Changes in milk composition are described for three species of free-ranging insectivorous bats (Myotis lucifugus, M. velifer, and Tadarida brasiliensis) from early to mid (peak) lactation. Dry matter and energy concentrations in milk increased from early to mid-lactation. In M. lucifugus and T. brasiliensis, but not M. velifer, these increases were due largely to a rise in fat concentration, since protein and carbohydrate remained relatively constant. Energy content of milk (kJ.g-1) for each species from early through mid-lactation was related to dry matter (DM) as follows: M. lucifugus (y = 0.31 DM-0.32, r2 = 0.68), M. velifer (y = 0.48 DM-5.08, r2 = 0.99), and T. brasiliensis (y = 0.37 DM-1.51, r2 = 0.61). Comparison of the effect of sampling method on milk composition of T. brasiliensis indicated that fat, dry matter, and energy concentrations increased significantly from pre-dawn to pre-noon samples. Relatively high fat and low water levels in T. brasiliensis milk may reflect the limited access that lactating females have to free water, as well as need to minimize mass of stored milk during long foraging trips. Conversely, lower fat concentrations and higher water levels in milk in M. lucifugus and M. velifer may relate to the propensity for colonies of these two species to roost and forage near bodies of water. In addition, differences in milk fat concentrations observed among the three species may correlate to daily suckling schedules.(ABSTRACT TRUNCATED AT 250 WORDS)