Primary oil recovery is the first stage of hydrocarbon production in which a reservoir uses its natural energy to force hydrocarbon to its wellbore. Secondary oil recovery comes to play when hydrocarbons can no longer be further produced by natural means. The purpose of secondary recovery is to maintain reservoir pressure so as to displace hydrocarbons toward the wellbore. Both primary and secondary recovery processes cannot displace more than 50% of the available hydrocarbons in a reservoir. The remaining hydrocarbons are further recovered through Tertiary/Enhanced Oil Recovery techniques. According to literature, microbial enhanced oil recovery has been identified as a tertiary method used to improve the efficiency of hydrocarbon production from reservoirs. Microbial enhanced oil recovery is a feasible reservoir technology, which has not been widely used in the oil and gas industry owing to the attainment of the requisite reservoir conditions such as temperature within which microbes can thrive. Literature has shown that thermotolerant microbes can withstand optimum temperatures of 50–90°C, while deep and ultra-deep hydrocarbon reservoir temperatures are often above 100°C. This study identifies some isolated thermotolerant microbes from a sandstone reservoir that can withstand temperatures as high as 110°C via conventional methods and molecular analysis. The identified thermotolerant petroleum microbes: Bacillus amyloliquefaciens (A) and Bacillus nealsonii (B) were used to enhance oil recovery from a reservoir. The results showed that the microbial species A and B at a confined pressure of 3.0 MPa and temperature of 27°C, gave 46.4% and 48.6% oil recoveries, respectively, which is comparably higher than the value (26.9%) obtained for the water flooded samples. At temperatures of 80, 90, 100, 110 and 120°C, the oil recovery results show that the recovery factor (55.2%–64.1%) of species B were higher compared to the range (46.7–57.5%) recorded for species A. At the onset of the core flooding experiments, there was an initial increment in oil recovery factor as the temperature increased from 80 to 110°C, whereas, it remained constant within 110–120°C. This trend coincides with the drop in the thermal resistance exhibited by the microbes when exposed to such conditions. The cumulative oil production from the commercial Eclipse simulation closely matched those of the experiment results, whereas, the slight difference can be attributed to the adjustment of the simulation input parameters. The experimental results show that species B can be used to enhance oil recovery at reservoir temperature conditions above 100°C.