Long-chain polyunsaturated fatty acids (LC-PUFA), particularly arachidonic acid (ARA) and docosahexaenoic acid (DHA), are considered critical for the development of infants and are commonly supplemented in infant formulae. In this study, two common sources of n-3 LC-PUFA, fish oil (FO) and DHA-rich microalgal oil (DMO), were fed to rat pups of mildly n-3 PUFA-deficient dams to compare changes in LC-PUFA of tissue phospholipids. The milk from dams fed a n-3 PUFA-deficient diet contained less n-3 LC-PUFA than that of dams fed a control diet (AIN-93G). The pups' were given orally 1 mg/g weight of either FO or DMO for 17 days between the ages of 5 and 21 days, the pups were weaned, and sacrificed 1 week later for analysis of fatty acid compositions of brain, heart, kidney, spleen, and thymus phospholipids. Although both FO and DMO brought about a recovery in the tissue DHA levels compared to those of the control group (pups from AIN-93G-fed dams), DMO was more effective at restoring tissue LC-PUFA status because it was richer in DHA than FO. FO had a slightly lower PUFA level than that required to bring the LC-PUFA status completely to normal levels in this experiment, and EPA did not accumulate in tissues under the conditions tested here. These results demonstrate the effectiveness of ingesting either FO or DMO in the pre-weaning period for improving mild n-3 PUFA deficiency.