Pressure,O2, and CO2, in aquatic Closed Ecological Systems

Pressure increased during net photosynthetic O₂ production in the light and decreased during respiratory O₂ uptake during the dark in aquatic Closed Ecological Systems (CESs) with small head gas volumes. Because most CO₂ will be in the liquid phase as bicarbonate and carbonate anions, and CO₂ is more soluble than O₂, volumes of gaseous CO₂ and gaseous O₂ will not change in a compensatory manner, leading to the development of pressure. Pressure increases were greatest with nutrient rich medium with NaHCO₃ as the carbon source. With more dilute media, pressure was greatest with NaHCO₃, and less with cellulose or no-added carbon. Without adequate turbulence, pressure measurements lagged dissolved O₂ concentrations by several hours and dark respiration would have been especially underestimated in our systems (250–1000 ml). With adequate turbulence (rotary shaker), pressure measurements and dissolved O₂ concentrations generally agreed during lights on/off cycles, but O₂ measurements provided more detail. At 20 °C, 29.9 times as much O₂ will distribute into the gas phase as in the liquid, per unit volume, as a result of the limited solubility of O₂ in water and according to Henry’s Law. Thus even a small head gas volume can contain more O₂ than a larger volume of water. When both dissolved and gaseous O₂ and CO₂ are summed, the changes in Total O₂ and CO₂ are in relatively close agreement when NaHCO₃ is the carbon source. These findings disprove an assumption made in some of Taub’s earlier research that aquatic CESs would remain at approximately atmospheric pressure because approximately equal molar quantities of O₂ and CO₂ would exchange during photosynthesis and respiration; this assumption neglected the distribution of O₂ between water and gas phases. High pressures can occur when NaHCO₃ is the carbon source in nutrient rich media and if head-gas volumes are small relative to the liquid volume; e.g., one “worse case” condition developed 800 mm Hg above atmospheric pressure and broke the glass container. Plastic screw cap closures are likely to leak at high pressures and should not be assumed to seal unless tested at appropriate pressures. Pressure can be reduced by having larger head-gas volumes and using less concentrated nutrient solutions. It is important that pressure changes be considered for both safety and closure, and if total O₂ is used as the measure of net photosynthesis and respiration, the O₂ in the gas phase must be added to the dissolved O₂.     

Community metabolism of aquatic Closed Ecological Systems: Effects of nitrogen sources

To investigate the effect of nitrogen sources on Closed Ecological Systems (CESs), three nitrogen sources (NaNO₃, sodium nitrate; NH₄Cl, ammonium chloride; and NH₄NO₃, ammonium nitrate) were each tested in freshwater CESs consisting of a chemically defined medium, three species of green algae (Ankistrodesmus, Scenedesmus, and Selenastrum), the grazer Daphnia magna, and associated microbes, under 12 h light/12 h dark cycles. It had been hypothesized that the development of high pH in earlier CESs was the result of nitrate utilization, and that ammonium might result in acid conditions, while ammonium nitrate might result in more moderate pH. The three nitrogen sources supported similar densities of algae (estimated by in vivo fluorescence) and similar Daphnia populations. The experiments showed that pH levels rapidly increased when grazers were absent or at low abundances irrespective of the nitrogen source. Consequently, it is hypothesized that carbon cycles, rather than nitrogen sources, are responsible for the pH dynamics. Oxygen diurnal (light:dark) cycles tended to come into balance more quickly than pH. It may be more feasible to convert O₂ data to energy units (using “oxycalorific” values) than CO₂ data since CO₂ dynamics may include other chemical reactions than just photosynthesis and respiration. The feasibility of sustaining grazer populations for at least several weeks in small, simple CESs was demonstrated, along with the ability to monitor algae-grazer dynamics, and the recording of O₂ and pH measurements.