydrates of the adult worker honeybee depends on age.

Dietary sources of essential amino acids (EAAs) are used for growth, somatic maintenance and reproduction. Eusocial insect workers such as honeybees are sterile, and unlike other animals, their nutritional needs should be largely dictated by somatic demands that arise from their role within the colony. Here, we investigated the extent to which the dietary requirements of adult worker honeybees for EAAs and carbohydrates are affected by behavioural caste using the Geometric Framework for nutrition. The nutritional optimum, or intake target (IT), was determined by confining cohorts of 20 young bees or foragers to liquid diets composed of specific proportions of EAAs and sucrose. The IT of young, queenless bees shifted from a proportion of EAAs-to-carbohydrates (EAA:C) of 1:50 towards 1:75 over a 2-week period, accompanied by a reduced lifespan on diets high in EAAs. Foragers required a diet high in carbohydrates (1:250) and also had low survival on diets high in EAA. Workers exposed to queen mandibular pheromone lived longer on diets high in EAA, even when those diets contained 5× their dietary requirements. Our data show that worker honeybees prioritize their intake of carbohydrates over dietary EAAs, even when overeating EAAs to obtain sufficient carbohydrates results in a shorter lifespan. Thus, our data demonstrate that even when young bees are not nursing brood and foragers are not flying, their nutritional needs shift towards a diet largely composed of carbohydrates when they make the transition from within-hive duties to foraging.

Drinking problems on a 'simple' diet: physiological convergence in nectar-feeding birds.

Regulation of energy and water are by necessity closely linked in avian nectarivores, because the easily available sugars in nectar are accompanied by an excess of water but few electrolytes. In general, there is convergence in morphology and physiology between three main lineages of avian nectarivores that have evolved on different continents - the hummingbirds, sunbirds and honeyeaters. These birds show similar dependence of sugar preferences on nectar concentration, high intestinal sucrase activity and rapid absorption of hexoses via mediated and paracellular routes. There are differences, however, in how these lineages deal with energy challenges, as well as processing the large volumes of preformed water ingested in nectar. While hummingbirds rely on varying renal water reabsorption, the passerine nectarivores modulate intestinal water absorption during water loading, thus reducing the impact on the kidneys. Hummingbirds do not generally cope with salt loading, and have renal morphology consistent with their ability to produce copious dilute urine; by contrast, as well as being able to deal with dilute diets, honeyeaters and sunbirds are more than capable of dealing with moderately high levels of added electrolytes. And finally, in response to energy challenge, hummingbirds readily resort to torpor, while the passerines show renal and digestive responses that allow them to deal with short-term fasts and rapidly restore energy balance without using torpor. In conclusion, sunbirds and honeyeaters demonstrate a degree of physiological plasticity in dealing with digestive and renal challenges of their nectar diet, while hummingbirds appear to be more constrained by this diet.

Honeybees prefer warmer nectar and less viscous nectar, regardless of sugar concentration.

The internal temperature of flowers may be higher than air temperature, and warmer nectar could offer energetic advantages for honeybee thermoregulation, as well as being easier to drink owing to its lower viscosity. We investigated the responses of Apis mellifera scutellata (10 colonies) to warmed 10% w/w sucrose solutions, maintained at 20-35°C, independent of low air temperatures, and to 20% w/w sucrose solutions with the viscosity increased by the addition of the inert polysaccharide Tylose (up to the equivalent of 34.5% sucrose). Honeybee crop loads increased with nectar temperature, as did the total consumption of sucrose solutions over 2 h by all bees visiting the feeders. In addition, the preference of marked honeybees shifted towards higher nectar temperatures with successive feeder visits. Crop loads were inversely proportional to the viscosity of the artificial nectar, as was the total consumption of sucrose solutions over 2 h. Marked honeybees avoided higher nectar viscosities with successive feeder visits. Bees thus showed strong preferences for both warmer and less viscous nectar, independent of changes in its sugar concentration. Bees may benefit from foraging on nectars that are warmer than air temperature for two reasons that are not mutually exclusive: reduced thermoregulatory costs and faster ingestion times due to the lower viscosity.

Gastrointestinal and renal responses to variable water intake in whitebellied sunbirds and New Holland honeyeaters.

Nectarivores face a constant challenge in terms of water balance, experiencing water loading or dehydration when switching between food plants or between feeding and fasting. To understand how whitebellied sunbirds and New Holland honeyeaters meet the challenges of varying preformed water load, we used the elimination of intramuscular-injected [(14)C]-l-glucose and (3)H2O to quantify intestinal and renal water handling on diets varying in sugar concentration. Both sunbirds and honeyeaters showed significant modulation of intestinal water absorption, allowing excess water to be shunted through the intestine when on dilute diets. Despite reducing their fractional water absorption, both species showed linear increases in water flux and fractional body water turnover as water intake increased (both afternoon and morning), suggesting that the modulation of fractional water absorption was not sufficient to completely offset dietary water loads. In both species, glomerular filtration rate was independent of water gain (but was higher for the afternoon), as was renal fractional water reabsorption (measured in the afternoon). During the natural overnight fast, both sunbirds and honeyeaters arrested whole kidney function. Evaporative water loss in sunbirds was variable but correlated with water gain. Both sunbirds and honeyeaters appear to modulate intestinal water absorption as an important component of water regulation to help deal with massive preformed water loads. Shutting down glomerular filtration rate during the overnight fast is another way of saving energy for osmoregulatory function. Birds maintain osmotic balance on diets varying markedly in preformed water load by varying both intestinal water absorption and excretion through the intestine and kidneys.

Selection on female reproductive schedules in the marula fly, Ceratitis cosyra (Diptera: Tephritidae) affects dietary optima for female reproductive traits but not lifespan.

Introduction: A changing environment can select on life-history traits and trade-offs in a myriad of ways. For example, global warming may shift phenology and thus the availability of host-plants. This may alter selection on survival and fertility schedules in herbivorous insects. If selection on life-histories changes, this may in turn select for altered nutrient intake, because the blend of nutrients organisms consume helps determine the expression of life-history traits. However, we lack empirical work testing whether shifts in the timing of oviposition alter nutrient intake and life-history strategies. Methods: We tested in the marula fruit fly, Ceratitis cosyra, how upward-selection on the age of female oviposition, in comparison with laboratory adapted control flies, affects the sex-specific relationship between protein and carbohydrate intake and life-history traits including lifespan, female lifetime egg production and daily egg production. We then determined the macronutrient ratio consumed when flies from each selection line and sex were allowed to self-regulate their intake. Results: Lifespan, lifetime egg production and daily egg production were optimised at similar protein to carbohydrate (P:C) ratios in flies from both selection lines. Likewise, females and males of both lines actively defended similar nutrient intake ratios (control =1:3.6 P:C; upward-selected = 1:3.2 P:C). Discussion: Our results are comparable to those in non-selected C. cosyra, where the optima for each trait and the self-selected protein to carbohydrate ratio observed were nearly identical. The nutrient blend that needs to be ingested for optimal expression of a given trait appeared to be well conserved across laboratory adapted and experimentally selected populations. These results suggest that in C. cosyra, nutritional requirements do not respond to a temporal change in oviposition substrate availability.

Sweet solutions: nectar chemistry and quality.

Nectar, the main floral reward for pollinators, varies greatly in composition and concentration. The assumption that nectar quality is equivalent to its sugar (energy) concentration is too simple. Diverse non-sugar components, especially amino acids and secondary metabolites, play various roles in nutrition and health of pollinators. Many nectar compounds have indirect effects by altering the foraging behaviour of pollinators or protecting them from disease. This review also emphasizes the water component of nectar, often ignored because of evaporative losses and difficulties in sampling small nectar volumes. Nectar properties vary with environmental factors, pollinator visits and microbial contamination. Pollination mutualisms depend on the ability of insect and vertebrate pollinators to cope with and benefit from the variation and diversity in nectar chemistry. This article is part of the theme issue 'Natural processes influencing pollinator health: from chemistry to landscapes'.

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.

Oxidative Damage Is Influenced by Diet But Unaffected by Selection for Early Age of Oviposition in the Marula Fly, Ceratitis cosyra (Diptera: Tephritidae).

The expression of life-history traits, such as lifespan or reproductive effort, is tightly correlated with the amount and blend of macronutrients that individuals consume. In a range of herbivorous insects, consuming high protein to carbohydrate ratios (P:C) decreases lifespan but increases female fecundity. In other words, females face a resource-based trade-off between lifespan and fecundity. Redox metabolism may help mediate this trade-off, if oxidative damage is elevated by reproductive investment and if this damage, in turn, reduces lifespan. Here, we test how diets varying in P:C ratio affect oxidative damage and antioxidant protection in female and male of the marula fly, Ceratitis cosyra (Diptera: Tephritidae). We use replicated lines that have been subjected to experimental evolution and differ in their lifespan and reproductive scheduling. We predicted that high fecundity would be associated with high oxidative damage and reduced antioxidant defences, while longer lived flies would show reduced damage and elevated antioxidant defences. However, higher levels of oxidative damage were observed in long-lived control lines than selection lines, but only when fed the diet promoting lifespan. Flies fed diets promoting female fecundity (1:4 and 1:2 P:C) suffered greater oxidative damage to lipids than flies fed the best diet (0:1 P:C) for lifespan. Total antioxidant capacity was not affected by the selection regime or nutrition. Our results reiterate the importance of nutrition in affecting life-history traits, but suggest that in C. cosyra, reactive oxygen species play a minimal role in mediating dietary trade-offs between lifespan and reproduction.

Natural processes influencing pollinator health.

Evidence from the last few decades indicates that pollinator abundance and diversity are at risk, with many species in decline. Anthropogenic impacts have been the focus of much recent work on the causes of these declines. However, natural processes, from plant chemistry, nutrition and microbial associations to landscape and habitat change, can also profoundly influence pollinator health. Here, we argue that these natural processes require greater attention and may even provide solutions to the deteriorating outlook for pollinators. Existing studies also focus on the decline of individuals and colonies and only occasionally at population levels. In the light of this we redefine pollinator health and argue that a top-down approach is required focusing at the ecological level of communities. We use examples from the primary research, opinion and review articles published in this special issue to illustrate how natural processes influence pollinator health, from community to individuals, and highlight where some of these processes could mitigate the challenges of anthropogenic and natural drivers of change. This article is part of the theme issue 'Natural processes influencing pollinator health: from chemistry to landscapes'.

Experimental Manipulation of Air Temperature in Captivity Appears Unsuitable for Evaluating Fecal Glucocorticoid Metabolite Responses of Wild-Caught Birds to Heat Exposure.

AbstractNoninvasive measurement of stress-related alterations in fecal glucocorticoid metabolite (fGCM) concentrations has considerable potential for quantifying physiological responses to very hot weather in free-ranging birds, but practical considerations related to sampling will often make this method feasible only for habituated study populations. Here we evaluate an alternate approach, the use of experimentally manipulated thermal environments for evaluating stress responses to high environmental temperatures in wild-caught birds housed in captivity. Using an enzyme immunoassay utilizing antibodies against 5ß-pregnane-3α,11ß,21-triol-20-one-CMO∶BSA (tetrahydrocorticosterone), we quantified fGCMs in captive individuals of three southern African arid-zone species (southern pied babblers [Turdoides bicolor], white-browed sparrow-weavers [Plocepasser mahali], and southern yellow-billed hornbills [Tockus leucomelas]) experiencing daily air temperature maxima (Tmax) ranging from 30°-32°C to 42°-44°C. For none of the three species did Tmax emerge as a significant predictor of elevated fGCM concentrations, and no stress response to simulated hot weather was evident. The apparent lack of a stress response to Tmax = 42°C in captive southern pied babblers contrasts with linear increases in fGCMs at Tmax > 38°C in free-ranging conspecifics. The lack of an effect of Tmax on fGCM levels may potentially be explained by several factors, including differences in operative temperatures and the availability of water and food between free-ranging and captive settings or the stress effect of captivity itself. Our results suggest that experimental manipulations of thermal environments experienced by wild-caught captive birds have limited usefulness for testing hypotheses concerning the effects of hot weather events on fGCM (and, by extension, glucocorticoid) concentrations.

Sucrose digestion capacity in birds shows convergent coevolution with nectar composition across continents.

The major lineages of nectar-feeding birds (hummingbirds, sunbirds, honeyeaters, flowerpiercers, and lorikeets) are considered examples of convergent evolution. We compared sucrose digestion capacity and sucrase enzymatic activity per unit intestinal surface area among 50 avian species from the New World, Africa, and Australia, including 20 nectarivores. With some exceptions, nectarivores had smaller intestinal surfaces, higher sucrose hydrolysis capacity, and greater sucrase activity per unit intestinal area. Convergence analysis showed high values for sucrose hydrolysis and sucrase activity per unit intestinal surface area in specialist nectarivores, matching the high proportion of sucrose in the nectar of the plants they pollinate. Plants pollinated by generalist nectar-feeding birds in the Old and New Worlds secrete nectar in which glucose and fructose are the dominant sugars. Matching intestinal enzyme activity in birds and nectar composition in flowers appears to be an example of convergent coevolution between plants and pollinators on an intercontinental scale.

Dilute bird nectars: viscosity constrains food intake by licking in a sunbird.

Floral nectars of bird-pollinated plants are relatively dilute. One hypothesis proposed to explain this concerns the difficulty for birds of drinking nectar of high viscosity. We examined the effects of viscosity, separately from those of sugar concentration, on feeding by captive whitebellied sunbirds (Cinnyris talatala). Viscosities of artificial nectar (sucrose solutions ranging in concentration from 0.25 to 1.5 mol/l) were altered with Tylose, an inert polysaccharide. Food consumption was measured over 3 h, and lick frequency and duration were recorded using photodetection devices on feeding apertures too small for the bill but large enough for the extended tongue. Volumetric intake rates (ml/s) were inversely proportional to nectar viscosity, and were similar over the range of sucrose concentrations when viscosity was held constant. Sucrose intake rates (mg/s) remained the same on pure sucrose solutions, but they decreased with increasing viscosity at a constant sucrose concentration. Lick frequencies and tongue loads were reduced at high viscosities, and lick duration increased, which confirms that sunbirds take longer to ingest viscous solutions. Licking behavior was remarkably similar in birds feeding on different sucrose concentrations if viscosity was held constant. Nectar ingestion rate is determined by viscosity; however, total food intake is mainly modulated by sugar concentration. Similar effects of food viscosity have been observed in insects that suck nectar.

Convergence of carbohydrate-biased intake targets in caged worker honeybees fed different protein sources.

The nutritional needs of bees are supplied by nectar carbohydrates and by protein and other nutrients in pollen but little is known of how bees achieve nutritional balance. Using newly emerged caged worker honeybees (Apis mellifera scutellata), we investigated whether bees maintain their intake target when confined to pairs of imbalanced complementary diets varying in protein to carbohydrate (P:C) ratio. Diets were formulated using three protein sources [casein, royal jelly or Feed-Bee (a natural pollen substitute)] and sucrose. Within each protein type, honeybees switched between complementary diets and converged on the same P:C intake target. However, this target differed between protein types: P:C ratios were 1:12, 1:14 and 1:11 on casein, royal jelly and Feed-Bee diets, respectively. Except for an early peak in protein consumption on royal jelly diets, these strongly convergent ratios remained constant over the 14 day experiment. This is probably due to the absence of brood, reflected in relatively stable values measured for haemolymph protein concentration and hypopharyngeal gland activation in bees on Feed-Bee diets. Performance of caged workers was also assessed in terms of survival and ovarian activation. Survival was highest on casein diets and lowest on Feed-Bee diets but ovarian activation was highest on royal jelly diets and lowest on casein diets. This may be due to additional components in Feed-Bee and royal jelly (e.g. fatty acids), which are needed to activate the ovaries but also reduce survival. Nutrient intake of broodless workers is directly related to their own physiological requirements, and the strong carbohydrate bias may reflect the high metabolic rate of honeybees even under resting conditions.