Elemental sulfur (S 0 ) is associated with many geochemically diverse hot springs, yet little is known about the phylogeny, physiology, and ecology of the organisms involved in its cycling. Here we report the isolation, characterization, and ecology of two novel, S 0 -reducing Crenarchaea from an acid geothermal spring referred to as Dragon Spring. Isolate 18U65 grows optimally at 70 to 72°C and at pH 2.5 to 3.0, while isolate 18D70 grows optimally at 81°C and pH 3.0. Both isolates are chemoorganotrophs, dependent on complex peptidecontaining carbon sources, S 0 , and anaerobic conditions for respiration-dependent growth. Glycerol dialkyl glycerol tetraethers (GDGTs) containing four to six cyclopentyl rings were present in the lipid fraction of isolates 18U65 and 18D70. Physiological characterization suggests that the isolates are adapted to the physicochemical conditions of Dragon Spring and can utilize the natural organic matter in the spring as a carbon and energy source. Quantitative PCR analysis of 16S rRNA genes associated with the S 0 flocs recovered from several acid geothermal springs using isolate-specific primers indicates that these two populations together represent 17 to 37% of the floc-associated DNA. The physiological characteristics of isolates 18U65 and 18D70 are consistent with their potential widespread distribution and putative role in the cycling of sulfur in acid geothermal springs throughout the Yellowstone National Park geothermal complex. Based on phenotypic and genetic characterization, the designations Caldisphaera draconis sp. nov. and Acidilobus sulfurireducens sp. nov. are proposed for isolates 18U65 and 18D70, respectively.

Indigenous drinking water organisms were used to establish nitrification in glass reactors containing copper or polyvinyl chloride (PVC) surfaces. The reactors were fed soil-derived humics as the organic carbon source and ammonium sulfate as the nitrogen source in biologically treated tap water. Water in the reactors was stagnant for 8 h and then flowed for 5 min to simulate conditions in household plumbing. Following the establishment of complete nitrification (conversion of ammonia to nitrate) in both reactor types, various inhibitors of nitrification were tested followed by a period where recovery of nitrification was observed. In one PVC reactor, copper was gradually introduced up to 1.3 ppm. To ensure that most of the copper was in the ionic form, the pH of the influent was then gradually lowered to 6.6. No significant change in nitrification was observed in the presence of copper. Chlorite was introduced into copper and PVC reactors at doses increasing from 0.2 ppm to 20 ppm. There was limited effect on the PVC system and inhibition in the copper reactor only at 20 ppm. Chloramine was tested at chlorine to ammonia ratios ranging from 0.5:1 to 5:1. Nitrification activity was impacted significantly at a 5:1 ratio and ultimately stopped, with the fastest response being in the copper system. Whenever a control mechanism was tested, there was increased release of copper from the reactors with copper coupons. In all cases, nitrification recovered when inhibitors were removed but the rates of recovery differed depending on the treatment method and coupon surface.