Recent research indicates that the quantification of Sulfate-Reducing Bacteria (SRB) and other corrosive bacteria in biofilms is of great practical and scientific relevance. Common microbial techniques based on selective cell cultivation are of limited usefulness for both quantification and characterization of environmental populations since the used methods are time-consuming and inconvenient for field trials or express analysis. To determine the total concentration of bacteria composing the corrosive biofilms, our research team developed a new approach was developed using bioluminescent method of ATP detection. The method also was optimised to determine microorganisms number in mixed cultures. The study of kinetics of corrosive biofilm formation in artificial pilot systems of field trials were carried out. The influence of the following factors on biofilm formation was revealed: microbial content of media, presence of oil, sulfide layer on the surface and metal type. The express screening of potential biocides active in relation to corrosive biofilms and the determination of minimal concentrations of tested compounds leading to destruction of corrosive biofilms were demonstrated herein.

By using a bioluminescence ATP assay, we have determined the minimal concentrations of some biocorrosion inhibitors (Kathon, Khazar, VFIKS-82, Nitro-1, Caspii-2, and Caspii-4) suppressing most common microbial corrosion inducers: Desulfovibrio desulfuricans, Desulfovibrio vulgaris, Pseudomonas putida, Pseudomonas fluorescens, and Acidithiobacillus ferrooxidans. The cell titers determined by the bioluminescence method, including not only fissiparous cells but also their dormant living counterparts, are two-to sixfold greater than the values determined microbiologically. It is shown that the bioluminescence method can be applied to determination of cell titers in samples of oil-field waters in the presence of iron ions (up to 260 mM) and iron sulfide (to 186 mg/l) and in the absence or presence of biocidal corrosion inhibitors.

Control of oil hydrocarbon biodegradation by enumeration of oil-degrading cells was demonstrated using the bioluminescence method of intracellular ATP determination. The method was used in enumerating hydrocarbon-oxidizing bacteria in soil containing various concentrations of oil (up to 60% oil w/w) and cells (2.7×104−3.9×107 cell g−1). Investigation of the growth kinetics of Rhodococcus ruber and Pseudomonas putida in the oil-contaminated soil revealed the dependence of specific growth rate on the initial pollutant concentration. The results of the bioluminescence method correlated with levels of oil biodegradation determined gravimetrically. The apparent synergistic effect demonstrated between two cultures was verified by the bioluminescence and gravimetric methods and growth kinetics data. The same approach was used to estimate the presence of indigenous microbiota in oil-contaminated soil samples taken from the oilfields of Baku region (Azerbaijan). The total ATP level detected revealed an inverse negative relationship with oil concentration in the sample. The bioluminescence method of intracellular ATP determination could be widely used to estimate the efficacy of application of hydrocarbon-oxidizing bacteria in the decontamination processes.

By using a bioluminescence ATP assay, we have determined the minimal concentrations of some biocorrosion inhibitors (Katon, Khazar, VFIKS-82, Nitro-1, Kaspii-2, and Kaspii-4) suppressing most common microbial corrosion agents: Desulfovibrio desulfuricans, Desulfovibrio vulgaris, Pseudomonas putida, Pseudomonas fluorescens, and Acidithiobacillus ferrooxidans. The cell titers determined by the bioluminescence method, including not only dividing cells but also their dormant living counterparts, are two- to sixfold greater than the values determined microbiologically. It is shown that the bioluminescence method can be applied to determination of cell titers in samples of oil-field waters in the presence of iron ions (up to 260 mM) and iron sulfide (to 186 mg/l) and in the absence or presence of biocidal corrosion inhibitors.