Abstract
The existence of Sulfate-Reducing Bacteria (SRB) is one of the significant reasons for the Microbially Influenced Corrosion (MIC) of jet fuel. Especially for the jet fuel stored by military, since jet fuel is stored in the tank for a long time, some oxygen-consuming bacteria such as Amorphotheca resinae and Bacillus Cohn can consume oxygen and generate organic acids at the oil-water interface of the tank bottom. This causes anaerobic SRB flourish in fuel tanks. In this study, a loop-mediated isothermal amplification (LAMP) combined with a chromatographic Lateral Flow Dipstick (LFD) assay was established to detect the SRB. Four groups of LAMP primers were designed and synthesized to target dsrB (dissimilatory sulfite reductase β-subunit) genes in SRB.LAMP-LFD can detect 121 fg/μL of SRB DNA within 35 min. The detection limit of this method is 1000 times more sensitive than the conventional PCR and shortens the detection time greatly. This method is negative for other eight common bacteria species in jet fuel, indicating that the method has high specificity. In summary, this method can be used to detect the presence of SRB in jet fuel.
Highlights
When conditions are quiescent and anaerobic, Sulfate-Reducing Bacteria (SRB) can utilize organic matter on the metal surface as a carbon source and hydrogen generated within biofilm to reduce the sulfate to hydrogen sulfide and obtain viable energy from redox reactions (Hill, 2003)
The Institute of Petroleum indicates that microbial counts in the water bottom are 109-1011 CFU/L bacteria, 107-1010 CFU/L yeasts, 106 CFU/L molds and 105 CFU/L SRB, either alone or in combination are considered to represent microbial contamination (Institute of Petroleum, 1996)
The two SRB species used in this experiment were obtained from China General Microbiological Culture Collection Center (CGMCC) and the other eight bacteria species were obtained from the China Center of Industrial Culture Collection (CICC)
Summary
When conditions are quiescent and anaerobic, SRB can utilize organic matter on the metal surface as a carbon source and hydrogen generated within biofilm to reduce the sulfate to hydrogen sulfide and obtain viable energy from redox reactions (Hill, 2003). The detection of SRB mainly relies on the traditional culture method, for example Most Probable Number (MPN) (Mccrady, 1915), which provides high precision but takes more than 15 days. Dissimilatory Sulfite Reductase (DSR) is a key enzyme in SRB dissimilatory sulfate reduction (Islamud-Din et al, 2014). The ubiquity of DSRs and their highly conserved sequences make this enzyme be suited for the detection of SRB (Ryuji et al, 2004)
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