Why did carbon become the pseudo-limiting factor in aquatic closed ecological systems?

Closure from the earth's atmosphere is a critical test of an ecosystem's ability to function. In our earlier testing of autotrophic Closed Ecological Systems (CESs), a C:N ratio of 26.4 (3.3 mM NaHCO₃ and 0.125 mM NaNO3) supported algal and Daphnia populations for months, but developed extreme pH values (∼11 ungrazed, >10, grazed), suggesting that the systems were carbon-limited. Only approximately half the HCO₃^- (bicarbonate) would be expected to be available to green algae, the other portion becoming CO₃^-2 (carbonate). In an experiment described here, CESs were developed to explore a greater range of C:N ratios. To keep the medium from becoming too osmotically concentrated, NaNO₃ was reduced to 0.0312 mM and NaHCO₃ tested at 3.3, 13.2, and 26.4 mM, resulting in nominal C:N ratios of 105, 422, and 845. However, additional carbon was not beneficial to long-term survival of the organisms. The algal abundance was relatively insensitive to C:N ratio; greater concentrations of C were not beneficial. Daphnia populations were sensitive to C:N ratio and persisted longer at the lowest C:N ratio of 105. All of the C:N ratios tested in these CESs are outside of the expected range suggested from ecological studies, which is based on the Redfield Ratio of 6.625 C:N, the expected chemical composition of algae. Two potential explanations for the apparent high C demand in our CESs are suggested by the literature. The first is production of fatty algal cells, e.g., one of the algal species, Scenedesmus obliquus, is reported to produce high-lipid cells that could have a higher C:N ratio than the Redfield Ratio. The second is "carbon overconsumption," which has been suggested for N-limited marine phytoplankton communities dominated by diatoms or nutrient deficient algal communities dominated by small cells that are under-represented by chlorophyll a measurements. The unexpected C dynamics found in our CES tests could be relevant to the design of biological life support systems that must be provisioned with adequate elements for long-term ecosystem functionality. If the actual demand for C is underestimated, its storage may be inadequate.