Rotational grazing is sometimes promoted for grassland bird conservation, but the benefits to wildlife have not been comprehensively documented. We examined effects of twice-over rotational grazing on nesting success of grassland songbirds in southwestern Manitoba, Canada in comparison to season-long grazing. We monitored nesting attempts and collected structural vegetation data in 2011 (during a 1/300 flood event), and 2012 (average water levels), for five species of obligate grassland bird species (n = 110) and one shrub-nesting species (n = 41).Nesting analyses were conducted using logistic exposure models. Nesting success was 2.4 to 4 times lower in twice-over grazed pastures compared with season-long grazed pastures, perhaps because of the increased cattle density during the short grazing periods of the twice-over system. Nests protected by shrubs from grazing activities of cattle did not show this pattern. The grazing system did not have an effect on vegetation structure. This suggests that twice-over rotational grazing does not benefit grassland songbirds in northern mixed-grass prairies, and that caution must be taken before implementing this grazing system in areas intended to promote biodiversity conservation.