With the use of continuous culture systems, rates of dinitrogen (N-2) and carbon (C) fixation and nitrogen (N)- and C-based doubling times were assessed in Trichodesmium IMS101 growing exponentially at steady state dilution rates of 0.10, 0.20, and 0.33 d(-1) (doubling times of 10, 5, and 3 d - within the range reported for natural populations). Rates of C fixation, N-2 fixation, and N release were examined in replicate culture systems with several techniques. Biomass-specific C uptake varied little with population doubling time, but N-2 fixation and N release varied markedly among treatments. Total daily gross N-2 fixation rates and estimated N release rates were higher in cultures with higher dilution rates. Cultures grown at lower dilution rates had higher daily C:N, fixation ratios and lower N release rates. Consistent with other studies, it was estimated that Trichodesmium released about 80-90% of their recently fixed N-2 during growth. Turnover of cellular C estimated from carbon fixation was a good estimator of population growth rates in steady state cultures, whereas turnover of cellular N estimated from gross or net N, fixation was not. Small changes in temperature (24 degrees C vs. 28 degrees C) did not appear to affect gross N2 fixation, whereas inorganic phosphorus (1 vs. 5 mu mol L-1) supply had a large effect on N-2 fixation. These results suggest that continuous culture systems are excellent for elucidating physiological responses of Trichodesmium under ecologically relevant growth conditions and provide a framework for assessing highly variable field estimates of N-2 and C fixation.