Creating Controlled CO2 Perturbation Experiments on the Seafloor - Development of FOCE Techniques

Experimental recent progress on the design and testing of systems for carrying out controlled CO₂ perturbation experiments on the sea floor with the goal of simulating the conditions of a future high CO₂ world. Controlled CO₂ enrichment (FACE) experiments have long been carried out on land to investigate the effects of elevated atmospheric CO₂ levels on vegetation, but only limited work on CO₂ enrichment on enclosed systems has yet been carried out in the ocean. With rising concern over the impacts of ocean acidification on marine life there is a need for greatly improved techniques for carrying out in situ experiments, which can create a DeltapH of 0.3 to 0.5 by addition of CO₂, on natural ecosystems such as coral reefs, cold water corals, and other sensitive benthic habitats. This is no easy task. Unlike land based experiments where simple mixing in air is all that is required, CO₂ has complex chemistry in seawater with significantly slow reaction kinetics. Scientists must design systems to take this into account. The net result of adding a small quantity of CO₂ to sea water is to reduce the concentration of dissolved carbonate ion, and increase bicarbonate ion through the following reaction:CO₂+H₂O+CO₃^2- -> 2HCO₃ In practice the reaction between CO₂ and H₂O is slow and is a complex function of temperature, pH, and TCO₂, with the reaction proceeding more rapidly at lower pH and higher temperatures. Marine animals in the natural ocean will typically experience only small and temporary shifts from environmental CO<sub>2</sub&- gt; equilibrium. Valid perturbation experiments must try to expose an experimental region to a stable lower pH condition, and avoid large and rapid pH variability. The most common sensor used for experimental control is the pH electrode, and this detects only H⁺ ion, not any of the dissolved CO₂ species. We first explored the reaction kinetics of a CO₂ perturbation in a series of closed loop pH cell experiments carried out at various depths under ROV control. These were found to be well matched to the Zeebe & Wolf-Gladrow [1] model. From these results, functions for the delay time required for equilibrium were devised and a design for a delay loop to achieve at least 2 e-folding times between CO₂ injection and animal exposure was developed. We tested this prototype system in October 2007 in a series of ROV controlled experiments at a depth of 1000 meters. The working fluid used for enrichment was surface sea water saturated at one atmosphere with pure CO₂ gas to create a solution of about pH 4.8 and 56 mM total CO₂. This was carried to depth in a 56 liter piston accumulator, and dispensed as needed into a flexible polyethylene bag for subsequent addition into the experimental unit. The design consisted of a 4 meter delay loop leading to a control volume (square box, 25 cm per side) outfitted with three pH electrodes and a CTD. To determine the uniformity of the pH, two pH electrodes were positioned in the control volume and a third electrode was positioned just beyond the control volume in the flow stream. Ambient seawater, pumped at a desired rate with a modified thruster, was mixed at the beginning of the delay loop with controlled continuous injection of the CO₂-rich working fluid in a ratio typically of about 2001 depending on the pH perturbation desired. For these initial tests, a feed-