Optimization of DNA Staining Technology for Development of Autonomous Microbe Sensor for Injection Seawater Systems

Microbial activity in the water injection system in oil and gas industry leads to an array of challenges, including biofouling, injectivity loss, reservoir plugging, and microbiologically influenced corrosion (MIC). An effective mitigation strategy requires online and real-time monitoring of microbial activity and growth in the system so that the operators can apply and adjust counter-measures quickly and properly. The previous study [1] identified DNA staining technology-with PicoGreen and SYBR Green dyes—as a very promising method for automated, online determination of microbial cell abundance in the vast Saudi Aramco injection seawater systems. This study evaluated DNA staining technology on detection limit, automation potential, and temperature stability for the construction of automated sensor prototype. DNA staining with SYBR Green dye was determined to be better suited for online and real-time monitoring of microbial activity in the Saudi Aramco seawater systems. SYBR Green staining does not require sample pre-treatment, and the fluorescence signal intensity is more stable at elevated temperatures up to 30℃. The lower detection limit of 2 × 103/ml was achieved under the optimized conditions, which is sufficient to detect microbial numbers in Saudi Aramco injection seawater. Finally, the requirements for design and construction of SYBR-based automated sensor prototype were determined.

Validation of Autonomous Microbe Sensor Prototype for Monitoring of Microorganisms in Injection Seawater Systems

Microbial growth in the water injection system is a well-known problem with severe operational and financial consequences for the petroleum industry, including microbiologically influenced corrosion (MIC), reduced injectivity, reservoir plugging, production downtime, and extensive repair costs. Monitoring of system microbiology is required in any mitigation strategy, enabling operators to apply and adjust countermeasures properly and in due time. In previous studies [1] [2], DNA staining technology with SYBR Green dye was evaluated to have a sufficient detection limit and automation potential for real-time detection of microbial activity in the Saudi Aramco injection seawater. In this study, technical requirements and design solutions were defined, and an autonomous microbe sensor (AMS) prototype was constructed, tested and optimized in the laboratory, and validated in the field for automated detection of microorganisms in the harsh Saudi Arabia desert environment and injection seawater. The AMS prototype was able to monitor and follow the general microbial status in the system, including detection of periods with increased microbial growth or decreased microbial numbers following biocide injection. The infield AMS detection limit was 105 cells/mL. The long-term field testing also identified the areas for technical improvement and optimization for further development of a more robust and better performing commercial microbial sensing device.

Applicability of Dimedone Assays for the Development of Online Aldehyde Sensor in Seawater Flooding Systems

Biocides are oilfield chemicals that are widely used to control bacterial activity throughout the oil industry. A feasibility study has been explored to develop detection techniques for biocide batch treatments, preferably on-line and in real-time, for their potential use in seawater flooding system. Several methods to measure key components of the biocide formulation were investigated and reported in previous study [1]. The enzymatic activity of an immobilized acetylcholine esterase (AChE) on the column material was successfully inhibited by some model compounds, but not by the actual biocides commonly used in Saudi Aramco seawater flooding system. In this paper, an alternative assay for biocide detection in the Saudi Aramco seawater flooding system was investigated for its applicability for the development of on-line biocide sensor. The assay was based on the detection of aldehyde functionality in the biocide mixture through measurement of a fluorescent derivative formed in the reaction of aldehyde groups and dimedone in the presence of ammonium acetate. The reaction of aldehyde groups with dimedone was demonstrated in seawater matrix, and the formed fluorescent product was successfully measured. The results showed that the dimedone-based assay was very sensitive, and relatively straightforward to perform. The ruggedness test also indicated that the assay is sensitive to minor changes of various specific conditions of the method. It is concluded that the dimedone assay is suitable for further development of a real-time biocide monitoring system to detect the presence of biocide slugs in seawater flooding system. The development of an automated on-line biocide sensor based on dimedone assay is underway.

Enhancing sensitivity of microbial fuel cell sensors for low concentration biodegradable organic matter detection:Regulation of substrate concentration,anode area and external resistance

The relatively low sensitivity is an important reason for restricting the microbial fuel cell(MFC)sensors'application in low concentration biodegradable organic matter(BOM)detection.The startup parameters,including substrate concentration,anode area and external resistance,were regulated to enhance the sensitivity of MFC sensors.The results demonstrated that both the substrate concentration and anode area were positively correlated with the sensitivity of MFC sensors,and an external resistance of 210Ωwas found to be optimal in terms of sensitivity of MFC sensors.Optimized MFC sensors had lower detection limit(1 mg/L)and higher sensitivity(Slope value of the linear regression curve was 1.02),which effectively overcome the limitation of low concentration BOM detection.The essential reason is that optimized MFC sensors had higher coulombic efficiency,which was beneficial to improve the sensitivity of MFC sensors.The main impact of the substrate concentration and anode area was to regulate the proportion between electrogens and nonelectrogens,biomass and living cells of the anode biofilm.The external resistance mainly affected the morphology structure and the proportion of living cells of the anode.This study demonstrated an effective way to improve the sensitivity of MFC sensors for low concentration BOM detection.

A self-powered biosensor based on triboelectric nanogenerator for dual-specificity bacterial detection

Pathogenic and corrosive bacteria pose a significant risk to human health or economic well-being.The specific,sensitive,and on-site detection of these bacteria is thus of paramount significance but remains challenging.Taking inspiration from immunoassays with primary and secondary antibodies,we describe here a rational design of microbial sensor(MS)under a dualspecificity recognition strategy using Pseudomonas aeruginosa(P.aeruginosa)as the detection model.In the MS,engineered aptamers are served as the primary recognition element,while polydopamine-N-acetyl-D-galactosamine(PDA-Gal NAc)nanoparticles are employed as the secondary recognition element,which will also generate and amplify changes in the output voltage signal.To achieve self-powering capability,the MS is constructed based on a triboelectric nanogenerator(TENG)with the specific aptamers immobilized on the TENG electrode surface.The as-prepared MS-TENG system exhibits good stability in output performance under external forces,and high specificity toward P.aeruginosa,with no cross-reactivity observed.A linear relationship(R^(2)=0.995)between the output voltage and P.aeruginosa concentration is established,with a limit of detection estimated at around 8.7×10^(3) CFU mL^(-1).The utilization of PDA-Gal NAc nanoparticles is found to play an important role in enhancing the specific and reliability of detection,and the underlying mechanisms are further clarified by computational simulations.In addition,the MS-TENG integrates a wireless communication module,enabling real-time monitoring of bacterial concentration on mobile devices.This work introduces a pioneering approach to designing self-powered smart microbial sensors with high specificity,using a double recognition strategy applicable to various bacteria beyond P.aeruginosa.

Laboratory-Scale Evaluation of Single Analyte Bacterial Monitoring Strategies in Water Injection Systems

Microbial activity is the cause of a variety of problems in water injection systems, e.g., microbial corrosion, plugging, and biofouling. Efficient monitoring of Saudi Aramco’s vast water injection system requires the development of online and automated technologies for monitoring microbial activities in the system. A previous system review and technology screening has identified five single-analyte strategies [1], which were evaluated in this study with a laboratory-scale setup to determine their applicability for automated determination of microbial activity in the injection water system. Four of the five single-analyte measuring principles tested in the laboratory setup were deemed less suitable for automation and/or reliable for use in the detection of microbial activity in the company injection water system. These four principles were: luminescence assay for adenosine-5’-triphosphate (ATP), detection and electrochemical measurements of H2S, determination of pH by electrochemical sensor, and measurement of oxidation-reduction potential (ORP). The strategy of staining cells with fluorescent DNA dyes, followed by quantification of fluorescence signals, was identified to hold, with proper optimization of DNA staining and fluorescence detection, a very promising potential for integration in automated, online sensors for microbial activity in the injection water system.

Real-time in situ detection and quantification of bacteria in the Arctic environment

At present,there are no methods that determine the total micro ial load on an abiotic substra tein real time.The utility of such a capability ranges from sterilization and medical diagnostics tothe search for new microorganisms in the environment and study of their ecological niches.Wereport the development of a hand held,fluorescence detection device and demonstrate its applicability to the field detection of Arctic bacteria,This technology is based on the early pioneering work of Britton Chance which elucidated the intrinsic fiuorescence of a number ofmetabolites and protein cofactors in cels,including reduced pyridine nucleotides,cytochromesand flavins.A PDA controls the device(fluorescence excitation and data collection)and processesthe multiwavelength signals to yield bacterial cell counts,including cstimates of ive cells,deadcells and endospores.Unlike existing methods for cell counting,this method requires no samplecontact or addition of reagents.The use of this technology is demonstrated with in situmea surements of two sub-glacial microbial communities at sites in Palander and colonized surfacerocks in the Bockijord Volcanic Complex during AMASE 2008(Arctic Mars Analog Svalbard Expedition),The total bacterial load on the interrogated sample surfaces ranged from<20 cells/cm to>1o9 cells/cm^(2).