Hemolymph was pooled from five to eight larvae to obtain 1 μl for assay. Glucose was measured by adding to 99 μl of Thermo Infinity Glucose Reagent (TR15321) frozen in a 96-well plate, then thawed to allow the detection reactions to occur simultaneously for all wells, and processed as per the manufacturer’s instructions. The level of trehalose was measured using the same reagent after digestion with trehalase, with a ten-fold dilution because trehalose levels are higher than those of glucose. 1 μl of hemolymph was incubated in 25 μl 0.25 M sodium carbonate at 95°C for 2 hours in a thermal cycler, cooled to room temperature, and 8 μl of 1 M acetic acid and 66 μl of 0.25 M sodium acetate (pH 5.2) were added to make a digestion buffer. 1 μl of porcine trehalase (Sigma T8778) was added to 40 μl of this mixture and incubated at 37°C overnight. The resulting glucose was analyzed using 10 μl reaction + 90 μl Infinity reagent as above. Glucose and trehalose standards were treated simultaneously and used to quantify the sugar levels in hemolymph.