Robin M. Crewe

Fertility signalling and reproductive skew in queenless ants

Social insects often show an extreme reproductive skew. In queenless ants, colonies consist of morphologically identical workers that can all potentially reproduce sexually. Similarly to that in social vertebrates, aggression in these ants functions to select the reproductive(s). We investigated the mechanisms underlying reproductive skew in the monogynous queenless ant Streblognathus peetersi. Behavioural observations of disturbed hierarchies were integrated with physiological measures of fertility (vitellogenin titre in the haemolymph) and chemical analysis of cuticular hydrocarbons, which are putative fertility pheromones. This multifaceted approach revealed that the colony reproductive is determined as a result of aggression between high-ranking workers, but once an alpha is established, chemical signalling is enough to maintain reproductive skew. As already reported in several species of ants and also in a social wasp, egg layers have distinct profiles of cuticular hydrocarbons compared with infertile workers. Importantly, ‘high rankers’ who are unable to lay eggs also have a specific cuticular profile; this is consistent with their intermediate state of fertility indicated by vitellogenin levels. Rather than just ovarian activity, the cuticular hydrocarbon profile thus reveals the individual hormonal state that underlies reproductive activity. We compare the fertility signal in queenless ants with the ‘badge of status’ reported in various birds. We discuss the evolutionary stability of this communication system and give special emphasis to ecological constraints and the high degree of intracolonial relatedness typical of social insects.

Estimating the Density of Honeybee Colonies across Their Natural Range to Fill the Gap in Pollinator Decline Censuses

Although pollinator declines are a global biodiversity threat, the demography of the western honeybee (Apis mellifera) has not been considered by conservationists because it is biased by the activity of beekeepers. To fill this gap in pollinator decline censuses and to provide a broad picture of the current status of honeybees across their natural range, we used microsatellite genetic markers to estimate colony densities and genetic diversity at different locations in Europe, Africa, and central Asia that had different patterns of land use. Genetic diversity and colony densities were highest in South Africa and lowest in Northern Europe and were correlated with mean annual temperature. Confounding factors not related to climate, however, are also likely to influence genetic diversity and colony densities in honeybee populations. Land use showed a significantly negative influence over genetic diversity and the density of honeybee colonies over all sampling locations. In Europe honeybees sampled in nature reserves had genetic diversity and colony densities similar to those sampled in agricultural landscapes, which suggests that the former are not wild but may have come from managed hives. Other results also support this idea: putative wild bees were rare in our European samples, and the mean estimated density of honeybee colonies on the continent closely resembled the reported mean number of managed hives. Current densities of European honeybee populations are in the same range as those found in the adverse climatic conditions of the Kalahari and Saharan deserts, which suggests that beekeeping activities do not compensate for the loss of wild colonies. Our findings highlight the importance of reconsidering the conservation status of honeybees in Europe and of regarding beekeeping not only as a profitable business for producing honey, but also as an essential component of biodiversity conservation.

The size of wild honeybee populations ( Apis mellifera ) and its implications for the conservation of honeybees

The density of wild honeybee colonies (Apis mellifera) in the African dry highland savannahs was estimated in three Nature Reserves in Gauteng, South Africa (Ezemvelo, Leeuwfontein, Suikerbosrand) based on the genotypes of drones which were caught at drone congregation areas. Densities were estimated to range between 12.4 and 17.6 colonies per square kilometer. In addition colony densities were estimated in two German National parks (Müritz and Hochharz) and a commercial mating apiary. The density of colonies was significantly lower at the German sampling sites with estimates of 2.4–3.2 colonies per square kilometer, which closely matches the nation-wide density of colonies kept by beekeepers. This shows that the densities of colonies observed in wild populations under the harsh conditions of the African dry savannahs exceeds that of Germany by far, in spite of intensive beekeeping. The intensity of apiculture in Europe is therefore unlikely to compensate for the loss of habitats suitable for wild honeybees due to agriculture, forestry and other cultivation of land.

Control of reproductive dominance by the thelytoky gene in honeybees

Differentiation into castes and reproductive division of labour are a characteristics of eusocial insects. Caste determination occurs at an early stage of larval development in social bees and is achieved via differential nutrition irrespective of the genotype. Workers are usually subordinate to the queen and altruistically refrain from reproduction. Workers of the Cape honeybee (Apis mellifera capensis) do not necessarily refrain from reproduction. They have the unique ability to produce female offspring parthenogenetically (thelytoky) and can develop into ‘pseudoqueens’. Although these are morphologically workers, they develop a queen-like phenotype with respect to physiology and behaviour. Thelytoky is determined by a single gene (th) and we show that this gene also influences other traits related to the queen phenotype, including egg production and queen pheromone synthesis. Using 566 microsatellite markers, we mapped this gene to chromosome 13 and identified a candidate locus thelytoky, similar to grainy head (a transcription factor), which has been shown to be highly expressed in queens of eusocial insects. We therefore suggest that this gene is not only important for determining the pseudoqueen phenotype in A. m. capensis workers, but is also of general importance in regulating the gene cascades controlling reproduction and sterility in female social bees.

Is there a need for conservation of honeybees in Africa

Honeybees are native to Africa and Europe but have been spread worldwide as the basis for an apicultural industry. To date, large and diverse wild populations only remain in Africa. On this continent the beekeeping industry is relatively undeveloped and relies on trapping swarms from wild populations to constitute the managed stocks. Bee breeding is seldom practiced. The situation is therefore different from that of Europe or North America where wild or feral honeybees have almost disappeared and this distinction is important when assessing the conservation status of African honeybees. While African honeybees appear to be more resistant to major diseases, the history of honeybee populations worldwide suggests that their conservation is a necessity. After analyzing the threats to which honeybees are exposed in Africa, we argue that preventive conservation measures are required to maintain the present favorable situation and avoid the declines in populations experienced elsewhere.ZusammenfassungDie Beziehung zwischen Menschen und Honigbienen hat in Afrika verschiedene Formen: In vielen Ländern wird Honigjagd praktiziert; In einigen Ländern ist die traditionelle Bienenhaltung in Körben oder Klotzbeuten weitverbreitet während andere moderne Bienenhaltungstechnologie einsetzen. Der hauptsächliche kontextuelle Unterschied zu anderen Weltregionen, in denen die Honigbienen entweder heimisch sind oder eingeführt wurden, ist die bedeutende Population von freilebenden Bienen, auf denen Honigjagd und Ausbeutung basieren. Im Gegensatz zu Europa machen bewirtschaftete Völker nur einen kleinen Teil der Honigbienenpopulation aus (14–18 aus 310 Mio). Weiterhin basiert die Bienenhaltung in Afrika weitgehend auf dem Fang von wilden Schwärmen und es wird keine oder nur wenig Bienenzucht betrieben. Diese Unterschiede sind wichtig, wenn die Honigbienenpopulationen von den in anderen Teilen der Welt zu beobachtenden Abnahmen geschützt werden sollen. Für Afrika sollten daher besondere politische Maßnahmen entwickelt werden. Auf diesem Kontinent sind die Netzwerke, auf die sich Bienenhalter und die Bienenindustrie stützen können nur sehr schwach entwickelt, und es wurden nur wenige Untersuchungen über die Honigbienen durchgeführt. Es ist daher sehr schwer, genaue Informationen und Zahlen über den Gesundheitszustand der Honigbienenpopulationen zu erhalten und daraus abzuleiten, ob Schutzmaßnahmen notwendig sind. Angesichts der Größe der Wildpopulation und dem Fehlen von Berichten über Epidemien könnte man schließen, dass die Wildpopulation in Afrika nicht gefährdet ist und daher von den Imkern ausgebeutet werden kann. Obwohl afrikanische Honigbienen den meisten Krankheiten ausgesetzt sind, die weltweit Honigbienen schädigen und sie diesen widerstehen können, sind der Verlust von Habitaten und Bejagung zwei Faktoren, denen europäische Bienen in geringerem Ausmaß ausgesetzt sind (da freilebende Honigbienen praktisch nicht mehr vorkommen und die bewirtschafteten Völker in vom Menschen hergestellten Beuten leben). Diese und andere noch nicht identifizierte Faktoren (z. B. neue Krankheiten) können Honigbienen negativ beeinflussen und ihre Wirkung könnte unter Umständen zu Verlusten in ähnlichem Ausmaß führen, wie sie andernorts beobachtet werden. Um zu verhindern, dass etwas Ähnliches in Afrika passieren kann, schlagen wir die Schaffung und Verbesserung sowie eine effektive Inkraftsetzung von Regulationen vor, die auf eine Erhaltung der afrikanischen Honigbienen abzielen. Da Afrika das Ursprungsland von Apis mellifera darstellt und das einzige Weltgebiet, in dem eine große Wildpopulation erhalten ist, sind Anstrengungen zur Erhaltung nicht nur zum Erhalt von einzigartigen Populationen vonnöten, sondern auch, um das Funktionieren des Ökosystems und der landwirtschaftlichen Produktion sicherzustellen, die beide von den Bestäubungsleistungen der Honigbienen abhängig sind.

A survey of managed honey bee colony losses in the Republic of South Africa–2009 to 2011

Summary This 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 causes of mortality were small hive beetles, varroa mites, absconding (non-reproductive swarming), and chalkbrood disease. Those listing chalkbrood disease lost significantly fewer colonies than those who did not list chalkbrood. The exact mechanism for this difference is unknown, and may be related to other beekeeping practices that correlate with finding chalkbrood infections—namely more intensive inspection and management.

Alternative splicing of a single transcription factor drives selfish reproductive behavior in honeybee workers (Apis mellifera)

In eusocial insects the production of daughters is generally restricted to mated queens, and unmated workers are functionally sterile. The evolution of this worker sterility has been plausibly explained by kin selection theory [Hamilton W (1964) J Theor Biol 7:1–52], and many traits have evolved to prevent conflict over reproduction among the females in an insect colony. In honeybees (Apis mellifera), worker reproduction is regulated by the queen, brood pheromones, and worker policing. However, workers of the Cape honeybee, Apis mellifera capensis, can evade this control and establish themselves as social parasites by activating their ovaries, parthenogenetically producing diploid female offspring (thelytoky) and producing queen-like amounts of queen pheromones. All these traits have been shown to be strongly influenced by a single locus on chromosome 13 [Lattorff HMG, et al. (2007) Biol Lett 3:292–295]. We screened this region for candidate genes and found that alternative splicing of a gene homologous to the gemini transcription factor of Drosophila controls worker sterility. Knocking out the critical exon in a series of RNAi experiments resulted in rapid worker ovary activation—one of the traits characteristic of the social parasites. This genetic switch may be controlled by a short intronic splice enhancer motif of nine nucleotides attached to the alternative splice site. The lack of this motif in parasitic Cape honeybee clones suggests that the removal of nine nucleotides from the altruistic worker genome may be sufficient to turn a honeybee from an altruistic worker into a parasite.

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

SUMMARY 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.

Pheromonal contest between honeybee workers (Apis mellifera capensis)

Abstract Queenless workers of the Cape honeybee (Apis mellifera capensis) can develop into reproductives termed pseudoqueens. Although they morphologically remain workers they become physiologically queenlike, produce offspring, and secrete mandibular gland pheromones similar to those of true queens. However, after queen loss only very few workers gain pseudoqueen status. A strong intracolonial selection governs which workers start oviposition and which remain sterile. The “queen substance”, 9-keto-2(E)-decenoic acid (9-ODA), the dominant compound of the queens mandibular gland pheromones, suppresses the secretion of queenlike mandibular gland pheromones in workers. It may act as an important signal in pseudoqueen selection. By analysing the mandibular gland pheromones of workers kept in pairs, we found that A. m. capensis workers compete to produce the strongest queen-like signal.

The role of the queen mandibular gland pheromone in honeybees (Apis mellifera): honest signal or suppressive agent?

Queen pheromones interfere with worker reproduction in social insects. However, there is still an unresolved question as to whether this pheromone acts as an “honest” signal for workers, giving a reliable indication of the queen’s reproductive value, or as a suppressive agent, inhibiting worker reproduction independent of the queen’s reproductive capacity. In honeybees (Apis mellifera), the queen’s mandibular gland secretion, a mix of fatty acids and some aromatic compounds, is crucial for regulating the reproductive division of labor in the colony inhibiting ovary development in workers. We quantified the mandibular gland secretions of virgin, drone-laying, and naturally mated queens using gas chromatography to test whether the queens’ mating, ovary activation, or the reproductive value for workers correlated with the composition of the secretion. Although the absolute amounts of the “queen substance” 9-oxo-2(E)-decenoic acid (9-ODA) were similar among the three groups, the proportions of 9-ODA decreased with increasing reproductive quality. Furthermore, the ratios of queen to worker compounds were similar in all three treatment groups, irrespective of the reproductive capacity. A multivariate analysis including all six compounds could not separate drone-laying queens from naturally mated ones, both with active ovaries but only the latter ensuring colony survival. We suggest that the mandibular gland pheromones are unlikely to function as reliable indicators of queen reproductive value and rather operate as an agent to suppress worker reproduction. This does not exclude the possibility that other “honest” pheromone signals exist in the honeybee colony, but these would have to arise from other semiochemicals, which could be produced by both the queen and the brood.