Standard methods for behavioural studies of Apis mellifera.

Authors: Ricarda Scheiner, Charles I Abramson, Robert Brodschneider, Karl Crailsheim, Walter M Farina, Stefan Fuchs, Bernd Grünewald, Sybille Hahshold, Marlene Karrer, Gudrun Koeniger, Niko Koeniger, Randolf Menzel, Samir Mujagic, Gerald Radspieler, Thomas Schmickl, Christof Schneider, Adam J Siegel, Martina Szopek, Ronald Thenius.

Table of contents

1. General introduction
2. Methods for preparing bees for behavioural tests
   2.1. Capturing free-flying bees

      2.1.1. How to catch leaving bees
      2.1.2. How to catch returning foragers
   2.2. Narcotizing bees
   2.3. Marking individual bees
   2.4. Harnessing individual bees

3. Quantifying sensory responsiveness
   3.1. Introduction
   3.2. Gustatory responsiveness
   3.3. Responsiveness to light (phototaxis)
   3.4. Responsiveness to odours
4. Quantifying non-associative learning
5. Quantifying associative appetitive learning and memory in the laboratory

   5.1. Classical conditioning
   5.2. Differential olfactory conditioning
   5.3. Tactile conditioning
   5.4. Mechanosensory conditioning
   5.5. Pitfalls
6. Quantifying associative aversive learning in the laboratory
   6.1. Introduction
   6.2. Aversive conditioning in a shuttle box

      6.2.1. Example protocols
      6.2.2. The study of “emotions”
      6.2.3. Standard two-way shuttle box construction
      6.2.4. One-way shuttle box
   6.3. Aversive conditioning in harnessed bees
      6.3.1. Aversive conditioning of proboscis extension
      6.3.2. Aversive conditioning of sting extension
   6.4. Aversive conditioning in free-flying bees
   6.5. Training variables and aversive stimuli

      6.5.1. Electric shock as an aversive stimulus
      6.5.2. Nonstandard aversive stimuli
   6.6. Perspectives for aversive conditioning
7. Investigating honey bee locomotion in complex and dynamic temperature gradient fields
   7.1. Introduction

      7.1.1. Temperature and thermotactic orientation in a honey bee hive
      7.1.2. Investigating thermotactic behaviour
      7.1.3. Emulation and application of two-dimensional temperature fields
   7.2 Arena construction
   7.3. Generating a thermal gradient
   7.4. Data analysis
   7.5. Application of thermal gradients
   7.6. Advantages and disadvantages of the method
8. Testing honey bee flight capability in the laboratory

   8.1. Introduction
   8.2. Roundabout construction
   8.3. Preparation and treatment of bees
9. Working with honey bee observation hives
   9.1. Introduction
   9.2. Hive setup
   9.3. Behavioural observations
   9.4. Other observation hives
10. The honey bee dance: background, knowledge and observation
   10.1. Introduction
   10.2 Parameters to consider when studying honey bee dance
   10.3 Measuring dance-related parameters

      10.3.1 Dancer's behaviour
      10.3.2 Food source location
      10.3.3 To estimate the direction of the food source:
      10.3.4 Dance precision
      10.3.5 Dance follower's behaviour
      10.3.6 Dance attractiveness
   11. Honey bee navigation: tracking bees with harmonic radar
      11.1. Recording initial flight path
      11.2. Recording whole flight path

         11.2.1. Radar
         11.2.2. Transponder
         11.2.3. Attaching the transponder
         11.2.4. Field studies
12. Monitoring honey bee flight – BeeScan and RFID technologies
   12.1. Introduction
   12.2. Tracking unmarked bees
   12.3. Tracking individual bees
   12.4. Advanced methods: radio frequency identification

      12.4.1. Equipment
      12.4.2. Location of scanners
      12.4.3. Attaching the RFID tags to the thorax of a bee
      12.4.4. Monitoring activity
      12.4.5. Parameters of foraging flight
   12.5. Data handling
      12.5.1. Transponder identification
      12.5.2. Data filtering
      12.5.3. Study design
   12.6. Examples of application
      12.6.1. Example 1: Nosema infection for life-long monitoring
      12.6.2. Example 2: Acute effects of sublethal doses of insecticides or acaricides on foraging
13. Equipment and devices for experiments on mating behaviour
   13.1. Introduction
   13.2. Time and weather restrictions for studying mating behaviour
   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

      13.3.1. Determining mating flight periods
      13.3.2. Monitoring individual flight behaviour
      13.3.3. Observing and monitoring queen flights
   13.4. Assembling equipment for the DCA
      13.4.1. Material for balloons and drone traps
      13.4.2. Components for a stable platform in the height of flying drones
      13.4.3. Experiments on copulation
      13.4.4. Drone trap construction
      13.4.5. Catching drones on fly catching paper
   13.5 Starting the experiment
      13.5.1. Balloon preparation
      13.5.2. Locating DCAs
      13.5.3 Experiments with the mast
14. Appetitive learning in the field
   14.1. Introduction
   14.2. Training bees to an artificial sucrose feeder in the field
   14.3. Determining sucrose acceptance thresholds in the field
   14.4. Conditioning free-flying bees in the field

      14.4.1. Preparing the bees
      14.4.2. Conditioning the bees
   14.5. Conclusions
15. General conclusions
16. Acknowledgements
17. References