This study evaluated the effects of 1) increasing the concentration of sulfate in drinking water on dry matter intake (DMI), water intake, ruminal fermentation, and apparent total-tract digestibility; and 2) water sulfate concentration and bismuth subsalicylate (BSS) dose on in vitro ruminal fermentation. Eight ruminally cannulated beef heifers (382 ± 45 kg) were used in a replicated incomplete 3 × 3 Latin square design. Water treatments contained 342 ± 29 (LS), 2,785 ± 72 (MS), or 4,948 ± 163 mg/L (HS) sulfate. Ruminal digesta was collected to evaluate water sulfate and BSS dose (0.0, 0.2, 0.4, and 0.6% dry matter) on ruminal H2S production in vitro. Water intake increased linearly as water sulfate concentration increased (P = 0.002) but DMI was not affected. Heifers drinking MS and HS had greater ruminal H2S at 10.5 h after water provision than LS (sulfate × time, P < 0.001). In vitro H2S production (µg and µg/mL of rumen inoculum) increased and plateaued with increasing sulfate (P < 0.001) and was linearly reduced (P < 0.001) by increasing BSS dose. Increasing water sulfate concentrations did not negatively affect water or feed intake but increased ruminal H2S concentrations. Bismuth subsalicylate may reduce H2S production.