Effects of hydrostatic pressure on microbial alteration of sinking fecal pellets

We used a new experimental device called PASS (PArticle Sinking Simulator) during MedFlux to simulate changes in in situ hydrostatic pressure that particles experience sinking from mesopelagic to bathypelagic depths. Particles, largely fecal pellets, were collected at 200 m using a settling velocity NetTrap (SV NetTrap) in Ligurian Sea in April 2006 and incubated in high-pressure bottles (HPBs) of the PASS system under both atmospheric and continuously increasing pressure conditions, simulating the pressure change experienced at a sinking rate of 200 m d−1. Chemical changes over time were evaluated by measuring particulate organic carbon (POC), carbohydrates, transparent exopolymer particles (TEP), amino acids, lipids, and chloropigments, as well as dissolved organic carbon (DOC) and dissolved carbohydrates. Microbial changes were evaluated microscopically, using diamidinophenylindole (DAPI) stain for total cell counts and catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) for phylogenetic distinctions. Concentrations (normalized to POC) of particulate chloropigments, carbohydrates and TEP decreased under both sets of incubation conditions, although less under the increasing pressure regime than under atmospheric conditions. By contrast, dissolved carbohydrates (normalized to DOC) were higher after incubation and significantly higher under atmospheric conditions, suggesting they were produced at the expense of the particulate fraction. POC-normalized particulate wax/steryl esters increased only under pressure, suggesting biochemical responses of prokaryotes to the increasing pressure regime. The prokaryotic community initially consisted of 43% Bacteria, 12% Crenarchaea and 11% Euryarchaea. After incubation, Bacteria dominated (∼90%) the prokaryote community in all cases, with γ-Proteobacteria comprising the greatest fraction, followed by the Cytophaga-Flavobacter cluster and α-Proteobacteria group. Using the PASS system, we obtained chemical and microbial evidence that degradation by prokaryotes associated with fecal pellets sinking through mesopelagic waters is limited by the increasing pressure they experience.