The Chamberlain Formation, one of the lower members of the early Mesoproterozoic Belt Supergroup, has previously yielded low-diversity assemblages of microfossils but the reported fossils were of limited utility for inferring paleoenvironmental conditions. Here, we describe substantially more diverse microfossil assemblages from drill core of the Chamberlain Formation obtained from the Black Butte mine locality near White Sulphur Springs, Montana. The Chamberlain Formation biota contains abundant Valeria, Leiosphaeridia, Synsphaeridium, and Lineaforma, with lesser amounts of Satka, Symplassosphaeridium, andConiunctiophycus. The assemblages partially overlap with, but are distinct from, microfossils recently reported from the Greyson Formation, another unit from the Helena embayment of the Belt Supergroup. Since the overlapping taxa exhibit similar states of preservation but dissimilar relative abundances, we interpret the assemblages as reflective of distinct paleoenvironmental conditions of the sampled sections of the Chamberlain and Greyson Formations. The Chamberlain Formation assemblages are most comparable to microfossil groupings reported from the Bylot Supergroup of Canada and the Roper Group of Australia from sediments from very shallow-water (supratidal to lower shoreface) marine environments. This comparison corroborates previous hypotheses on the basis of sedimentological data that the lower Chamberlain Formation sediments were formed in a lagoonal or mud-flat environment. By contrast, the Greyson Formation assemblages are most comparable to microfossil groupings associated with sediments from shallow-shelf marine environments. The fidelity of comparisons among the 1.2 Ga Bylot Supergroup, 1.49 Ga Roper Group, and 1.45 Ga Belt Supergroup assemblages indicates that the groups of microorganisms that produced these assemblages, and their associations with the paleoenvironments that they inhabited, may have been characteristic of the littoral marine biosphere throughout much of the Mesoproterozoic.