A Quiet Riot: Furthering the discussion on aerobic heterotrophy in deep sediments

North Pond, a sediment deposit ringed by basalt outcrops just west of the Mid-Atlantic Ridge, remains a site of intense study of the subseafloor biosphere. During IODP Expedition 336, core samples of sediment and basalt were drilled and permanent CORK observatories were installed in the basalt crust. Heterotrophic enrichments were started aboard ship and multiple aerobic, heterotrophic bacterial isolates were obtained from two sediment horizons. Isolate identities were compared to sequences from drilling fluid and surrounding sediment to establish the likelihood of their sedimentary source. Three isolates currently in pure culture are from site U1382B and include an Arthrobacter species from 4 meters below seafloor (mbsf) as well as a Paracoccus and Pseudomonas species from 70 mbsf. All isolates grow at tested temperatures of 4 to 37°C. Only the Arthrobacter species grows at 42°C and no isolates grew at 50°C. The presence of aerobic microorganisms at these depths is consistent with previously published oxygen profiles of site U1382B where O2 is present in low amounts (10 to 20μm) at both 4 mbsf (originating from overlying seawater) and 70 mbsf (originating from subseafloor aquifer leaching into deep sediment), yet substantial enough to support aerobic heterotrophy. Despite similar oxygen concentrations, two key differences between these depths are the origin and quality of organic matter and the surrounding lithology. Section 1H4 from site U1382B, where the Arthrobacter species was isolated, consists primarily of a nanofossil ooze. Section 8H6 (~70 mbsf) is much more clay-rich. Previous explorations of microbial heterotrophy in North Pond sediments using 14C-acetate have suggested that this metabolism may be linked to particular lithologies. A 2011 study noted higher rates of potential aerobic heterotrophy in sandy and clay-rich layers compared to nannofossil ooze layers. Since isolates are from different depths, ages and lithologies they can be used to examine the potential fitness of an indigenous organism to it's surrounding environment, by comparatively studying the shallow and deep isolates in each others habitats.