Enhanced Oil Recovery Using Indigenous Microbiome of High Temperature Oil Reservoirs.

Crude oil is a primary energy source used for economic expansion across the world. Secondary recovery processes employed by industries to recover oil from oil wells leave behind 70% of the oil trapped in marginal and deleted zones of reservoirs. To recover the oil from depleted zones, microbial enhanced oil recovery (MEOR) tertiary processes were introduced, which involve the production of metabolites from the indigenous microbiome. In this study, the indigenous microbiota was identified as Marinobacterium sp., Silvanigrella sp., Petrothermobacter sp., Pseudomonas sp., Bacillus sp., Nitrincola sp., Halomonas sp., Uncultured Roseovarius sp., and Phaeobacter. Further, the secondary metabolites such as volatile fatty acids (ethanol, acetone, and acetate), biomass, gases (CO2, CH4), and biosurfactants were estimated through gas chromatography and FTIR spectroscopy. Once stable microbial growth was attained in the baltch media, it was optimized through response surface methodology (RSM) to minimize the process cost. The optimized media with 9 g/L of molasses, 1.75 g/L of sodium bicarbonate, and 1.25 g/L of ammonium chloride showed a significant impact on metabolite production. Additionally, core flood studies to simulate field studies were performed that represented that TeriK-1 brought a significant increment of 18.9%, which makes it suitable for MEOR field implementation. This study is one of its kind where the indigenous thermophilic sp. was successfully established and is capable of producing the secondary metabolites that aid in the MEOR process.

Boron application improves yield of rice cultivars under high temperature stress during vegetative and reproductive stages.

It is reported that high temperatures (HT) would cause a marked decrease in world rice production. In tropical regions, high temperatures are a constraint to rice production and the most damaging effect is on spikelet sterility. Boron (B) plays a very important role in the cell wall formation, sugar translocation, and reproduction of the rice crop and could play an important role in alleviating high temperature stress. A pot culture experiment was conducted to study the effect of B application on high temperature tolerance of rice cultivars in B-deficient soil. The treatments comprised of four boron application treatments viz. control (B0), soil application of 1 kg B ha-1 (B1), soil application of 2 kg B ha-1 (B2), and foliar spray of 0.2% B (Bfs); three rice cultivars viz. Annapurna (HT stress tolerant), Naveen, and Shatabdi (both HT stress susceptible); and three temperature regimes viz. ambient (AT), HT at vegetative stage (HTV), and HT at reproductive stage (HTR). The results revealed that high temperature stress during vegetative or flowering stage reduced grain yield of rice cultivars mainly because of low pollen viability and spikelet fertility. The effects of high temperature on the spikelet fertility and grain filling varied among cultivars and the growth stages of plant when exposed to the high temperature stress. Under high temperature stress, the tolerant cultivar displays higher cell membrane stability, less accumulation of osmolytes, more antioxidant enzyme activities, and higher pollen viability and spikelet fertility than the susceptible cultivars. In the present work, soil application of boron was effective in reducing the negative effects of high temperature both at vegetative and reproductive stages. Application of B results into higher grain yield under both ambient and high temperature condition over control for all the three cultivars; however, more increase was observed for the susceptible cultivar over the tolerant one. The results suggest that the exogenous application of boron had a substantial effect on cell membrane stability, sugar mobilization, pollen viability, and spikelet fertility, hence the yield. The cultivars due to their variation in the tolerance level for high temperature stress behaved differently, and at high temperature stress, more response of the application of boron was seen in susceptible cultivars.

Gut microbial diversity in health and disease: experience of healthy Indian subjects, and colon carcinoma and inflammatory bowel disease patients.

Background: The intestinal microbiota, through complex interactions with the gut mucosa, play a key role in the pathogenesis of colon carcinoma and inflammatory bowel disease (IBD). The disease condition and dietary habits both influence gut microbial diversity. Objective: The aim of this study was to assess the gut microbial profile of healthy subjects and patients with colon carcinoma and IBD. Healthy subjects included 'Indian vegetarians/lactovegetarians', who eat plant produce, milk and milk products, and 'Indian non-vegetarians', who eat plant produce, milk and milk products, certain meats and fish, and the eggs of certain birds and fish. 'Indian vegetarians' are different from 'vegans', who do not eat any foods derived wholly or partly from animals, including milk products. Design: Stool samples were collected from healthy Indian vegetarians/lactovegetarians and non-vegetarians, and colon cancer and IBD patients. Clonal libraries of 16S ribosomal DNA (rDNA) of bacteria were created from each sample. Clones were sequenced from one representative sample of each group. Approximately 500 white colonies were picked at random from each sample and 100 colonies were sequenced after amplified rDNA restriction analysis. Results: The dominant phylum from the healthy vegetarian was Firmicutes (34%), followed by Bacteroidetes (15%). The balance was reversed in the healthy non-vegetarian (Bacteroidetes 84%, Firmicutes 4%; ratio 21:1). The colon cancer and IBD patients had higher percentages of Bacteroidetes (55% in both) than Firmicutes (26% and 12%, respectively) but lower Bacteroidetes:Firmicutes ratios (3.8:1 and 2.4:1, respectively) than the healthy non-vegetarian. Bacterial phyla of Verrucomicrobiota and Actinobacteria were detected in 23% and 5% of IBD and colon patients, respectively. Conclusions: Ribosomal Database Project profiling of gut flora in this study population showed remarkable differences, with unique diversity attributed to different diets and disease conditions.

Electrochemical removal of sulfate from petroleum produced water.

Petroleum produced water (PPW) is a waste-stream that entails huge cost on the petroleum industry. Along with other suspended and dissolved solids, it contains sulfate, which is a major hurdle for its alternative use intended toward enhanced oil recovery. This study proposes a two-step process for sulfate removal from PPW. A synthetic PPW was designed for the study using response surface methodology. During the first step, sulfate present in PPW was reduced to sulfide by anaerobic fermentation with 80% efficiency. In the second step, more than 70% of the accumulated sulfide was electrochemically oxidized. This integrated approach successfully removed sulfate from the synthetic wastewater indicating its applicability in the treatment of PPW and its subsequent applications in other oil field operations.

Bioleaching of nickel from spent petroleum catalyst using Acidithiobacillus thiooxidans DSM- 11478.

The present work deals with optimization of culture conditions and process parameters for bioleaching of spent petroleum catalyst collected from a petroleum refinery. The efficacy of Ni bioleaching from spent petroleum catalyst was determined using pure culture of Acidithiobacillus thiooxidans DSM- 11478. The culture conditions of pH, temperature and headspace volume to media volume ratio were optimized. EDX analysis was done to confirm the presence of Ni in the spent catalyst after roasting it to decoke its surface. The optimum temperature for A. thiooxidans DSM-11478 growth was found to be 32 degrees C. The enhanced recovery of nickel at very low pH was attributed to the higher acidic strength of sulfuric acid produced in the culture medium by the bacterium. During the bioleaching process, 89% of the Ni present in the catalyst waste could be successfully recovered in optimized conditions. This environment friendly bioleaching process proved efficient than the chemical method. Taking leads from the lab scale results, bioleaching in larger volumes (1, 5 and 10 L) was also performed to provide guidelines for taking up this technology for in situ industrial waste management.

Application of bioflocculating property of Pseudomonas aeruginosa strain IASST201 in treatment of oil-field formation water.

A bioflocculating activity of 89.8% was depicted by an activated sludge-borne bacteria Pseudomonas aeruginosa strain IASST201 with a yield of bioflocculant of 2.68 g L(-1) obtained from production media broth after optimization of different parameters. The highest bioflocculation efficiency was found at the pre-stationary phase of the bacterial growth period in the production media broth at 96th hour examined from a growth-flocculation kinetics study. 85.67% of bioflocculation was observed in oil-field formation water, with a separation of 68.7% of aliphatic hydrocarbon contents of the formation water after the application of the bacterial bioflocculant by entrapment mechanism with formation of flocs which was analyzed and examined comparatively through gas-chromatography. Extensive removal of heavy metal contents of the oil-field formation water due to bioflocculation was estimated by Atomic Absorption Spectrophotometer (AAS). The SEM and AFM studies declare the extracellular polymeric nature of the bioflocculant produced by this bacterium clumped within bacterial biofilm supported with FTIR study of the extracted bioflocculant.

Laboratory investigation and core flood demonstration of enhanced biogenic methane generation from lignite.

Over the last several decades, coalbed methane (CBM) has emerged as an important energy source in developing nations like India as well as worldwide and is expected to play a significant role in the energy portfolio of the future. The current scenario of rapid exhaustion of fossil fuels is leading to the need to explore alternative and efficient fuel resources. The present study demonstrates enhanced methane production per gram of lignite (lowest-rank coal). Optimization of the bioconversion of lignite to methane revealed 55°C temperature and 1.5 g/L NaCl concentration as ambient conditions for the process. A scale-up study in the optimized condition showed 2,800 mM methane production per 25 g of lignite in anaerobic conditions. Further, Fourier transform Infrared (FTIR) and Gas Chromatography Mass Spectrometry (GCMS) analysis showed bioconversion of lignite into simpler intermediate substrates required for methane production. The results highlighted that the bacterial action first converts lignite into volatile fatty acids, which subsequently get converted into methane. Further, the exploration of indigenous microbial consortia in Tharad well (THAA) mainly comprises the order Methanosarcinales and Methanomicrobiales. The pathogenicity of the microbial consortium THAA was declared safe for use in mice via the oral route by The Energy and Resources Institute (TERI), India. The study demonstrated the development of indigenous consortia (TERI THAA), which can potentially enhance methane production from the lowest coal grade under extreme conditions in Indian coal beds.

Isolation of a lead tolerant novel bacterial species, Achromobacter sp. TL-3: assessment of bioflocculant activity.

Lead is one of the four heavy metals that has a profound damaging effects on human health. In the recent past there has been an increasing global concern for development of sustainable bioremediation technologies for detoxification of lead contaminant. Present investigation highlights for lead biosorption by a newly isolated novel bacterial species; Achromobacter sp. TL-3 strain, isolated from activated sludge samples contaminated with heavy metals (collected from oil refinery, Assam, North-East India). For isolation of lead tolerant bacteria, sludge samples were enriched into Luria Broth medium supplemented separately with a range of lead nitrate; 250, 500, 750, 1000, 1250 and 1500 ppm respectively. The bacterial consortium that could tolerate 1500 ppm of lead nitrate was selected further for purification of lead tolerant bacterial isolates. Purified lead tolerant bacterial isolates were then eventually inoculated into production medium supplemented with ethanol and glycerol as carbon and energy source to investigate for bioflocculant production. Bioflocculant production was estimated by monitoring the potential of lead tolerant bacterial isolate to flocculate Kaolin clay in presence of 1% CaCl2. Compared to other isolates, TL-3 isolate demonstrated for maximum bioflocculant activity of 95% and thus was identified based on 16S rRNA gene sequence analysis. TL3 isolate revealed maximum homology (98%) with Achromobacter sp. and thus designated as Achromobacter sp. TL-3. Bioflocculant activity of TL-3 isolate was correlated with the change in pH and growth. Achromobacter sp. TL-3 has significant potential for lead biosorption and can be effectively employed for detoxification of lead contaminated waste effluents/waste waters.

Biosurfactant: an emerging tool for the petroleum industries.

The petroleum sector is essential to supplying the world's energy demand, but it also involves numerous environmental problems, such as soil pollution and oil spills. The review explores biosurfactants' potential as a new tool for the petroleum sector. Comparing biosurfactants to their chemical equivalents reveals several advantages. They are ecologically sustainable solutions since they are renewable, nontoxic, and biodegradable. Biosurfactants are used in a variety of ways in the petroleum sector. They can improve the mobilization and extraction of trapped hydrocarbons during oil recovery procedures. By encouraging the dispersion and solubilization of hydrocarbons, biosurfactants also assist in the cleanup of oil spills and polluted locations by accelerating their breakdown by local microorganisms. The review gives insights into alternative methods for the petroleum industry that are more viable and cost-effective.

Culture-independent assessment of the indigenous microbial diversity of Raniganj coal bed methane block, Durgapur.

It is widely acknowledged that conventional mining and extraction techniques have left many parts of the world with depleting coal reserves. A sustainable method for improving the recovery of natural gas from coalbeds involves enhancing the production of biogenic methane in coal mines. By taking a culture-independent approach, the diversity of the microbial community present in the formation water of an Indian reservoir was examined using 16S rRNA gene amplification in order to study the potential of microbial-enhanced coal bed methane (CBM) production from the deep thermogenic wells at a depth of 800-1200 m. Physicochemical characterization of formation water and coal samples was performed with the aim of understanding the in situ reservoir conditions that are most favorable for microbial CBM production. Microbial community analysis of formation water showed that bacteria were more abundant than archaea. Proteobacteria, Firmicutes, and Bacteroidetes were found as the most prevalent phyla in all the samples. These phyla play a crucial role in providing substrate for the process of methanogenesis by performing fermentative, hydrolytic, and syntrophic functions. Considerable variation in the abundance of microbial genera was observed amongst the selected CBM wells, potentially due to variable local geochemical conditions within the reservoir. The results of our study provide insights into the impact of geochemical factors on microbial distribution within the reservoir. Further, the study demonstrates lab-scale enhancement in methane production through nutrient amendment. It also focuses on understanding the microbial diversity of the Raniganj coalbed methane block using amplicon sequencing and further recognizing the potential of biogenic methane enhancement through microbial stimulation. The findings of the study will help as a reference for better strategization and implementation of on-site microbial stimulation for enhanced biogenic methane production in the future.

Nutrient optimization for indigenous microbial consortia of a Bhagyam oil field: MEOR studies.

The microbial enhanced oil recovery (MEOR) method is an eco-friendly and economical alternative technology. The technology involves a variety of uncertainties, and its success depends on controlling microbial growth and metabolism. This study is one of a kind that showed successful tertiary recovery of crude oil through indigenous microbial consortia. In this study, a medium was optimized to allow ideal microbial growth under reservoir conditions through RSM. Once the nutrient recipe was optimized, the microbial metabolites were estimated through gas chromatography. The maximum amount of methane gas (0.468 mM) was produced in the TERIW174 sample. The sequencing data set showed the presence of Methanothermobacter sp. and Petrotoga sp. In addition, these established consortia were analyzed for their toxicity, and they appeared to be safe for the environment. Furthermore, a core flood study showed efficient recovery that was ~25 and 34% in TERIW70 and TERIW174 samples, respectively. Thus, both the isolated consortia appeared to be suitable for the field trials.

Exhaust particle number and composition for diesel and gasoline passenger cars under transient driving conditions: Real-world emissions down to 1.5 nm.

Recent recommendations given by WHO include systematic measurements of ambient particle number concentration and black carbon (BC) concentrations. In India and several other highly polluted areas, the air quality problems are severe and the need for air quality related information is urgent. This study focuses on particle number emissions and BC emissions of passenger cars that are technologically relevant from an Indian perspective. Particle number and BC were investigated under real-world conditions for driving cycles typical for Indian urban environments. Two mobile laboratories and advanced aerosol and trace gas instrumentation were utilized. Our study shows that passenger cars without exhaust particle filtration can emit in real-world conditions large number of particles, and especially at deceleration a significant fraction of particle number can be even in 1.5-10 nm particle sizes. The mass concentration of exhaust plume particles was dominated by BC that was emitted especially at acceleration conditions. However, exhaust particles contained also organic compounds, indicating the roles of engine oil and fuel in exhaust particle formation. In general, our study was motivated by serious Indian air quality problems, by the recognized lack of emission information related to Indian traffic, and by the recent WHO air quality guidance; our results emphasize the importance of monitoring particle number concentrations and BC also in Indian urban areas and especially in traffic environments where people can be significantly exposed to fresh exhaust emissions.

Biodegradation of asphalt by Garciaella petrolearia TERIG02 for viscosity reduction of heavy oil.

Petroleum hydrocarbon is an important energy resource, but it is difficult to exploit due to the presence of dominated heavy constituents such as asphaltenes. In this study, viscosity reduction of Jodhpur heavy oil (2,637 cP at 50°C) has been carried out by the biodegradation of asphalt using a bacterial strain TERIG02. TERIG02 was isolated from sea buried oil pipeline known as Mumbai Uran trunk line (MUT) located on western coast of India and identified as Garciaella petrolearia by 16S rRNA full gene sequencing. TERIG02 showed 42% viscosity reduction when asphalt along with molasses was used as a sole carbon source compared to only asphalt (37%). The viscosity reduction by asphaltene degradation has been structurally characterized by Fourier transform infrared spectroscopy (FTIR). This strain also shows an additional preference to degrade toxic asphalt and aromatics compounds first unlike the other known strains. All these characteristics makes TERIG02 a potential candidate for enhanced oil recovery and a solution to degrading toxic aromatic compounds.

Biodegradation of oil spill by petroleum refineries using consortia of novel bacterial strains.

Feasibility study carried out at the site prior to the full scale study showed that the introduced bacterial consortium effectively adapted to the local environment of the soil at bioremediation site. The soil samples were collected from the contaminated fields after treatment with bacterial consortium at different time intervals and analyzed by gas chromatography after extraction with hexane and toluene. At time zero (just before initiation of bioremediation), the concentration of total petroleum hydrocarbons in the soil (25-cm horizon) of plot A, B, C and D was 30.90 %, 18.80 %, 25.90 % and 29.90 % respectively, after 360 days of treatment with microbial consortia was reduced to 0.97 %, 1.0 %, 1.0 %, and 1.1 % respectively. Whereas, only 5 % degradation was observed in the control plot after 365 days (microbial consortium not applied).

A metagenomic assessment of gut microbiota in Indian colon cancer patients.

Background: Gut microbiota plays an important role in the development of different diseases including colorectal cancer. The geography, lifestyle, and dietary habits of Indians are different from Western world, thus microbiome studies of Western population could not be extrapolated to their Indian counterparts. Method: Therefore, we have conducted a study on gut microbiota in Indian healthy subjects and patients of colon cancer using 16S ribosomal RNA Amplicon sequencing. Operational taxonomic units were calculated for different bacterial taxon including phylum, class, order, family, and genus level. Results: Observed results indicated a considerable difference in the bacterial diversity in both the groups. Phylum Firmicutes was significantly dominated in both the groups followed by Bacteroidetes, Actinobacteria, and Proteobacteria which clearly indicates the dominance of phylum Firmicutes in Indian population. Phylum Firmicutes and Actinobacteria were significantly abundant in the healthy group while phylum Bacteroidetes in the colon cancer group. Bacterial genera Megamonas, Megasphaera, Mitsuokella, and Streptococcus were significantly abundant in the healthy group and Veillonella, Prevotella, and Eubacterium in the colon cancer group. Bacterial genus Bradyrhizobium was present in the healthy group and Alistipes, Coprococcus, Dorea, and Rhodococcus were present in the colon cancer group but absent in the healthy group. Conclusion: There was a considerable difference in bacterial diversity in both the study groups indicating dysbiosis in the colon cancer group.

Synthesis, characterization, method development, and validation of nor-ethylmorphine hydrochloride reference material using established analytical techniques for dope control analysis.

Ethylmorphine is permitted internationally for therapeutic purposes where morphine is not indicated across the globe. Nor-ethylmorphine a major metabolite of ethylmorphine. To differentiate the intake of morphine from ethylmorphine, nor-ethylmorphine stable reference material is desirable. There is no available commercial source and no data for reference material context for this substance. Therefore, nor-ethylmorphine HCl was synthesized and characterized, and purity and potency were assessed using nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HRMS), Fourier transform infrared spectroscopy (FT-IR), thermogravimetry (TGA), and high-performance liquid chromatography (HPLC). Purity and potency were found to be 98.29% and 96.40%, respectively, providing a fit for purpose reference material for doping control analysis in sports.

Oceanotoga teriensis gen. nov., sp. nov., a thermophilic bacterium isolated from offshore oil-producing wells.

A novel, moderately thermophilic, chemo-organotrophic bacterium was isolated from formation fluid samples from an offshore oil-production well head at Bombay High (Western India). Cells were rod-shaped with a sheath-like outer structure ('toga'); the cells appeared singly, in pairs or in short chains. Cells grew at 25-70 °C (optimum 55-58 °C), pH 5.5-9.0 (optimum pH 7.3-7.8) and 0-12  % (w/v) NaCl (optimum 4.0-4.5  %). The isolate was able to grow on various carbohydrates or complex proteinaceous substances. The isolate reduced thiosulfate and elemental sulfur. The major end products of glucose fermentation were acetate, H2 and CO2. The DNA G+C content of the genomic DNA was 26.8 mol%. Phylogenetic analysis of the 16S rRNA gene placed the strain within the order Thermotogales in the bacterial domain. On the basis of 16S rRNA gene sequence comparisons and in combination with morphological and physiological characteristics, the isolate represents a novel species of new genus, for which the name Oceanotoga teriensis gen. nov., sp. nov. is proposed. The type strain of the type species is OCT74(T) (=JCM 15580(T)=LMG 24865(T)).

Efficacy of natural biocide on control of microbial induced corrosion in oil pipelines mediated by Desulfovibrio vulgaris and Desulfovibrio gigas.

We compared the efficacy of a natural biocide with four chemical tetrakishydroxymethyl phosphonium sulfonate, benzyl trimethyl ammonium chloride, and formaldehyde, glutaraldehyde, to control microbial induced corrosion in oil pipelines. The efficacy of biocides were monitored against Desulfovibrio vulgaris and Desulfovibrio gigas in experimental pipes by measuring cell counts, H2S production, Fe(II) production, production of extracellular polymeric substances and structure of biofilm. The treatment with cow urine had minimum planktonic cell counts of 3 x 10(2) CFU/mL as well as biofilm cell counts of 9 x 10(1) CFU/mL as compared with tetrakishydroxyl methyl phosphonium sulfonate, benzyl trimethyl ammonium chloride, formaldehyde and glutaraldehyde. Sulfide production was the lowest with cow urine (0.08 mmol/L), followed by tetrakishydroxymethyl phosphonium sulfonate 0.72 mmol/L. On day 90 of treatment, Fe(II) production was also found to be the lowest with cow urine. The scanning electron microscopic studies indicated that the biofilm bacteria were killed by cow urine. These results demonstrate the cow urine mediated control of microbially induced corrosion, and this is indicative of its potential as a viable substitute of toxic biocides. To the best of our knowledge, this seems to be the first report which screens possible biocidal activity by cow urine as compared to the most common biocides which oil industry is currently using.

Identification and probiotic potential of lactic acid bacteria from camel milk.

In the present study, a total of 80 presumed lactic acid bacteria (LAB) were isolated from camel milk. Selected LAB were identified as Lactococcus lactis (cam 12), Enterococcus lactis (cam 14) and Lactobacillus plantarum (cam 15) and their potential were tested by tolerance & de-conjugation of bile salts, antimicrobial activity, surface hydrophobicity and adhesion potential) along with this of probiotics were evaluated for curd formation and assessed for sensory properties and syneresis. Selected LABs showed antimicrobial activity against wide range of pathogenic bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus cereus and Escherchiaia. coli). LAB (cam 12, cam 14 and cam15) were highly sceptible to chloramphenicol, vancomycin, and tetracyclin. In vitro adhesion studies with Caco-2 cells demonstrated strong adhesion activity with hydrophobicity (99%) was observed. Acute oral toxicity of E. lactis and L. plantarum showed non-toxic, non-virulent and safe for industrial application. The study provides potential LAB which may act as a substitute of functional food, synthetic feed and industrial curd formulation with in the shortest span (240 min at 28-32 °C).