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Biodiversity impacts are rarely included in systems analyses of livestock production. We piloted two approaches toward quantifying biodiversity impacts, pressure‐state‐response (PSR) and potential species loss (PSL), at a cow–calf operation in Florida for which extensive environmental data were available. Using these approaches, we compared livestock production on two vegetation types, semi‐native pasture (SNP) and improved pasture (IMP), and we found fewer deleterious effects on biodiversity associated with SNP (characterized by low stocking rates and no fertilizer) than with IMP, as evidenced by a lower PSL and greater biotic integrity under PSR. Both approaches agreed in the direction of the outcome, but we argue that, when possible, they should be applied complementarily to inform both absolute and per‐unit product biodiversity impacts of livestock production. This research demonstrates how to incorporate biodiversity into life‐cycle perspectives of livestock sustainability assessments when data availability varies, supporting the expansion of multi‐metric, holistic evaluations that are absent from most livestock system analyses. [ABSTRACT FROM AUTHOR]