In order for microalgae to be a cost-effective renewable energy source, a high CO2-transfer efficiency (CTE) is necessary. Using hollow-fiber membranes (HFM), membrane carbonation (MC) in microalgal cultivation can be used to achieve a CTE near 100%. Due to the diurnal cycle in outdoor algal cultivation, an inconsistent CO2 demand with temperature fluctuations can cause pore wetting of the inner and outer fiber layers in composite HFMs. In addition, the presence of supersaturated O2 during high algal growth may change the gas transfer dynamics of the fibers, which can be critical when trying to selectively remove CO2 from a valuable gas such as biogas. This study evaluated fiber performance under conditions that mimic these effects by analyzing the carbon transfer efficiency (CTE), CO2 flux (JCO2), and outlet CO2 concentration compared to baseline values. Wetting of the interior fiber macropores resulted in an average 32% ± 8.3% decrease in flux, which was greater than for flooding of the outer macropores, which showed no significant change. All tests resulted in a decrease in CTE and an increase in outlet CO2. The presence of elevated O2 levels did not decrease the CO2 flux compared to baseline values, but it increased the O2 concentration and decreased the CH4 concentration at the distal end of the fibers. These findings highlight that liquid accumulation can decrease HFM performance during MC for microalgal cultivation, while the presence of supersaturated O2 can reduce separation efficiency.