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International audience; AbstractNutritional stress due to habitat transformation and loss is one of several factors contributing to current declines in global bee populations. Bees obtain protein from pollen, which in honeybees is consumed and digested by nurse bees. They then distribute the protein to the rest of the colony in the form of hypopharyngeal gland secretions. Little is known of how efficiently honeybees digest protein. Moreover, antibiotics are used by beekeepers as in-hive treatments for diseases and may interfere with microbial contributions to protein digestion. Caged, newly emerged workers of Apis mellifera scutellata were fed caseinate as protein source, to investigate the effects of protein intake and antibiotic treatment on digestive efficiency. These workers were fed protein:carbohydrate ratios of 1:120, 1:50 and 1:15 or pure sucrose for 9 days. Half the cages received dietary oxytetracycline at a concentration used by beekeepers. Antibiotic exposure did not affect survival or protein consumption. Protein digestive efficiency increased with increasing levels of protein in the diet, although a decrease would have contributed to maintaining nutrient balance. Importantly, we show that antibiotic exposure impaired protein digestive efficiency, especially on low-protein diets. This may be particularly important when colonies are restricted to a single protein deficient source of pollen.

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.

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.

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.

SUMMARYVarroa 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.

The effects of pesticides on honeybee larvae are less understood than for adult bees, even though larvae are chronically exposed to pesticide residues that accumulate in comb and food stores in the hive. We investigated how exposure to a plant alkaloid, nicotine, affects survival, growth and body composition of honeybee larvae. Larvae of Apis mellifera scutellata were reared in vitro and fed throughout development on standard diets with nicotine included at concentrations from 0 to 1000μg/100g diet. Overall mortality across all nicotine treatments was low, averaging 9.8% at the prepupal stage and 18.1% at the white-eyed pupal stage, but survival was significantly reduced by nicotine. The mass of prepupae and white-eyed pupae was not affected by nicotine. In terms of body composition, nicotine affected water content but did not influence either protein or lipid stores of white-eyed pupae. We attribute the absence of consistent negative effects of dietary nicotine to detoxification mechanisms in developing honeybees, which enable them to resist both natural and synthetic xenobiotics.

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.

SummaryThis study reports honey bee, Apis mellifera L., colony losses that occurred in South Africa over two consecutive years. The total losses were 29.6% (95% CI: 22.8–37.5) in 2009–2010 and 46.2% (95% CI: 37.3–55.0) in 2010–2011. Furthermore, the study shows that the capensss worker social parasite, a problem unique to southern Africa, is the main perceived cause, and could explain the significant differences in the number of losses between beekeepers using the subspecies A. m. scutellata and those using the subspecies A. m. capensis. In contrast to previous studies in North America and Europe, we find a significant negative effect of migratory beekeeping practices on the extent of colony losses. Migratory beekeepers lost on average more colonies (35.5% (95% CI 29.7–47.2)) than did stationary beekeepers (17.2% (95% CI 11.2–22.3)). This was especially pronounced when the beekeepers were migrating for the pollination of apples/cherries, eucalyptus, onions and/or sunflowers. The major beekeeper-perceived ...

Summary American foulbrood is one of the most devastating diseases of the honey bee. It is caused by the spore-forming, Gram-positive rod-shaped bacterium Paenibacillus larvae. The recent updated genome assembly and annotation for this pathogen now permits in-depth molecular studies. In this paper, selected techniques and protocols for American foulbrood research are provided, mostly in a recipe-like format that permits easy implementation in the laboratory. Topics covered include: working with Paenibacillus larvae, basic microbiological techniques, experimental infection, and “’omics” and other sophisticated techniques. Further, this chapter covers other technical information including biosafety measures to guarantee the safe handling of this pathogen.

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, 2 ng/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 1–2 h after exposure. When bees were exposed to the low temperature, the higher doses of the neonicotinoid reduced bee thorax temperatures 60–90 min after treatment. In both experiments, the neonicotinoid decreased the temperature of bees the day following the exposure. After a cold shock (5 min 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.

International audience; AbstractHoneybee (Apis mellifera L.) pathogens and parasites and the negative effects thereof on honeybee populations remain an issue of public concern and the subject of active research. Africa with its high genetic diversity of honeybee sub-species and large wild population is also exposed to various factors responsible for colony losses in other parts of the world. Apart from the current American foulbrood epidemic in the Western Cape of South Africa, no large-scale colony losses have been reported elsewhere on the continent. We discuss the presence of pathogens, parasites, pests and predators of African honeybees as well as the threats they face in relation to habitat changes arising from the impact of increased human populations. In addition, we discuss current efforts aimed at protecting and promoting the health of African honeybees.

Infestation rates of Varroa destructor and Braula coeca in the savannah honey bee (Apis mellifera scutellata) Ursula Strauss, Christian W W Pirk, Vincent Dietemann, Robin M Crewe and Hannelie Human Social Insect Research Group, Department of Zoology and Entomology, University of Pretoria, Private Bag X20, Hatfield, Pretoria, South Africa 0028. Agroscope, Swiss Bee Research Centre, 3003 Bern, Switzerland.

Small hive beetles, Aethina tumida Murray, (Coleoptera: Nitidulidae) are native to subSaharan Africa where they parasitize honey bee (Apis mellifera L) colonies (Lundie 1940). Small hive beetles feed on pollen, brood and honey inside honey bee hives (Lundie 1940). Pupation occurs in the soil and the transitions from larvae to pupae to adult are vulnerable stages in the developmental cycle of small hive beetles (Lundie 1940; Hepburn

International audience; Pollen is the natural source of protein for bees and it is commonly assumed that a high protein content in pollen is beneficial. Investigation of the optimal nutrient ratio for honeybees was prompted by our earlier study showing surprisingly high mortality in caged honeybees fed with the protein-rich pollen of Aloe greatheadii var davyana, although field bees experience optimal growth when feeding on this pollen. We tested the effect of different protein sources and different protein:carbohydrate (P:C) ratios on the survival and ovarian activation of caged bees. Bees fed casein showed consistently higher survival than those fed royal jelly or aloe pollen, regardless of P:C ratios. They survived longer on lower P:C ratios and longest on a pure carbohydrate diet. The greatest ovarian activation was recorded for bees fed royal jelly in a 1:3 P:C ratio, showing the superior quality of royal jelly for supporting development.

Aloe marlothii and A. greatheadii var. davyana are two sympatric winter-flowering succulents that occur in the summer rainfall regions of northern and north-eastern South Africa. Both have flower characteristics that are strongly suggestive of bird pollination, although their nectar differs in volume and concentration. We conducted pollinator exclusion experiments to determine the importance of birds and insects as pollinators of these Aloe species. For both species fruit set and the number of seeds per fruit were higher in control treatments (all pollinators) and lower in treatments that excluded all pollinators. The contribution of insect pollinators to fruit set in A. marlothii was low (3–4%), like that of no pollinators (0–2%) whilst that of all pollinators (14–19%) was significantly higher, suggesting that generalist avian pollinators, which visited flowers in large numbers, are the most important pollinators. In A. greatheadii var. davyana fruit set in the absence of pollinators was also very low (2–6%), while the contribution to fruit set by insects (36–51%) was similar to that of all pollinators (55–55%), confirming the importance of honeybees to pollination. Clear understanding of both flower and nectar characteristics, and observations of flower visitors, are therefore required before an accurate prediction of pollinator type can be made. © 2009 SAAB. Published by Elsevier B.V. All rights reserved.

The winter‐flowering Aloe greatheadii var. davyana is a major indigenous bee plant in South Africa, widely distributed across the northern summer rainfall areas. Migratory beekeepers take advantage of its highly nutritious pollen for colony increase and strong nectar flow for honey production. We looked at variation on different levels in assessing this nectar resource for bees. There were no significant differences in nectar volume and concentration between the basal swelling (bulb) and the floral tube, only between flower stages. Nectar was continuously available, with volume and concentration remaining relatively constant throughout the day despite pronounced diurnal temperature changes and very low afternoon humidities. Bee foraging reduced mean nectar volumes in unscreened flowers by 50%, from 30.7 to 14.7 μL; bees are unable to access nectar in the bulb. Nectar volume was lowest and nectar concentration highest late in the flowering season, while the highest sugar content (3.54 mg per flower) was re...

Protein-rich diets are known to promote ovarian and egg development in workers of the honeybee, Apis mellifera, even in the presence of a queen. Since the main source of protein for honeybees is pollen, its quality and digestibility might be important dietary factors determining reproductive capacity. We have compared the effect of two types of pollen-sunflower, Helianthus annuus, and aloe, Aloe greatheadii var davyana-on ovarian development in A. mellifera scutellata workers. Under queenright conditions in the field, worker bees exhibited greater ovarian development when feeding on aloe pollen than on sunflower pollen. In their midgut, we observed higher extraction efficiency for aloe (80%) than for sunflower (69%) pollen. This may be attributed to the morphology and size of the two kinds of pollen grains and explains, together with the high protein content of aloe pollen (32% dry mass in bee-collected pollen) compared to sunflower pollen (15%), why aloe pollen promoted higher ovarian development. However, in the laboratory workers sustained on aloe pollen had significantly less-developed ovaries and higher mortality than those fed sunflower pollen. These detrimental effects may be due to an unbalanced protein:carbohydrate ratio. We discuss the effects of unbalanced diets on the physiology and ecology of honeybee reproduction.

This paper deals with the nectary structure and nectar presentation of two species belonging to different sections of the genus Aloe: A. castanea (Anguialoe) and A. greatheadii var. davyana (Pictae). The development of the nectary was studied by means of bright field and fluorescence light microscopy and scanning electron microscopy (SEM) in three flower stages (young, intermediate, old). Both species have septal nectaries. In A. castanea, a subsidiary tissue, not present in A. greatheadii var. davyana, was found beneath the nectary epithelium. This tissue accumulated starch that was hydrolyzed during secretion. Starch was slightly accumulated around the nectary in A. greatheadii var. davyana. The distribution of chlorophyll in the ovary was also different in the two species. These anatomical differences are not, however, correlated with greater nectar production in A. castanea. In this species, the nectary seems to degenerate after secretion, while in A. greatheadii var. davyana no sign of degeneration was observed. Differences in nectar presentation among the two species may account for different pollinators visiting their flowers.

We investigated the mechanism and efficiency of digestion of two types of pollen, maize, Zea mays, and sunflower, Helianthus annuus, by the spotted maize beetle, Astylus atromaculatus (Melyridae). We found similar and high extraction efficiencies, but different mechanisms of digestion. Osmotic shock was apparently involved in digestion of the large and thin-walled maize pollen grains. In the anterior midgut most maize pollen grains were already ruptured, in contrast with the intact exines of sunflower pollen, which suggests another mechanism of digestion for the latter, such as enzymatic action. We investigated the effect of osmotic shock on maize pollen in vitro by looking at the behavior of pollen grains in varying osmotic concentrations. Maize pollen grains burst in both distilled water and sugar solutions of various concentrations, and the amount of rupturing decreased with an increase in sugar concentration. Digestion of maize pollen was much slower in honeybees than in spotted maize beetles. Maize pollen bursts early in the midgut of maize beetles, but remains intact in honeybees: this suggests that osmotic shock may not be as important for honeybees as previously suggested.

Nectar concentration is assumed to remain constant during transport by honeybees between flowers and hive. We sampled crop contents of nectar foragers onAloe greatheadiivar.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.

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.

A variety of methods are used in honey bee research and differ depending on the level at which the research is conducted. On an individual level, the handling of individual honey bees, including the queen, larvae and pupae are required. There are different methods for the immobilising, killing and storing as well as determining individual weight of bees. The precise timing of developmental stages is also an important aspect of sampling individuals for experiments. In order to investigate and manipulate functional processes in honey bees, e. g. memory formation and retrieval and gene expression, microinjection is often used. A method that is used by both researchers and beekeepers is the marking of queens that serves not only to help to locate her during her life, but also enables the dating of queens. Creating multiple queen colonies allows the beekeeper to maintain spare queens, increase brood production or ask questions related to reproduction. On colony level, very useful techniques are the measurement of intra hive mortality using dead bee traps, weighing of full hives, collecting pollen and nectar, and digital monitoring of brood development via location recognition. At the population level, estimation of population density is essential to evaluate the health status and using beelines help to locate wild colonies. These methods, described in this paper, are especially valuable when investigating the effects of pesticide applications, environmental pollution and diseases on colony survival.

International audience; The nutritional needs of bees are receiving renewed attention in the context of declining bee populations and changes in land use that threaten floral resources. We present a comprehensive analysis of the nutritional composition of sunflower (Helianthus annuus L.) pollen, comparing hand-collected, bee-collected and stored pollen. As found in previous studies, the protein content of sunflower pollen was relatively low compared to other important bee forage plants. In the cultivars tested, two essential amino acids, methionine and tryptophan, are likely to be below the minimum requirements for honeybees. Fatty acid composition showed lauric acid to be most abundant, followed by palmitic and α-linolenic acids. While sunflower offers abundant and accessible pollen, its quality may hinder bee development when it is an exclusive pollen source, and the cultivars of such mass-flowering crops may vary in value for pollinators.

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.

Sustaining apiculture worldwide has been threatened by bee diseases and unexplained colony losses. African honeybee populations seem healthier and no major losses have been reported despite the presence of all the major pests and diseases. The scattered colonies in the large wild population of the continent might ensure slow pathogen spread and thus protect the unmanaged colonies in comparison with the concentration of colonies in managed apiaries. Beekeeping and trade in bee products is responsible for spreading many diseases throughout the world. The recent outbreak of the bacterial disease American foulbrood (AFB) in South Africa is a matter of great concern. Despite a growing number of apiaries testing positive for AFB, no major colony losses have been reported yet. This could be based on higher disease resistance of African honeybees, but such a trait might not persist if pathogens accumulate in the hives. In the first part of this article we review what is known on the history, biology and epidemiology of AFB as well as the control methods available. We then argue that given the particular context of honeybees in Africa, protection policies need to be put in place to ensure that African honeybees remain healthy.