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Abstract Despite the ecological significance of desert springs, little is known about relationships between spring hydrogeochemistry and ecology, particularly over multiple trophic levels. Here, we surveyed microbial communities (bacteria and archaea) and benthic macroinvertebrate (BMI) communities in springs that recharge through granitoid bedrock in Owens Valley, CA, to determine whether subtle geochemical differences imparted by metamorphosed roof pendant weathering in the source area affect spring ecosystems. Relative to other springs, roof‐pendant‐recharged springs were characterized by elevated (1) Ca2+/Na+, Ca2+/Mg2+, and divalent/monovalent cation ratios, (2) relative abundance of taxa inferred to be benthic aerobes and prosthecate/stalked bacteria, and (3) abundance and diversity of shredder and collector gatherer BMIs. These BMI feeding groups graze on biofilm communities and stimulate bacterial degradation of particulate organic matter, consistent with extensive bacteria–BMI connections in co‐occurrence networks in these springs. Springs not impacted by roof pendants were instead enriched with taxa inferred to be anaerobes, chemolithotrophs, and low‐diversity BMI communities with poor bacteria–BMI network connectivity. We speculate that excess Ca2+ derived from roof pendant weathering plays a key role in biofilm formation on coarse granite substrates, with subsequent trophic interactions between benthic microbial biofilms and diverse BMI shredders/collector gatherers.