Experimental design and sample distribution.

<p>(A) Field collected eggs were brought into the laboratory. Once they reached the free feeding stage (Gonser stage 25), 140 tadpoles were individually raised in 3 L plastic containers until Gosner stage 35. (B) As tadpoles reached stage 35, they were assigned to either constant water volume (3L), or reduced water volume (350 mL). (C) Tadpoles were then raised up to predetermined stages: Gosner stages 38 and 42 (stages at which the physiological parameters were determined), or until metamorphosis was complete (Gosner stage 46). (D) Corticosterone (CORT) was determined by electroimmuno assays from plasma samples obtained from 80 tadpoles across developmental stages and water treatments. Tail tissue from those tadpoles was used for thyroid hormone (TH) radioimmuno assays and determination of the level of expression of the thyroid hormone receptor TRβ via qPCR. Levels of oxidative stress and activity of antioxidant enzymes were determined after whole body homogenization.</p>