In the past decades, the African buffalo (Syncerus caffer) population has steadily declined; an estimated reduction of 31.2% over 3 generations has resulted in classifying it as near threatened by the International Union for Conservation of Nature. Therefore, the aim of this work was a preliminary evaluation of the size and number of ovarian follicles and oocyte quality and quantity in African buffaloes in relationship to age, tuberculosis, and pregnancy status to assess feasibility of in vitro embryo production (IVEP) as a tool for conservation and propagation of this species. The study occurred during the winter dry season (Jul–Aug) of 2018 within Huluhluwe‐Imfolozi National Park in Kwa‐Zulu Natal, South Africa during the routine culling procedures of African buffalo carried out at the park by the provincial authorities as means of disease management and population control. We obtained ovaries from 39 adults and 10 juveniles, and transported them to the field laboratory where we counted follicles, collected oocytes by aspiration and slicing, and classified oocytes according to their morphology. Adult animals had more small and medium follicles compared to juveniles (small: 14.5 ± 0.1 [SE] vs. 14.0 ± 0.1; medium: 4.6 ± 0.7 vs. 2.0 ± 1.3); however, juvenile animals had more cumulus‐oocyte complexes (COCs). Pregnancy and tuberculosis status did not affect COC recovery rate and quality. The oocyte recovery rate is comparable to cattle and higher than in water buffalo (Bubalus bubalis). Therefore, our results suggest that IVEP could be an effective tool for conservation of valuable germplasm in African buffalo. The use of aspiration, using an 18‐gauge needle and syringe, followed by slicing the ovary with a scalpel blade increased the recovery of suitable COCs (aspiration = 37.2 ± 3.02%; aspiration and slicing = 46.1 ± 3.02%), supporting the use of aspiration and slicing to optimize oocyte collection for conservation of African buffalo.