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OBJECTIVES: This study was designed to evaluate the activities of liver enzymes in a cohort of HIV-positive pregnant women on highly active antiretroviral therapy (HAART). METHODS: The activities of liver enzymes-ALP, ALT and AST- were assessed in ninety pregnant subjects aged 20-40 years at University of Nigeria Teaching Hospital (UNTH) Ituku-Ozalla Enugu. Thirty of the subjects were apparently healthy HIV seronegative pregnant women that served as the negative control; while sixty of the subjects were confirmed HIV infected subjects, thirty of which were on HAART (Test subjects); while thirty were not on HAART (positive control) and were further grouped into trimesters. Serum samples were prepared from blood samples collected from the subjects and assayed using standard methods. RESULTS: The ALP, ALT and AST activities of the test subjects where compared with negative control in the first, second and third trimesters which showed significant increases in ALP (p<0.001,p<0.05 and p<0.05 respectively); ALT in all the trimesters were significantly different (p<0.001); while in AST significant differences could be observed in third trimesters p<0.05). When compared with positive control, ALP showed significant increase in all trimesters (p<0.001, p<0.05, and p<0.001). ALT was slightly but not significantly increased (p>0.05) in the second trimester, but significantly increased in first and third trimesters (p<0.001) but had no significant difference in the second trimester (p>0.05). The comparisons of negative and positive controls showed differences in enzyme activities in each of the trimesters. CONCLUSION: Pregnancy, HIV infection, and drug treatment appear to have a cumulative effect though the treatment group had the highest level of increased ALT, AST and ALP activity across the trimesters.