The Carpentaria province (McArthur basin and Mount Isa inlier) in northern Australia is one of the most important districts for clastic-dominated (CD-type) massive sulfide deposits. The George Fisher Zn-Pb-Ag deposit, located in this province, is hosted by the carbonaceous Urquhart Shale Formation (ca. 1654 Ma) in a region that has an active history of metamorphism and tectonism. In this study, paragenetically constrained pyrite in samples from the George Fisher deposit and unmineralized Urquhart Shale have been analyzed in situ using secondary ion mass spectrometry (SIMS) of sulfur isotopes (δ34S values). Samples were taken from four drill cores through the main orebodies at George Fisher and one drill core through correlative, unmineralized Urquhart Shale (Shovel Flats area). Five generations of pyrite were identified at George Fisher and record a protracted history of sulfate reduction under diagenetic and subsequent hydrothermal conditions: (1) fine-grained, subhedral-spheroidal pyrite (Py-0), (2) coarse-grained, anhedral pyrite (Py-1) associated with ore-stage 1 sphalerite and galena, (3) coarse-grained, euhedral pyrite (Py-2) associated with ore-stage 2 sphalerite, galena, and pyrrhotite, (4) massive subhedral to euhedral pyrite (Py-3) associated with ore-stage 3 chalcopyrite, pyrrhotite, galena, and sphalerite, and (5) coarse-grained euhedral pyrite (Py-euh), which occurs only in unmineralized rocks. In the unmineralized Shovel Flats drill core, only Py-0 and Py-euh are present. Whereas pre-ore pyrite (Py-0) preserves negative δ34S values (–8.1 to 11.8‰), the ore-stage pyrites (Py-1, Py-2, and Py-3) have higher δ34S values (7.8–33.3, 1.9–12.7, and 23.4–28.2‰, respectively). The highest δ34S values (7.2–33.9‰) are preserved in Py-euh. In combination with petrographic observations, the δ34S values of pyrite provide evidence of three different processes responsible for the reduction of sulfate at George Fisher. Reduced sulfur in fine-grained pyrite (Py-0) formed via microbial sulfate reduction (MSR) under open-system conditions prior to the first generation of hydrothermal pyrite (Py-1) in ore-stage 1, which most likely formed via thermochemical sulfate reduction (TSR). During deformation, previously formed sulfide phases were then recycled and replaced during a second hydrothermal event (ore-stage 2), resulting in intermediate sulfur isotope values. Another syndeformational hydrothermal Cu event, involving a sulfate-bearing fluid, formed ore-stage 3 via TSR. This study demonstrates that the fine-grained pyrite formed pre-ore under conditions open to sulfate and outlines the role of multiple stages of sulfide formation in producing high-grade Zn-Pb-Ag orebodies in the Mount Isa inlier.