Synthesis and characterization of iron oxide nanoparticles from Lawsonia inermis and its effect on the biodegradation of crude oil hydrocarbon.

Crude oil hydrocarbons are considered major environmental pollutants and pose a significant threat to the environment and humans due to having severe carcinogenic and mutagenic effects. Bioremediation is one of the practical and promising technology that can be applied to treat the hydrocarbon-polluted environment. In this present study, rhamnolipid biosurfactant (BS) produced by Pseudomonas aeruginosa PP4 and green synthesized iron nanoparticles (G-FeNPs) from Lawsonia inermis was used to evaluate the biodegradation efficiency (BE) of crude oil. The surface analysis of G-FeNPs was carried out by using FESEM and HRTEM to confirm the size and shape. Further, the average size of the G-FeNPs was observed around 10 nm by HRTEM analysis. The XRD and Raman spectra strongly confirm the presence of iron nanoparticles with their respective peaks. The BE (%) of mixed degradation system-V (PP4+BS+G-FeNPs) was obtained about 82%. FTIR spectrum confirms the presence of major functional constituents (C=O, -CH3, C-O, and OH) in the residual oil content. Overall, this study illustrates that integrated nano-based bioremediation could be an efficient approach for hydrocarbon-polluted environments. This study is the first attempt to evaluate the G-FeNPs with rhamnolipid biosurfactant on the biodegradation of crude oil.

Biodegradation of pyrene by bacterial consortia: Impact of natural surfactants and iron oxide nanoparticles.

Polycyclic aromatic hydrocarbons (PAHs) are potentially hazardous compounds that could cause a severe impact on many ecosystems. They are very challenging to remove using conventional methods due to their hydrophobic nature. However, this issue can be resolved by utilizing surface-active molecules to increase their bioavailability. In this study, pyrene was chosen as the PAH compound to explore its degradability by the effect of individual bacterial strains (Pseudomonas stutzeri NA3 and Acinetobacter baumannii MN3) and mixed consortia (MC) along with natural surfactant derived from Sapindus mukorossi and iron oxide nanoparticles (NPs). Additionally, fatty acids esters, dipeptides, and sugar derivative groups were identified as potent bioactive components of natural surfactants. Various techniques, such as XRD, VSM, TEM, and FE-SEM with EDX, were utilized to characterize the pristine and Fenton-treated iron oxide NPs. The analytical results confirmed that the Fe3O4 crystal phase and spherical-shaped NPs exhibited excellent magnetic properties. The impact of natural surfactants and iron oxide NPs has significantly contributed to the biodegradation process, resulting in a prominent decrease in chemical oxygen demand (COD) levels. Gas chromatography-mass spectrometry (GC-MS) analysis showed that biodegradation systems produced primary hydrocarbon intermediates, which underwent oxidative degradation through Fenton treatment. Interestingly, synthesized iron oxide NPs effectively produced hydroxyl radical (•OH) during the Fenton reaction, which was confirmed by electron paramagnetic resonance (EPR) spectra, and the pristine iron oxide NPs underwent a material transformation observed. The study demonstrated an integrated approach for biodegradation and the Fenton reaction process to enhance the pyrene degradation efficiency (90%) compared to other systems. Using natural surfactants and iron oxide NPs in aquatic environments serves as a crucial platform at the interface of microorganisms and contaminated oil products. This interaction offers a promising solution for PAHs bioremediation.

Integrated approach of nano assisted biodegradation of anthracene by Pseudomonas aeruginosa and iron oxide nanoparticles.

Poly aromatic hydrocarbons (PAHs) are considered as hazardous compounds which causes serious threat to the environment dua to their more carcinogenic and mutagenic impacts. In this study, Pseudomonas aeruginosa PP4 strain and synthesized iron nanoparticles were used to evaluate the biodegradation efficiency (BE %) of residual anthracene. The BE (%) of mixed degradation system (Anthracene + PP4+ FeNPs) was obtained about 67 %. The FTIR spectra result revealed the presence of functional groups (C-H, -CH3, CC, =C-H) in the residual anthracene. The FESEM and TEM techniques were used to determine the surface analysis of the synthesized FeNPs and the average size was observed by TEM around 5-50 nm. The crystalline nature of the synthesized iron nanoparticles was confirmed by the observed different respective peaks of XRD pattern. The various functional constituents (OH, C-H, amide I, CH3) were identified in the synthesized iron nanoparticles by FTIR spectrum. In conclusion, this integrated nano-bioremediation approach could be an promising and effective way for many environmental fields like cleanup of hydrocarbon rich environment.

Impact of biosurfactant produced by Bacillus spp. on biodegradation efficiency of crude oil and anthracene.

Biosurfactants are surface active molecules generated by various microorganisms, including bacteria, actinobacteria, algae, and fungi. In this study, bacterial strains are isolated from soil contaminated with used motor oil and examined for potential biosurfactant production. A minimum salt medium (MSM), with crude oil as the only carbon source, is used to isolate potential biosurfactant-producing bacterial strains. About 23 strains are isolated, and all are subjected to the primary screening methods for biosurfactant production. Based on the emulsification index, oil displacement, and drop collapse screening methods, two isolates with potential biosurfactant-producing ability are selected for further studies. The synthesis of biosurfactants, crude oil and anthracene biodegradation is carried out with strains DTS1 and DTS2. Both strains show significant outcomes in crude oil degradation. In addition, both strains can utilize anthracene as the sole carbon source. During the degradation course, changes in the growth conditions are continuously monitored by measuring turbidity and pH. In this degradation study, the biosurfactant production aptitude of the isolated strains plays an essential role in increasing the bioavailability of hydrophobic hydrocarbons. These strains are identified down to the molecular level by employing 16S rRNA gene sequencing, and the acquired sequences are submitted to get the accession numbers. These prospective strains can be utilized to remediate hydrocarbon-contaminated environments.

Impact of biosurfactant and iron nanoparticles on biodegradation of polyaromatic hydrocarbons (PAHs).

Polycyclic aromatic hydrocarbons (PAHs) are hazardous toxic contaminants and considered as primary pollutants due to their persistent nature and most of them are carcinogenic and mutagenic. The key challenge in PAHs degradation is their hydrophobic nature, which makes them one of the most complex materials and inaccessible by a broad range of microorganisms. This bioavailability can be increased by using a biosurfactant. In the present study mixed PAHs were degraded using the biosurfactant producing bacterial strains. In addition, iron nanoparticles were synthesized and the impact of iron nanoparticles on the growth of the mixed bacterial strains (Pseudomonas stutzeri NA3 and Acinetobacter baumannii MN3) was optimized. The mixed PAHs (anthracene, pyrene, and benzo(a)pyrene) degradation was enhanced by addition of biosurfactant (produced by Bacillus subtilis A1) and iron nanoparticles, resulting in 85% of degradation efficiency. The addition of the biosurfactant increased the bioavailability of the PAHs in the aqueous environment, which might help bacterial cells for the initial settlement and development. The addition of iron nanoparticles increased both bacterial biomass and PAHs adsorption over their surface. These overall interactions assisted in the utilization of PAHs by the mixed bacterial consortia. This study illustrates that this integrated approach can be elaborated for the removal of the complex PAHs pollutants from soil and aqueous environments.

Characterization of bacterial community in oil-contaminated soil and its biodegradation efficiency of high molecular weight (>C40) hydrocarbon.

In this study, two biosurfactant producing Pseudomonas aeruginosa sp. were isolated from motor oil contaminated soil for crude oil, alkane and PAH degradation studies. Metagenomics analysis identified as proteobacteria phyla was the dominant. Isolated two bacterial species were well grown in mineral salt medium with 1% of crude oil, alkanes (dotriacontane and tetratetracontane) and PAH (pyrene, benzopyrene and anthracene) as sole carbon sources. Total biodegradation efficiency (BE) of strains PP3 and PP4 in Crude oil degradation evaluated by the analysis of gas chromatography and mass spectrometry was 50% and 86% respectively. BE of PP3, PP4 and mixed consortium in alkane biodegradation were 46%, 47% and 36%, respectively. BE of PP3, PP4 and mixed consortium in PAH biodegradation were 22%, 48% and 35%, respectively. Based on the results revealed that strain pp4 was more efficient bacteria to degrade the crude oil, alkane and PAH than pp3. This was due to the higher production of biosurfactant by PP4 than PP3 and also confirmed in the test of emulsification index (E24). FTIR results showed that the produced biosurfactant could partially solubilize the crude oil hydrocarbons, alkanes and PAH and confirmed as glycolipid (rhamnolipid) in nature. Thus, the obtained results from the GCMS showed that all hydrocarbons were utilized by bacteria as carbon source for biosurfactant production and utilize the high molecular weight hydrocarbons. Based on the present study we can suggest that identified potential biosurfactant producing bacteria are used for biodegradation of high molecular weight hydrocarbon (>C40).

Characterization of crude oil degrading bacterial communities and their impact on biofilm formation.

In the present study, produced water sample collected from the Indian crude oil reservoir is used to enrich the bacterial communities. The impact of these enriched bacterial communities on the biodegradation of crude oil, biofilm formation, and biocorrosion process are elucidated. A crude oil degradation study is carried out with the minimal salt medium and 94% of crude oil was utilized by enriched bacterial communities. During the crude oil degradation many enzymes including alkane hydroxylase, alcohol dehydrogenase, and lipase are playing a key role in the biodegradation processes. The role of enriched bacterial biofilm on biocorrosion reactions are monitored by weight loss studies and electrochemical analysis. Weight loss study revealed that the biotic system has vigorous corrosion attacks compared to the abiotic system. Both AC-Impedance and Tafel analysis confirmed that the nature of the corrosion reaction take place in the biotic system. Very less charge transfer resistance and higher corrosion current are observed in the biotic system than in the abiotic system. Scanning electron microscope confirms that the dense biofilm formation favoured the pitting type of corrosion. X-ray diffraction analysis confirms that the metal oxides formed in the corrosion systems (biotic). From the metagenomic analysis of the V3-V4 region revealed that presence of diverse bacterial communities in the biofilm, and most of them are uncultured/unknown. Among the known genus, Bacillus, Halomonas, etc are dominant in the enriched bacterial biofilm sample. From this study, we conclude that the uncultured bacterial strains are found to be playing a key role in the pitting type of corrosion and they can utilize crude oil hydrocarbons, which make them succeeded in extreme oil reservoir environments.

Evaluation of Syzygium aromaticum aqueous extract as an eco-friendly inhibitor for microbiologically influenced corrosion of carbon steel in oil reservoir environment.

In the present investigation, biocorrosion inhibition efficiency of Syzygium aromaticum (clove) aqueous extract on carbon steel in presence of four corrosion causing bacterial strains (Bacillus subtilis, Streptomyces parvus, Pseudomonas stutzeri, and Acinetobacter baumannii) was explored. Weight loss, potentiodynamic polarization, and AC impedance studies were carried out with and without bacterial strains and clove extract. The results obtained from weight loss and AC impedance studies indicate that these corrosion causing bacterial strains accelerated the biocorrosion reaction and biofilm playing a key role in this process. However, the addition of clove extract into the corrosive medium decreased the corrosion current and increased the solution and charge transfer resistance. The significant inhibition efficiency of about 87% was archived in the mixed consortia system with clove extract. The bioactive compounds were playing an important role in the antibacterial activity of the clove extract. It was revealed that clove extract has both biocidal and corrosion inhibition properties.

Bio-electrokinetic remediation of crude oil contaminated soil enhanced by bacterial biosurfactant.

The present study evaluating the coupling between bioremediation (BIO) and electrokinetic (EK) remediation of crude oil hydrocarbon by using bio-electrokinetic (BIO-EK) technique. The application of bacterial biosurfactant (BS) may increase the remediation efficiency by increasing the solubility of organic materials. In this work, the potential biosurfactant producing marine bacteria were isolated and identified by 16S rDNA analysis namely Bacillus subtilis AS2, Bacillus licheniformis AS3 and Bacillus velezensis AS4. Biodegradation efficiency of crude oil was found as 88%, 92% and 97% for strain AS2, AS3 and AS4 respectively, with the optimum temperature of 37 °C and pH 7. FTIR confirm the BS belongs to lipopeptide in nature. GCMS reveals that three isolates degraded the lower to higher molecular weight of the crude oil (C8 to C28) effectively. Results showed that use of BS in electokinetic remediation enhance the biodegradation rate of crude oil contaminated soil about 92% than EK (60%) in 2 days operation. BS enhances the solubilization of hydrocarbon and it leads to the faster electromigration of hydrocarbon to the anodic compartment, which was confirmed by the presence of higher total organic content than the EK. This study proven that the BIO-EK combined with BS can be used to enhance in situ bioremediation of petroleum contaminated soils.

Bacterial community analysis of biofilm on API 5LX carbon steel in an oil reservoir environment.

This study aimed to characterize the biofilm microbial community that causes corrosion of API 5LX carbon steel. API 5LX carbon steel coupons were incubated with raw produced water collected from two oil reservoir stations or filter-sterilized produced water. Biofilm 16S rRNA amplicon sequencing revealed that the bacterial community present in the biofilm was dominated by Proteobacteria, including Marinobacter hydrocarbonoclaustics and Marinobacter alkaliphilus. Electrochemical analysis such as impedance and polarization results indicated that Proteobacteria biofilm accelerated corrosion by ~ twofold (2.1 ± 0.61 mm/years) or ~ fourfold (~ 3.7 ± 0.42 mm/years) when compared to the control treatment (0.95 ± 0.1 mm/years). Scanning electron and atomic force microscopy revealed the presence of a thick biofilm and pitting corrosion. X-ray diffraction revealed higher amounts of the corrosion products Fe2O3, γ-FeOOH, and α-FeOOH, and confirmed that the microbial biofilm strongly oxidized the iron and contributed to the acceleration of corrosion of carbon metal API 5LX.

Role of thermophilic bacteria (Bacillus and Geobacillus) on crude oil degradation and biocorrosion in oil reservoir environment.

Thermophilic bacterial communities generate thick biofilm on carbon steel API 5LX and produce extracellular metabolic products to accelerate the corrosion process in oil reservoirs. In the present study, nine thermophilic biocorrosive bacterial strains belonging to Bacillus and Geobacillus were isolated from the crude oil and produced water sample, and identified using 16S rRNA gene sequencing. The biodegradation efficiency of hydrocarbons was found to be high in the presence of bacterial isolates MN6 (82%), IR4 (94%) and IR2 (87%). During the biodegradation process, induction of the catabolic enzymes such as alkane hydroxylase, alcohol dehydrogenase and lipase were also examined in these isolates. Among them, the highest activity of alkane hydroxylase (130 µmol mg-1 protein) in IR4, alcohol dehydrogenase (70 µmol mg-1 protein) in IR2, and higher lipase activity in IR4 (60 µmol mg-1 protein) was observed. Electrochemical impedance spectroscopy and X-ray diffraction data showed that these isolates oxidize iron into ferrous/ferric oxides as the corrosion products on the carbon steel surface, whilst the crude oil hydrocarbon served as a sole carbon source for bacterial growth and development in such extreme environments.

Bioengineered silver nanoparticles as potent anti-corrosive inhibitor for mild steel in cooling towers.

Silver nanoparticle-aided enhancement in the anti-corrosion potential and stability of plant extract as ecologically benign alternative for microbially induced corrosion treatment is demonstrated. Bioengineered silver nanoparticles (AgNPs) surface functionalized with plant extract material (proteinacious) was generated in vitro in a test tube by treating ionic AgNO3 with the leaf extract of Azadirachta indica that acted as dual reducing as well as stabilizing agent. Purity and crystallinity of the AgNPs, along with physical and surface characterizations, were evaluated by performing transmission electron microscopy, Fourier transform infrared spectroscopy, energy dispersive x-ray spectra, single-area electron diffractions, zeta potential, and dynamic light scattering measurements. Anti-corrosion studies against mild steel (MS1010) by corrosion-inducive bacterium, Bacillus thuringiensis EN2 isolated from cooling towers, were evaluated by performing electrochemical impedance spectroscopy (EIS), weight loss analysis, and surface analysis by infrared spectroscopy. Our studies revealed that AgNPs profoundly inhibited the biofilm on MS1010 surface and reduced the corrosion rates with the CR of 0.5 mm/y and an inhibition efficiency of 77% when compared to plant extract alone with a CR of 2.2 mm/y and an inhibition efficiency of 52%. Further surface analysis by infrared spectra revealed that AgNPs formed a protective layer of self-assembled film on the surface of MS1010. Additionally, EIS and surface analysis revealed that the AgNPs have inhibited the bacterial biofilm and reduced the pit on MS1010. This is the first report disclosing the application of bioengineered AgNP formulations as potent anti-corrosive inhibitor upon forming a protective layer over mild steel in cooling water towers. Graphical Abstract ᅟ.

Biosurfactant and enzyme mediated crude oil degradation by Pseudomonas stutzeri NA3 and Acinetobacter baumannii MN3.

The present study focuses on the optimization of biosurfactant (BS) production using two potential biosurfactant producer Pseudomonas stutzeri NA3 and Acinetobacter baumannii MN3 and role of enzymes in the biodegradation of crude oil. The optimal conditions for P. stutzeri NA3 and A. baumannii MN3 for biodegradation were pH of 8 and 7; temperature of 30 and 40 °C, respectively. P. stutzeri NA3 and A. baumannii MN3 produced 3.81 and 4.68 g/L of BS, respectively. Gas chromatography mass spectrometry confirmed that BS was mainly composed of fatty acids. Furthermore, the role of the degradative enzymes, alkane hydroxylase, alcohol dehydrogenase and laccase on biodegradation of crude oil are explained. Maximum biodegradation efficiency (BE) was recorded for mixed consortia (86%) followed by strain P. stutzeri NA3 (84%). Both bacterial strains were found to be vigorous biodegraders of crude oil than other biosurfactant-producing bacteria due to their enzyme production capabilities and our results suggests that the bacterial isolates can be used for effective degradation of crude oil within short time periods.

Ginger extract as green biocide to control microbial corrosion of mild steel.

In latest years, various techniques and chemicals have been used for the control of microbial influenced corrosion (MIC) of metals. The application of botanical-based biocides is one of the effective and practical techniques in the fight against MIC. In the present study, the role of aqueous extract of ginger (Zingiber officinale) (GIE) as green biocide to control MIC of mild steel 1010 (MS) in a cooling water system was investigated. Biocorrosion behavior of Bacillus thuringiensis EN2 on MS and its control by GIE was analyzed by electrochemical measurements. Polarization, electrochemical studies (ES), weight loss measurements (WL), and surface analysis (XRD, X-ray diffraction spectroscopy, and FTIR, Fourier transform infra-red spectroscopy) were performed under various incubation periods up to 4 weeks. We observed that EN2 forms a thick biofilm on the MS metal surface at the end of the incubation period and the WL significantly increased to 993 mg at fourth week when compared to the initial immersion period (194 ± 2 mg). In contrast, with addition of GIE, WL was reduced about 41 ± 2 mg over biotic system (993 ± 2 mg). GC-MS analysis confirmed the adsorption of active component of GIE (ß-turmerone) on the metal surface as a protective layer to prevent the biofilm formation and thus leads to reduction of corrosion. The optimum 20 ppm of GIE was found to be effective corrosion inhibition efficiency which was about 80%. From the results of WL, ES, XRD, FTIR, and GC-MS, GIE was identified as biocide and thus inhibits the bacterial growth on MS metal surface and it leads to control MIC. XRD showed that the GIE with EN2 resulted in less formation of corrosion products over biotic and abiotic systems. Overall, this research first shed light on the antibacterial activity of GIE inhibiting biofilm formation, thus reducing the corrosion of MS in cooling water systems.

Airborne bacteria associated with corrosion of mild steel 1010 and aluminum alloy 1100.

A novel approach to measure the contribution of airborne bacteria on corrosion effects of mild steel (MS) and aluminum alloy (AA) as a function of their exposure period, and the atmospheric chemical composition was investigated at an urban industrial coastal site, Singapore. The 16S rRNA and phylogenetic analyses showed that Firmicutes are the predominant bacteria detected in AA and MS samples. The dominant bacterial groups identified were Bacillaceae, Staphylococcaceae, and Paenibacillaceae. The growth and proliferation of these bacteria could be due to the presence of humidity and chemical pollutants in the atmosphere, leading to corrosion. Weight loss showed stronger corrosion resistance of AA (1.37 mg/cm2) than MS (26.13 mg/cm2) over the exposure period of 150 days. The higher corrosion rate could be a result of simultaneous action of pollutants and bacterial exopolysaccharides on the metal surfaces. This study demonstrates the significant involvement of airborne bacteria on atmospheric corrosion of engineering materials.

Biosurfactant and Degradative Enzymes Mediated Crude Oil Degradation by Bacterium Bacillus subtilis A1.

In this work, the biodegradation of the crude oil by the potential biosurfactant producing Bacillus subtilis A1 was investigated. The isolate had the ability to synthesize degradative enzymes such as alkane hydroxylase and alcohol dehydrogenase at the time of biodegradation of hydrocarbon. The biosurfactant producing conditions were optimized as pH 7.0, temperature 40°C, 2% sucrose and 3% of yeast extract as best carbon and nitrogen sources for maximum production of biosurfactant (4.85 g l-1). Specifically, the low molecular weight compounds, i.e., C10-C14 were completely degraded, while C15-C19 were degraded up to 97% from the total hydrocarbon pools. Overall crude oil degradation efficiency of the strain A1 was about 87% within a short period of time (7 days). The accumulated biosurfactant from the biodegradation medium was characterized to be lipopeptide in nature. The strain A1 was found to be more robust than other reported biosurfactant producing bacteria in degradation efficiency of crude oil due to their enzyme production capability and therefore can be used to remove the hydrocarbon pollutants from contaminated environment.