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Standard methods for behavioural studies of Apis mellifera

Summary:

In this BEEBOOK paper we present a set of established methods for quantifying honey bee behaviour. We start with general methods for preparing bees for behavioural assays. Then we introduce assays for quantifying sensory responsiveness to gustatory, visual and olfactory stimuli. Presentation of more complex behaviours like appetitive and aversive learning under controlled laboratory conditions and learning paradigms under free-flying conditions will allow the reader to investigate a large range of cognitive skills in honey bees. Honey bees are very sensitive to changing temperatures. We therefore present experiments which aim at analysing honey bee locomotion in temperature gradients. The complex flight behaviour of honey bees can be investigated under controlled conditions in the laboratory or with sophisticated technologies like harmonic radar or RFID in the field. These methods will be explained in detail in different sections. Honey bees are model organisms in behavioural biology for their complex yet plastic division of labour. To observe the daily behaviour of individual bees in a colony, classical observation hives are very useful. The setting up and use of typical observation hives will be the focus of another section. The honey bee dance language has important characteristics of a real language and has been the focus of numerous studies. We here discuss the background of the honey bee dance language and describe how it can be studied. Finally, the mating of a honey bee queen with drones is essential to survival of the entire colony. We here give detailed and structured information how the mating behaviour of drones and queens can be observed and experimentally manipulated. The ultimate goal of this chapter is to provide the reader with a comprehensive set of experimental protocols for detailed studies on all aspects of honey bee behaviour including investigation of pesticide and insecticide effects.

    1. 2.1. Capturing free-flying bees
    2. 2.1.1. How to catch leaving bees
    3. 2.1.2. How to catch returning foragers
    4. 2.2. Narcotizing bees
    5. 2.3. Marking individual bees
    6. 2.4. Harnessing individual bees
    1. 3.1. Introduction
    2. 3.2. Gustatory responsiveness
    3. 3.3. Responsiveness to light (phototaxis)
    4. 3.4. Responsiveness to odours
    1. 5.1. Classical conditioning
    2. 5.2. Differential olfactory conditioning
    3. 5.3. Tactile conditioning
    4. 5.4. Mechanosensory conditioning
    5. 5.5. Pitfalls
    1. 6.1. Introduction
    2. 6.2. Aversive conditioning in a shuttle box
    3. 6.2.1. Example protocols
    4. 6.2.2. The study of “emotions???
    5. 6.2.3. Standard two-way shuttle box construction
    6. 6.2.4. One-way shuttle box
    7. 6.3. Aversive conditioning in harnessed bees
    8. 6.3.1. Aversive conditioning of proboscis extension
    9. 6.3.2. Aversive conditioning of sting extension
    10. 6.4. Aversive conditioning in free-flying bees
    11. 6.5. Training variables and aversive stimuli
    12. 6.5.1. Electric shock as an aversive stimulus
    13. 6.5.2. Nonstandard aversive stimuli
    14. 6.6. Perspectives for aversive conditioning
    1. 7.1. Introduction
    2. 7.1.1. Temperature and thermotactic orientation in a honey bee hive
    3. 7.1.2. Investigating thermotactic behaviour
    4. 7.1.3. Emulation and application of two-dimensional temperature fields
    5. 7.2. Arena construction
    6. 7.3. Generating a thermal gradient
    7. 7.4. Data analysis
    8. 7.5. Application of thermal gradients
    9. 7.6. Advantages and disadvantages of the method
    1. 8.1. Introduction
    2. 8.2. Roundabout construction
    3. 8.3. Preparation and treatment of bees
    1. 9.1. Introduction
    2. 9.2. Hive setup
    3. 9.3. Behavioural observations
    4. 9.4. Other observation hives
    1. 10.1. Introduction
    2. 10.2. Parameters to consider when studying honey bee dance
    3. 10.3. Measuring dance-related parameters
    4. 10.3.1. Dancer’s behaviour
    5. 10.3.2. Food source location
    6. 10.3.3. To estimate the direction of the food source
    7. 10.3.4. Dance precision
    8. 10.3.5. Dance follower’s behaviour
    9. 10.3.6. Dance attractiveness
    1. 11.1. Recording initial flight path
    2. 11.2. Recording whole flight path
    3. 11.2.1. Radar
    4. 11.2.2. Transponder
    5. 11.2.3. Attaching the transponder
    6. 11.2.4. Field studies
    1. 12.1. Introduction
    2. 12.2. Tracking unmarked bees
    3. 12.3. Tracking individual bees
    4. 12.4. Advanced methods: radio frequency identification
    5. 12.4.1. Equipment
    6. 12.4.2. Location of scanners
    7. 12.4.3. Attaching the RFID tags to the thorax of a bee
    8. 12.4.4. Monitoring activity
    9. 12.4.4.1. Life-long monitoring
    10. 12.4.4.2. Monitoring foraging behaviour
    11. 12.4.5. Parameters of foraging flight
    12. 12.5. Data handling
    13. 12.5.1. Transponder identification
    14. 12.5.2. Data filtering
    15. 12.5.3. Study design
    16. 12.6. Examples of application
    17. 12.6.1. Example 1: Nosema infection for life-long monitoring
    18. 12.6.2. Example 2: Acute effects of sublethal doses of insecticides or acaricides on foraging
    1. 13.1. Introduction
    2. 13.2. Time and weather restrictions for studying mating behaviour
    3. 13.3. Data collection at the flight entrance: daily mating flight period, duration of individual flights, total life time duration and age-dependent flight behaviour
    4. 13.3.1. Determining mating flight periods
    5. 13.3.2. Monitoring individual flight behaviour
    6. 13.3.3. Observing and monitoring queen flights
    7. 13.4. Assembling equipment for the DCA
    8. 13.4.1. Material for balloons and drone traps
    9. 13.4.2. Components for a stable platform in the height of flying drones
    10. 13.4.3. Experiments on copulation
    11. 13.4.4. Drone trap construction
    12. 13.4.5. Catching drones on fly catching paper
    13. 13.5. Starting the experiment
    14. 13.5.1. Balloon preparation
    15. 13.5.2. Locating DCAs
    16. 13.5.3. Experiments with the mast
    1. 14.1. Introduction
    2. 14.2. Training bees to an artificial sucrose feeder in the field
    3. 14.3. Determining sucrose acceptance thresholds in the field
    4. 14.4. Conditioning free-flying bees in the field
    5. 14.4.1. Preparing the bees
    6. 14.4.2. Conditioning the bees
    7. 14.5. Conclusions