Honey bee foraging: persistence to non-rewarding feeding locations and waggle dance communication

The honey bee, Apis mellifera, is important in agriculture and also as a model species in scientific research. This Master’s thesis is focused on honey bee foraging behaviour. It contains two independent experiments, each on a different subject within the area of foraging. Both use a behavioural ecology approach, with one investigating foraging behaviour and the other foraging communication. These form chapters 2 and 3 of the thesis, after an introductory chapter. Chapter 2. Experiment 1: Persistence to unrewarding feeding locations by forager honey bees (Apis mellifera): the effects of experience, resource profitability, and season This study shows that the persistence of honey bee foragers to unrewarding food sources, measured both in duration and number of visits, was greater to locations that previously offered sucrose solution of higher concentration (2 versus 1molar) or were closer to the hive (20 versus 450m). Persistence was also greater in bees which had longer access at the feeder before the syrup was terminated (2 versus 0.5h). These results indicate that persistence is greater for more rewarding locations. However, persistence was not higher in the season of lowest nectar availability in the environment. Chapter 3. Experiment 2: Honey bee waggle dance communication: signal meaning and signal noise affect dance follower behaviour This study shows that honey bee foragers follow fewer waggle runs as the distance to the food source, that is advertised by the dance, increases, but invest more time in following these dances. This is because waggle run duration increases with increasing foraging distance. The number of waggle runs followed for distant food sources was further reduced by increased angular noise among waggle runs within a dance. The number of dance followers per dancing bee was affected by the time of year and varied among colonies. Both noise in the message, that is variation in the direction component, and the message itself, that is the distance of the advertised food location, affect dance following. These results indicate that dance followers pay attention to the costs and benefits associated with using dance information

Integrated control of honey bee diseases in apiculture

The honey bee, Apis mellifera, is important both ecologically and economically. Pests and diseases are arguably the greatest current challenge faced by honey bees and beekeeping. This PhD thesis is focused on honey bee disease control including natural resistance by means of hygienic behaviour. It contains eleven independent experiments, ten on honey bee pests and diseases and their control and resistance, and one on stingless bees. Each is written as a separate chapter, Chapters 4 and 14 of this thesis. Chapter 4: How effective is Apistan® at killing varroa? This shows that Apistan is not very effective at killing varroa, presumably because of resistance. It also shows that a single Apistan treatment resulted in the next treatment being significantly less effective, indicating strong selection for resistance. Chapter 5: Towards integrated control of varroa: comparing application methods and doses of oxalic acid on the mortality of phoretic Varroa destructor mites and their honey bee hosts. This shows that oxalic acid can be highly effective at killing varroa mites under beekeeping conditions in broodless hives in winter. However, varroa mortality is affected by application method and dose. In addition, bee and colony mortality and colony performance are also affected by application method and dose. The results of this chapter shows that sublimation is the best method, in that it gives greater varroa mortality at lower doses, and results in no harm to the colonies. In fact, colonies treated via sublimation had significantly more brood in spring that controls, and lower winter mortality, although this difference was not significant. Chapter 6: Towards integrated control of varroa: varroa mortality from treating broodless winter colonies twice with oxalic acid via sublimation. This shows that two treatments of 2.25 g oxalic acid via sublimation at an interval of 2 weeks in broodless honey bee colonies in winter result in greater varroa mortality than a single treatment, 99.6% vs. 97%. Making a second oxalic acid treatment was not harmful as the performance (frames of brood, queen and colony survival) of the twice-treated colonies over the next 4 months was not significantly different to the once-treated control colonies. Chapter 7: Towards integrated control of varroa: Efficacy of early spring trapping in drone brood. This indicates that trapping in drone brood in spring is probably not sufficiently effective to be able to control varroa populations on its own. It shows that trapping varroa in capped drone cells in early spring is not highly effective at controlling varroa. The first and second test frames of drone foundation removed 44% and 48% of the varroa, respectively. Chapter 8: Towards integrated control of varroa: Monitoring honey bee brood rearing in winter and the proportion of varroa in small patches of sealed cells. This shows that December is the month with the least brood. However, winter reduction in brood rearing varied among years and even in December some colonies still had sealed brood. Although the amounts of sealed brood were low, even a small patch of c. 500-600 sealed cells could contain 14% of the varroa in a colony. This will halve the duration of control provided by an oxalic acid treatment. Chapter 9: Towards integrated control of varroa: effect of variation in hygienic behaviour among honey bee colonies on mite population increase and deformed wing virus incidence. This shows clearly that hygienic behaviour reduces the one-year population growth of varroa in honey bee colonies by more than 50% and reduces the levels of deformed wing virus by more than 1000 times. Chapter 10: Hygienic behaviour saves the lives of honey bee colonies. This shows that hygienic behaviour saves the lives of honey bee colonies with shrivelled wings, a visible symptom of deformed wing virus that is considered a predictor of colony death. Over one year, only 2 of 11 colonies requeened with a non-hygienic queen survived, versus 13 of 15 requeened with a hygienic queen. Chapter 11: Hygienic behaviour by non-hygienic honey bee colonies: all colonies remove dead brood from open cells. This shows that all honey bee colonies are highly hygienic in response to dead or diseased brood in open cells. All larvae killed by freezing with liquid nitrogen and larvae with chalkbrood disease were removed. This was true even for colonies with low levels of removal of dead brood from sealed cells, which would be considered as non-hygienic colonies. Chapter 12: Removal of larvae infected by different strains of chalk brood and other fungi by hygienic and non-hygienic bee colonies. This shows that hygienic and non-hygienic honey bee colonies are highly hygienic in response to diseased larvae killed with different strains of fungus in open cells. Chapter 13: Hygienic behaviour in Brazilian stingless bees. This shows that the three stingless bee species studied (Melipona scutellaris, Scaptotrigona depilis, Tetragonisca angustula) all have high levels of hygienic behaviour, quantified as the removal of freeze-killed brood, in comparison to the honey bee Apis mellifera. In S. depilis there was considerable variation in hygienic behaviour among colonies, and hygienic colonies removed more brood affected by a naturally-occurring disease which we discovered and for which the causative agent remains to be identified. Chapter 14: First record of small hive beetle, Aethina tumida Murray, in South America. This reports the discovery of adult small hive beetles, Aethina tumida, in honey bee, Apis mellifera, hives in an apiary in Brazil, in March 2015. This is the first record for South America of this honey bee pest

Towards integrated control of varroa: effect of variation in hygienic behaviour among honey bee colonies on mite population increase and deformed wing virus incidence

Hygienic behaviour in the honey bee, Apis mellifera, is the uncapping and removal of dead, diseased or infected brood from sealed cells by worker bees. We determined the effect of hygienic behaviour on varroa population growth and incidence of deformed wing virus (DWV), which can be transmitted by varroa. We treated 42 broodless honey bee colonies with oxalic acid in early January 2013 to reduce varroa populations to low levels, which we quantified by extracting mites from a sample of worker bees. We quantified varroa levels, again when the colonies were broodless, 48 weeks later. During the summer the hygienic behaviour in each colony was quantified four times using the Freeze Killed Brood (FKB) removal assay, and ranged from 27.5 % to 100 %. Varroa population increased greatly over the season, and there was a significant negative correlation between varroa increase and FKB removal. This was entirely due to fully hygienic colonies with >95 % FKB having only 43 % of the varroa build up of the less hygienic colonies.None of the 14 colonies with >80 % FKB removal had overt symptoms of DWV, whilst 36 % of the less hygienic colonies did. Higher levels of FKB removal also correlated significantly with lower numbers of DWV RNA copies in worker bees, but not in varroa mites. On average, fully hygienic colonies had c. 10,000 times less viral RNA than less hygienic colonies

Appetite for self-destruction: suicidal biting as a nest defense strategy in Trigona stingless bees

Self-sacrificial behavior represents an extreme and relatively uncommon form of altruism in worker insects. It can occur, however, when inclusive fitness benefits are high, such as when defending the nest. We studied nest defense behaviors in stingless bees, which live in eusocial colonies subject to predation. We introduced a target flag to nest entrances to elicit defensive responses and quantified four measures of defensivity in 12 stingless bee species in São Paulo State, Brazil. These included three Trigona species, which are locally known for their aggression. Species varied significantly in their attack probability (cross species range = 0–1, P < 0.001), attack latency (7.0–23.5 s, P = 0.002), biting duration of individual bees (3.5–508.7 s, P < 0.001), and number of attackers (1.0–10.8, P < 0.001). A “suicide” bioassay on the six most aggressive species determined the proportion of workers willing to suffer fatal damage rather than disengage from an intruder. All six species had at least some suicidal individuals (7–83 %, P < 0.001), reaching 83 % in Trigona hyalinata. Biting pain was positively correlated with an index of overall aggression (P = 0.002). Microscopic examination revealed that all three Trigona species had five sharp teeth per mandible, a possible defensive adaptation and cause of increased pain. Suicidal defense via biting is a new example of self-sacrificial altruism and has both parallels and differences with other self-sacrificial worker insects, such as the honey bee. Our results indicate that suicidal biting may be a widespread defense strategy in stingless bees, but it is not universal

Quality versus quantity: foraging decisions in the honeybee (Apis mellifera scutellata) feeding on wildflower nectar and fruit juice

Foraging animals must often decide among resources which vary in quality and quantity. Nectar is a resource that exists along a continuum of quality in terms of sugar concentration and is the primary energy source for bees. Alternative sugar sources exist, including fruit juice, which generally has lower energetic value than nectar. We observed many honeybees (Apis mellifera scutellata) foraging on juice from fallen guava (Psidium guajava) fruit near others foraging on nectar. To investigate whether fruit and nectar offered contrasting benefits of quality and quantity, we compared honeybee foraging performance on P. guajava fruit versus two wildflowers growing within 50 m, Richardia brasiliensis and Tridax procumbens. Bees gained weight significantly faster on fruit, 2.72 mg/min, than on either flower (0.17 and 0.12 mg/min, respectively). However, the crop sugar concentration of fruit foragers was significantly lower than for either flower (12.4% vs. 37.0% and 22.7%, respectively). Fruit foragers also spent the most time handling and the least time flying, suggesting that fruit juice was energetically inexpensive to collect. We interpret honeybee foraging decisions in the context of existing foraging models and consider how nest-patch distance may be a key factor for central place foragers choosing between resources of contrasting quality and quantity. We also discuss how dilute solutions, such as fruit juice, can help maintain colony sugar–water balance. These results show the benefits of feeding on resources with contrasting quality and quantity and that even low-quality resources have value

Honey bee waggle dances facilitate shorter foraging distances and increased foraging aggregation

Many social organisms assist their group mates in foraging. The honey bee waggle dance allows nestmates to communicate the locations of rewarding flower patches. This remarkable behaviour can increase colony food collection under certain environmental conditions. However, we know little about how it affects colony foraging distribution in the landscape. Here, we predicted that dance communication influences foraging distances and increases the aggregation of foraging locations. We tested these predictions by manipulating the ability of Apis mellifera colonies situated in a heterogeneous landscape to communicate location information. Following established methodology, six study colonies in observation hives were turned horizontally and exposed to light treatments that produced either oriented or disoriented dancing. After 10–17 days, hives were then turned vertically and allowed to dance under normal conditions. We videoed and decoded dances to compare foraging patterns that had developed during the preceding treatment. This procedure was repeated throughout the foraging season from spring to autumn. Dance decoding revealed that, on average, bees from disoriented colonies recruited for food sources further away than oriented colonies. This effect was consistent throughout the year. Oriented colonies' recruitment was to a smaller area and was more spatially aggregated than that of disoriented colonies. However, experimental treatment did not affect colony weight gain. Our results show that dance communication plays an important role in the spatial distribution of foraging and is potentially beneficial in reducing commuting costs by directing recruits to closer foraging locations. These findings may be particularly relevant in heterogeneous landscapes.</p

Hygienic behaviour in Brazilian stingless bees

Social insects have many defence mechanisms against pests and pathogens. One of these is hygienic behaviour, which has been studied in detail in the honey bee, Apis mellifera. Hygienic honey bee workers remove dead and diseased larvae and pupae from sealed brood cells, thereby reducing disease transfer within the colony. Stingless bees, Meliponini, also rear broods in sealed cells. We investigated hygienic behaviour in three species of Brazilian stingless bees (Melipona scutellaris, Scaptotrigona depilis, Tetragonisca angustula) in response to freeze-killed brood. All three species had high mean levels of freeze-killed brood removal after 48 h ∼99% in M. scutellaris, 80% in S. depilis and 62% in T. angustula (N=8 colonies per species; three trials per colony). These levels are greater than in unselected honey bee populations, ∼46%. In S. depilis there was also considerable intercolony variation, ranging from 27% to 100% removal after 2 days. Interestingly, in the S. depilis colony with the slowest removal of freeze-killed brood, 15% of the adult bees emerging from their cells had shrivelled wings indicating a disease or disorder, which is as yet unidentified. Although the gross symptoms resembled the effects of deformed wing virus in the honey bee, this virus was not detected in the samples. When brood comb from the diseased colony was introduced to the other S. depilis colonies, there was a significant negative correlation between freeze-killed brood removal and the emergence of deformed worker bees (P=0.001), and a positive correlation with the cleaning out of brood cells (P=0.0008). This shows that the more hygienic colonies were detecting and removing unhealthy brood prior to adult emergence. Our results indicate that hygienic behaviour may play an important role in colony health in stingless bees. The low levels of disease normally seen in stingless bees may be because they have effective mechanisms of disease management, not because they lack diseases

Landscape scale study of the net effect of proximity to a neonicotinoid-treated crop on bee Colony health

Since 2013, the European Commission has restricted the use of three neonicotinoid insecticides as seed dressings on bee-attractive crops. Such crops represent an important source of forage for bees, which is often scarce in agro-ecosystems. However, this benefit has often been overlooked in the design of previous field studies, leaving the net impact of neonicotinoid treated crops on bees relatively unknown. Here, we determine the combined benefit (forage) and cost (insecticide) of oilseed rape grown from thiamethoxam-treated seeds on Bombus terrestris and Apis mellifera colonies. In April 2014, 36 colonies per species were located adjacent to three large oilseed rape fields (12 colonies per field). Another 36 were in three nearby locations in the same agro-ecosystem, but several kilometers distant from any oilseed rape fields. We found that Bombus colony growth and reproduction were unaffected by location (distant versus adjacent) following the two month flowering period. Apis colony and queen survival were unaffected. However, there was a small, but significant, negative relationship between honey and pollen neonicotinoid contamination and Apis colony weight gain. We hypothesize that any sublethal effects of neonicotinoid seed dressings on Bombus colonies are potentially offset by the additional foraging resources provided. A better understanding of the ecological and agronomic factors underlying neonicotinoid residues is needed to inform evidence-based policy