Temporal niche partitioning can be a viable mechanism for coexistence, but has received less attention than other niche axes. We characterized and compared patterns of activity, and overlap of temporal activity among the five common rodent species from a tropical semideciduous forest (TSF) and between the two common rodent species from cloud forest (CF) at El Cielo Biosphere Reserve in Mexico. Capture frequencies over 2-h intervals, obtained via live trapping (6850 trap-nights) in chosen months over 3 y formed the empirical basis for analyses. Trap transects were set from 19h00 to 07h00 and checked every 2 h. Analyses of 484 captures evinced two distinct assemblages. The TSF assemblage was diverse and with non-random temporal niche segregation, whereas the CF assemblage was depauperate with its two dominant species evincing the same activity pattern. Predator avoidance between open- and closed-microhabitat species, as well as niche complementarity may explain temporal segregation at TSF. This is the first documentation of assemblage-wide non-random temporal segregation of neotropical rodents. Time of activity may be a largely under-appreciated mechanism in other species-rich tropical rodent assemblages as well as in other species-rich biotas.