Stromatolites are abundant and widely distributed within two narrow stratigraphic intervals in Middle (Darriwilian) and Upper (lower Turinian) Ordovician strata of the Laurentian foreland interior in the Ottawa Embayment, eastern Ontario. These lithostratigraphic markers coincide with rapid, tectonically driven flooding of the foreland interior and may identify an opportunistic microbial response to nutrient loading with shallow (peritidal, subtidal) marine reworking of terrestrial or nutrient-rich coastal systems. The remaining (Chatfieldian–Edenian) foreland-platform succession represents deeper-water deposition in response to elevated subsidence rates along the Laurentian margin. Stromatolites are absent in this higher-energy setting, but microbial calcite is preserved as coccoid-like microencrustations on skeletal debris. Stromatolites of Darriwilian age are dolomitic, occur in the Carillon Formation, and are part of a regional (200+ km) onlap of peritidal sediment during onset of Taconic orogenesis. Stratiform to large (2 m diameter) low-relief domal stromatolites contain rhythmic laminations of inclusion-rich and -poor dolomicrospar grouped by Fe-oxide-stained erosional surfaces. Domal forms also contain thrombolitic microstructure. Patterns of lamination and stable (C, O) isotopes suggest a balance between abiotic and microbial carbonate production, likely influenced by water depth and temperature. Stromatolites of Turinian age are calcitic and form a regional (80+ km) thin (<8 m) subtidal biostromal unit in the lowermost Pamelia Formation. Their occurrence defines an abrupt vertical transition from initial intrabasinal transgressive subtidal phosphatic siliciclastics and carbonates to an interbasinal stratigraphy of peritidal lime mudstone. Arrhythmic laminations of microbial peloid packstone, with possible eukaryote alga moulds, and locally evaporitic spongiostromate microstructure identify salinity and energy as primary depositional controls. The stromatolite–?eukaryotic association is similar to some modern subtidal microstructures and is part of the reef-community diversification in the Ordovician.