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Autoclaving of crude oil is often used to evaluate the hydrocarbon-degrading abilities of bacteria. This may be potentially useful for bioaugmentation and microbial enhanced oil recovery (MEOR). However, it is not entirely clear if “endogenous” bacteria (e.g., spores) in/on crude oil survive the autoclaving process, or influence subsequent evaluation of the hydrocarbon-degradation abilities of the “exogenous” bacterial strains. To test this, we inoculated autoclaved crude oil medium with six exogenous bacterial strains (three  Dietzia  strains, two  Acinetobacter  strains, and one  Pseudomonas  strain). The survival of the spore-forming  Bacillus  and  Paenibacillus  and the non-spore-forming mesophilic  Pseudomonas, Dietzia, Alcaligenes , and  Microbacterium  was detected using a 16S rRNA gene clone library and terminal restriction fragment length polymorphism (T-RFLP) analysis. However, neither bacteria nor bacterial activity was detected in three controls consisting of non-inoculated autoclaved crude oil medium. These results suggest that detection of endogenous bacteria was stimulated by the six inoculated strains. In addition, inoculation with  Acinetobacter  spp. stimulated detection of  Bacillus , while inoculation with  Dietzia  spp. and  Pseudomonas  sp. stimulated the detection of more  Pseudomonas . In contrast, similar exogenous bacteria stimulated similar endogenous bacteria at the genus level. Based on these results, special emphasis should be applied to evaluate the influence of bacteria capable of surviving autoclaving on the hydrocarbon-degrading abilities of exogenous bacteria, in particular, with regard to bioaugmentation and MEOR. Bioaugmentation and MEOR technologies could then be developed to more accurately direct the growth of specific endogenous bacteria that may then improve the efficiency of treatment or recovery of crude oil.

Bacteria in Crude Oil Survived Autoclaving and Stimulated Differentially by Exogenous Bacteria