Social isolation of ants in queenless, broodless conditions

The aim of this protocol is to alter the social environment of ants and observe the effects of this treatment on components of trophallactic fluid. 

Protocol

1. Select N colony replicates.

2. Prepare N fluoned boxes with shelter, water and insect-free food (maple syrup, chicken eggs, tuna and agar).

3. Prepare trophallactic fluid collection materials (capillaries, CO2, N*2 eppendorf tubes, etc.).

4. Collect ~ 75% in-nest workers (nurse-like) and 25% out-of-nest workers (forager-like) to approximate a colony's natural ratio. 

1. For groups of 25–30 C. floridanus worker ants, collect with forceps five to eight workers from the foraging arena and place in a temporary fluoned box and then collect 20-25 individuals from the nest area under the nest lid on or near brood as quickly as possible (ants will scatter when nest is opened). 

5. Collect TF fluid under CO2 anesthesia and place recovering ants into prepared fluoned boxes with shelter, food, etc. 

6. Repeat steps 4 and 5 for each replicate. 

7. Feed all replicates after one week.

8. Two weeks after first TF collecton, repeat step 5 for remaining individuals in each grouped isolation. Note that some death is expected even in grouped isolation (Koto et al. 2014). 

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