Reproduction of Varroa destructor in worker brood of Africanized honey bees (Apis mellifera)

Reproduction and population growth of Varroa destructor was studied in ten naturally infested, Africanized honeybee (AHB) (Apis mellifera) colonies in Yucatan, Mexico. Between February 1997 and January 1998 monthly records of the amount of pollen, honey, sealed worker and drone brood were recorded. In addition, mite infestation levels of adult bees and worker brood and the fecundity of the mites reproducing in worker cells were determined. The mean number of sealed worker brood cells (10,070 ± 1,790) remained fairly constant over the experimental period in each colony. However, the presence and amount of sealed drone brood was very variable. One colony had drone brood for 10 months and another for only 1 month. Both the mean infestation level of worker brood (18.1 ± 8.4%) and adult bees (3.5 ± 1.3%) remained fairly constant over the study period and did not increase rapidly as is normally observed in European honey bees. In fact, the estimated mean number of mites fell from 3,500 in February 1997 to 2,380 in January 1998. In May 2000 the mean mite population in the study colonies was still only 1,821 mites. The fertility level of mites in this study was much higher (83–96%) than in AHB in Brazil(25–57%), and similar to that found in EHB (76–94%). Mite fertility remained high throughout the entire study and was not influenced by the amount of pollen, honey or worker brood in the colonies. This revised version was published online in July 2006 with corrections to the Cover Date.     

Self-assemblage formation in a social insect: the protective curtain of a honey bee swarm

Summary. This paper considers a little-studied topic in the biology of social insects: the formation of self-assemblages. It focuses on the mechanisms whereby the outermost workers in a bivouacked swarm of honey bees, when rained upon, form a water repellent curtain of bees over the swarm cluster. Specifically, we analyzed how the worker bees in the mantle of a swarm cluster adjust their body orientation, wing spread, and inter-individual spacing to form a protective curtain when wetted. When warm and dry, the mantle bees orient their bodies weakly with respect to gravity, do not tuck their heads under adjacent bees, have high variability in wing spread, and space themselves widely. In contrast, when warm and wet, the mantle bees orient uniformly with head upward, tuck their heads beneath the abdomens of bees above, hold their wings together, and press tightly together. This produces a surface that closely resembles a tiled roof. When cool and dry, the mantle bees generally orient their bodies with head upward, press their heads into the interior of the cluster, hold their wings wide apart, and draw close together. We also examined the age distribution of the mantle bees. Older bees are more likely than younger bees to be found in the mantle of a swarm, perhaps because younger bees are more important than older bees to colony survival after swarming and so occupy a more sheltered position in a swarm. Finally, we tested whether swarm clusters that have formed a protective curtain shed water more effectively than ones that have not formed a curtain. We found that this is the case.Received 28 November 2003; revised 29 February 2004; accepted 11 April 2004.     

Host-seeking behaviour of tracheal mites (Acari: Tarsonemidae) on honey bees (Hymenoptera: Apidae)

The behaviour of the endoparasitic tracheal mite, Acarapis woodi (Rennie) on honey bees (Apis mellifera L.) is a challenge to observe because of its small size. Through a microscope, we videotaped this mite's movement on young bees, dead bees and bees exposed to vegetable oil. Previous studies have shown that solid vegetable oil decreases mite infestations in a bee colony. We hypothesized that the oil alters mite behaviour to the detriment of the parasite, thus helping to safeguard the host. Habitat-seeking behaviour, identified as necessary for mites to locate a new host environment, was disrupted on both dead and oil-treated bees. Questing behaviour, which is associated with transfer between hosts, increased significantly on the dead and oily bees. The behaviours of mites were significantly different between all three treatments (x ²=494.96, p<0.001 on dead bees and x ²=851.11, p<0.001 on oily bees). Both questing and seeking behaviours were significantly different on each of the thoracic treatments (F _2,66=7.88, p<0.001 and F _2,66=21.28, p<0.001) and mite questing behaviour was not altered between males and females on live or oily bees (F _1,22=0.25, p<0.62), but habitat seeking was (F _1,22=7.42, p<0.012). The male questing and habitat-seeking behaviours were observed. We conclude that oil-treated bees gained protection from habitat-seeking mites because the normal behaviour of the mites seeking an oviposition site is interrupted.     

Alarm pheromone perception in honey bees is decreased by smoke (Hymenoptera: Apidae)

The application of smoke to honey bee(Apis mellifera) antennae reduced the subsequent electroantennograph response of the antennae to honey bee alarm pheromones, isopentyl acetate, and 2-heptanone. This effect was reversible, and the responsiveness of antennae gradually returned to that of controls within 10–20 min. A similar effect occurred with a floral odor, phenylacetaldehyde, suggesting that smoke interferes with olfaction generally, rather than specifically with honey bee alarm pheromones. A reduction in peripheral sensitivity appears to be one component of the mechanism by which smoke reduces nest defense behavior of honey bees.     

Nonrandom visitation of brood cells by worker honey bees (Hymenoptera: Apidae)

The visitation pattern by worker honey bees to cells in the brood nest was monitored on an artificially created brood pattern consisting of about one-fourth brood cells evenly distributed among empty cells. The majority (63 %) of the observed workers selectively entered larval cells. In contrast, some workers avoided egg cells when presented a choice of egg vs empty cells. The results suggest that larvae produce a general signal indicating their presence to worker bees. Eggs also seem to produce a signal, which is perceived to be different from the one from larvae.     

Nosema ceranae has been present in Brazil for more than three decades infecting Africanized honey bees

Until the mid-1990s, the only microsporidium known to infect bees of the genus Apis was Nosema apis. A second species, Nosema ceranae, was first identified in 1996 from Asian honey bees; it is postulated that this parasite was transmitted from the Asian honey bee, Apis cerana, to the European honey bee, Apis mellifera. Currently, N. ceranae is found on all continents and has often been associated with honey bee colony collapse and other reports of high bee losses. Samples of Africanized drones collected in 1979, preserved in alcohol, were analyzed by light microscopy to count spores and were subjected to DNA extraction, after which duplex PCR was conducted. All molecular analyses (triplicate) indicated that the drones were infected with both N. ceranae and N. apis. PCR products were sequenced and matched to sequences reported in the GenBank (Acc. Nos. JQ639316.1 and JQ639301.1). The venation pattern of the wings of these males was compared to those of the current population living in the same area and with the pattern of drones collected in 1968 from Ribeirão Preto, SP, Brazil, from a location close to where African swarms first escaped in 1956. The morphometric results indicated that the population collected in 1979 was significantly different from the current living population, confirming its antiquity. Considering that the use of molecular tools for identifying Nosema species is relatively recent, it is possible that previous reports of infections (which used only light microscopy, without ultrastructural analysis) wrongly identified N. ceranae as N. apis. Although we can conclude that N. ceranae has been affecting Africanized honeybees in Brazil for at least 34 years, the impact of this pathogen remains unclear.     

Admixture in Africanized honey bees (Apis mellifera) from Panamá to San Diego,California (U.S.A.)

The Africanized honey bee (AHB) is a New World amalgamation of several subspecies of the western honey bee (Apis mellifera), a diverse taxon historically grouped into four major biogeographic lineages: A (African), M (Western European), C (Eastern European), and O (Middle Eastern). In 1956, accidental release of experimentally bred “Africanized” hybrids from a research apiary in Sao Paulo, Brazil initiated a hybrid species expansion that now extends from northern Argentina to northern California (U.S.A.). Here, we assess nuclear admixture and mitochondrial ancestry in 60 bees from four countries (Panamá; Costa Rica, Mexico; U.S.A) across this expansive range to assess ancestry of AHB several decades following initial introduction and test the prediction that African ancestry decreases with increasing latitude. We find that AHB nuclear genomes from Central America and Mexico have predominately African genomes (76%–89%) with smaller contributions from Western and Eastern European lineages. Similarly, nearly all honey bees from Central America and Mexico possess mitochondrial ancestry from the African lineage with few individuals having European mitochondria. In contrast, AHB from San Diego (CA) shows markedly lower African ancestry (38%) with substantial genomic contributions from all four major honey bee lineages and mitochondrial ancestry from all four clades as well. Genetic diversity measures from all New World populations equal or exceed those of ancestral populations. Interestingly, the feral honey bee population of San Diego emerges as a reservoir of diverse admixture and high genetic diversity, making it a potentially rich source of genetic material for honey bee breeding.     

Do honey bees encode distance information into the wing vibrations of the waggle dance?

An optical technique detected the wing vibration frequency of worker honey bees in an observation hive during the straight run of the waggle dance. Wing oscillation frequencies were recorded from dancing bees after they had visited a feeding station located from 50 to 1600 m from the hive. The bees vibrated their wings more rapidly after they visited nearby stations than when they foraged at more distant feeding stations. For example, the mean frequency of 315 Hz at 50 m dropped to only 207 Hz at 1600 m. Wing vibration frequency appears to be another factor to be added to the elements in the dance known to indicate the distance bees must fly to food sources. These known elements include the duration of the straight run and the number of wagtail movements in the run.     

Selection on a haploid genotype for discrimination learning performance: Correlation between drone honey bees (Apis mellifera) and their worker progeny (Hymenoptera: Apidae)

Successful bidirectional selection for discriminative olfactory learning is reported for drone honey bees (Apis mellifera). Learning performance was evaluated using a discrimination conditioning procedure that required drones to discriminate between an appetitively reinforced odorant and one that was followed by punishment. Selective breeding produced high- and low-learning-performance lines of worker progeny that diverged from performance of workers whose fathers were selected at random. Furthermore, we show that levels of sucrose-induced sensitization are not correlated to learning performance. These results corroborate earlier findings and further demonstrate the power of selection on a haploid (drone) genotype. In addition, this study now shows that the demonstrated differences in learning performance cannot be completely accounted for by alteration of sucrose-induced sensitization. Thus, using this technique, it may be possible to select for associative conditioning without a pleiotropic increase in sensitization. The honey bee will be ideally suited to these types of correlation analyses in future studies.     

Worker policing persists in a hopelessly queenless honey bee colony (<Emphasis Type="Italic">Apis mellifera</Emphasis>)

Summary In queenright colonies of Apis mellifera, worker policing normally eliminates worker-laid eggs thereby preventing worker reproduction. However, in queenless colonies that have failed to rear a replacement queen, worker reproduction is normal. Worker policing is switched off, many workers have active ovaries and lay eggs, and the colony rears a last batch of male brood before dying out. Here we report a colony which, when hopelessly queenless, did not stop policing although a high proportion of workers had active ovaries (12.6%) and many eggs were laid. However, all these eggs and also worker-laid eggs transferred from another colony were policed. This unusual pattern was repeated eight weeks later by a second queenless colony made using worker bees from the same mother colony, which strongly suggests genetic determination.Received 19 May 2003; revised 11 September 2003; accepted 23 September 2003.     

Muscarinic regulation of Kenyon cell dendritic arborizations in adult worker honey bees

The experience of foraging under natural conditions increases the volume of mushroom body neuropil in worker honey bees. A comparable increase in neuropil volume results from treatment of worker honey bees with pilocarpine, an agonist for muscarinic-type cholinergic receptors. A component of the neuropil growth induced by foraging experience is growth of dendrites in the collar region of the calyces. We show here, via analysis of Golgi-impregnated collar Kenyon cells with wedge arborizations, that significant increases in standard measures of dendritic complexity were also found in worker honey bees treated with pilocarpine. This result suggests that signaling via muscarinic-type receptors promotes the increase in Kenyon cell dendritic complexity associated with foraging. Treatment of worker honey bees with scopolamine, a muscarinic inhibitor, inhibited some aspects of dendritic growth. Spine density on the Kenyon cell dendrites varied with sampling location, with the distal portion of the dendritic field having greater total spine density than either the proximal or medial section. This observation may be functionally significant because of the stratified organization of projections from visual centers to the dendritic arborizations of the collar Kenyon cells. Pilocarpine treatment had no effect on the distribution of spines on dendrites of the collar Kenyon cells.     

Anarchistic queen honey bees have normal queen mandibular pheromones

Summary. Anarchistic honey bees are a line developed by recurrent selection in which workers frequently lay eggs. In unselected colonies, workers refrain from reproduction in response to pheromonal signals that indicate the presence of brood and a queen. We show that queen type (anarchistic or wild type) has no effect on rates of ovary activation of anarchistic or wild type workers. In addition, we show that an important component of the queens signalling system, the queen mandibular gland pheromone, is similar in wild type and anarchistic queens. Anarchistic larvae do not inhibit worker ovary development to the same degree as wild type larvae, however all colonies in this experiment contained only wild type larvae. Anarchistic workers had greater rates of ovary activation than wild type workers in colonies headed by either queen type. We therefore conclude that there must be differences in the transmission or reception of queen pheromones, or worker sensitivity to these compounds. These results clearly demonstrate that anarchy is a complex syndrome, not simply the result of reduced pheromone production by anarchist queens and larvae.Received 23 December 2003; revised 14 May 2004; accepted 1 June 2004.     

The waggle dance of the honey bee: Which bees following a dancer successfully acquire the information?

During the waggle dance of the honeybee, the dancer is able to tell her nestmates the distance and direction to a rich food source (Frisch, 1967). Little is known about how waggle dance followers are able to read the waggle dance in the darkness of a hive. Initial observations showed that not all of the bees that appear to be dance followers behave the same. Some bees maneuver themselves behind the dancer, while others do not. The paths of a single dancer, trained to an artificial food source, and her followers were traced during the waggle runs. The success of these dance followers was compared to their position relative to the dancer. The results of this study show that during a waggle run a dance follower must position itself within a 30° arc behind the dancer in order to obtain the dance information. The results suggest that bees are using the position of their own bodies to determine direction.     

The effects of season,pretraining, and scent on the efficiency of traps for capturing recruited honey bees (Hymenoptera: Apidae)

A portable trap was designed to capture honey bees recruited to the field by dancers. An infrared phototransistor placed in the entrance tunnel of the trap sensed an incoming bee and turned on a talking clock, which in turn activated a voice-actuated audio tape recorder that recorded the time. We tested the effectiveness of traps for capturing bees recruited by four dancer bees (1) during two seasons when local flower densities differed, (2) with or without a group of bees pretrained to enter traps for food, and (3) when the scent used in traps and at the dancers' feeding station was changed just prior to recruitment trials or was not changed. One trap was put out at each of four distances (50, 100, 150, and 200 m) from the hive, while dancers fed on concentrated sucrose at the feeding station located at 150 m in the same direction. Recruited bees that approached the traps but did not enter were counted by observers. More bees were recruited and captured in traps when the local flora was sparse (fall) than when flowers were abundant (summer), when bees were pretrained versus not pretrained, and when the scent was not changed just prior to recruitment trials versus changed. The distributions of number of bees counted at the four distances at scented recruit stations and trapped were similar only when bees were pretrained and the scent was not changed during recruitment trials. However, the highest proportion of bees trapped in a trial at 150 m (distance to dancers' feeding station) was when bees were pretrained and the scent was changed.     

Influence of flight time and flight environment on distance communication by dancing honey bees

Forager honey bees communicate the distance of food sources to nest mates through waggle dances, but how do bees measure these distances? Recent work suggests that bees measure distance flown in terms of the extent of image motion (integrated optic flow) that is experienced during flight. However, it is known that optic flow also regulates the speed of flight. Therefore, the duration of the flight to a destination is likely to co-vary with the optic flow that is experienced en route. This makes it difficult to tease apart the potential roles of flight duration and optic flow as cues in estimating distance flown. Here we examine whether flight duration alone can serve as an indicator of distance. We trained bees to visit feeders at two sites located in optically different environments, but positioned such that the flight durations to the two sites were similar. The distance estimates for the two sites, as reported in the waggle dance, were compared. We found that dances differed significantly between the two sites, even though flight times were similar. Flight time perse was a poor predictor of waggle dance behaviour. We conclude that foraging bees do not simply signal flight time as their measure of distance in the waggle dance; the environment through which they fly plays a dominant role. Received 11 April 2005; revised 16 May 2005; accepted 3 June 2005.     

Evidence for reproductive isolation between two colour morphs of cavity nesting honey bees (Apis) in south India

In south India there are two distinct colour morphs of cavity nesting honey bees: the yellow ‘Plain’ morph and the black ‘Hill’ morph which are collectively known as Apis cerana. We show that the Hill morph is associated with a widely distributed mitochondrial haplotype that is present throughout mainland populations of south east Asian A. cerana. In contrast, the Plain morph, which is apparently confined to low to moderate elevations in India and Sri Lanka, is associated with a unique mitochondrial haplotype that is not present in other cavity nesting honey bees. We further show that in a region of sympatry (Bangalore, Karnataka State) the drone mating flight times of the two colour morphs barely overlap. Combined, drone flight data and the complete separation of mitochondrial haplotypes suggest that the two morphs are reproductively isolated. The Plain morph is distinguished from the Hill morph by the first three abdominal tergites of the worker, which are completely yellow in the Plain morph, whereas in the Hill morph they are black or black with yellow patches. Although the two morphs are generally distinguishable in the field by overall colouration, microscopic examination of the first 3 abdominal tergites is preferred. Received 16 February 2006; revised 11 May 2006; accepted 23 May 2006.