Isolation and Screening of Hydrocarbon Degrading Bacterial Strains for Bioremediation of Petroleum Pollution in Qatar

Pollution, due to activities related to the oil industry, represents a serious threat to the natural environment. The application of biotechnological methods provides much safer and sustainable alternatives for bioremediation of polluted areas, using microorganisms. Several techniques for the isolation of hydrocarbon degrading bacteria have been investigated and published worldwide. A wide range of bilogical activities was shown. However, local hydrocarbon degrading strains and the factors affecting bacterial and strains variability were not studied deeply. In this study, we showed that the isolation and screening strategy affected a lot, the selection of the strains. We combined the bacterial tolerance to hydrocarbon toxicity, assessed by the growth parameters, and the bacterial degradative activities, assessed by the degradation of a wide range of petroleum hydrocarbons via Gas Chromatography analysis. The main investigations and findings of the present work are: -A collection of 39 bacterial isolates from the Qatari environment was set up and a new isolation and screening program was proposed. -The Growth conditions and the activity of pre-selected strains -Shift of the activity of the selected strains from a range of hydrocarbons to another by the effect of the nitrogen source, C/N ratio and organic nitrogen source -70% hydrocarbon removal, achieved with several strains in 2 weeks. -Amongst 12 identified isolates and by molecular ribotyping-DNA sequencing, 3 Pseudomonas strains isolated from a polluted area in Qatar, are available for bioremediation of highly polluted soils, tolerating high toxicity and may be adapted to a variety of low or high molecular weight hydrocarbons

Influence of temperature, salinity and Mg:Ca ratio on microbially-mediated formation of Mg-rich carbonates by Virgibacillus strains isolated from a sabkha environment.

Studies have demonstrated that microbes facilitate the incorporation of Mg into carbonate minerals, leading to the formation of potential dolomite precursors. Most microbes that are capable of mediating Mg-rich carbonates have been isolated from evaporitic environments in which temperature and salinity are higher than those of average marine environments. However, how such physicochemical factors affect and concur with microbial activity influencing mineral precipitation remains poorly constrained. Here, we report the results of laboratory precipitation experiments using two mineral-forming Virgibacillus strains and one non-mineral-forming strain of Bacillus licheniformis, all isolated from the Dohat Faishakh sabkha in Qatar. They were grown under different combinations of temperature (20°, 30°, 40 °C), salinity (3.5, 7.5, 10 NaCl %w/v), and Mg:Ca ratios (1:1, 6:1 and 12:1). Our results show that the incorporation of Mg into the carbonate minerals is significantly affected by all of the three tested factors. With a Mg:Ca ratio of 1, no Mg-rich carbonates formed during the experiments. With a Mg:Ca ratios of 6 and 12, multivariate analysis indicates that temperature has the highest impact followed by salinity and Mg:Ca ratio. The outcome of this study suggests that warm and saline environments are particularly favourable for microbially mediated formation of Mg-rich carbonates and provides new insight for interpreting ancient dolomite formations

Mapping of aeolian deposits of an industrial site in the arid region using the TIR bands of ASTER and study of physicochemical characters and stabilization of sand erosion

This study describes the spectral emissive character of silicate and carbonate minerals of aeolian deposits and maps the deposits and sand encroachments that occurred in and around the site 5/6 of Qatar Fertilizer Company (QAFCO), Qatar using thermal infrared (TIR) bands of ASTER. The results of studies show that the quartz and unaltered silicates have spectral features between 8.12 and 9.27 µm, and the calcite and dolomite have emissivity minima near 11.4 and 11.2 µm, respectively. The mapping of deposits, dunes, and carbonate formations using the bands, and quartz index (QI), and carbonate index (CI) displayed their occurrence, distribution, and direction of sand movement from NW to SE. The sand encroachment was mapped using high spatial resolution satellite data of WorldView-2. The study of physicochemical characteristics of field samples showed the occurrence of sand grains up to 99.81% and the XRD and geochemical analyses represented the presence of quartz, calcite, dolomite, albite, and halite minerals in the deposits. In addition, the bacterial strains isolated from the samples indicated high urease activity leading to precipitation of carbonate minerals via microbially induced calcite precipitation (MICP) processes, and demonstrated high potential for utilization for sand stabilization of the QAFCO site

Microbial Mats from the Khor Al-Adaid Sabkha, Qatar: Morphotypes and Association with Authigenic Minerals

The sabkhas (i.e., salt flats) of Qatar are among the rare places on Earth where carbonate and sulfate minerals similar to those constituting economically important hydrocarbon reservoirs are still forming today, under the arid conditions that characterize the coastline of the country. Since the 1960's, the sabkhas of Qatar have been studied with great interest as a modern analogue for ancient sedimentary sequences (e.g., Wells, 1962; Illing & Taylor, 1995; Alsharhan & Kendall, 2003). The results of these studies provided important insights for formulating stratigraphic models of subsurface hydrocarbon reservoirs. Notable examples of gas and oil reservoirs that formed in arid, evaporitic environments include the Permo-Triassic Khuff (which is estimated to contain about 15-20% of the world's gas reserves and is of fundamental importance for the economy of Qatar), the Jurassic Arab formations, and the Triassic Kurra Chine, all of the Middle East, and the Permian Zechstein of Northern Europe. Although extremely valuable, most of these early studies were based on purely physical and chemical approaches, which may have not fully captured the complexity of the mineralization processes occurring in the sabkha environment. Indeed, research conducted in more recent years has shown that microorganisms play an important and, as yet, poorly understood role for the mineralization processes occurring in these evaporitic environments (Bontognali et al., 2010; Bontognali et al., 2012; Bontognali et al., 2014; Brauchli et al., 2015; Paulo & Dittrich, 2013; Strohmenger et al., 2011). Here we present the results of a field campaign conducted in the Khor Al-Adaid sabkha, which is located in the southeast of Qatar, in a large tidal embayment composed of two shallow inland lagoons. The main goal of the field campaign was to identify regions of the intertidal zone that are particularly rich in microbial mats, and that represent ideal sites at which to study microbe-mineral interactions. Three sites of interest have been defined.qscienc

Mechanistic insights into the remediation of bromide ions from desalinated water using roasted date pits

Bromide ions (Br−) are associated with toxic by-products that might be produced during the disinfection of drinking water. The removal of Br− from water using roasted date pits (RODPs) and activated carbon was investigated. The percentage removal capacity of Br− was studied under various experimental conditions including solution pH, adsorbent mass and particle size, and initial Br− concentration. Surface characterisation was investigated using scanning electron microscopy (SEM) and Fourier transform infrared (FTIR). SEMs images showed the presence of fine particles (debris) on the surface of the adsorbent solids. The raw date pits (RDPs) surface images showed a marked difference in pore sizes upon roasting, whilst FTIR spectra showed a slight shifting in –O–H, Cdouble bondO, and –C–O stretching in addition to –O–H bending for the RODPs. In addition to the Freundlich and Langmuir isotherm models, the experimental data were fitted to different adsorption kinetic models. It was found that the adsorption of Br− onto RODPs followed pseudo-second order mechanism. Removal rates of around 54% were obtained at lower pH.This publication was made possible by UREP grant # (UREP 14-062-1-008 ) from the Qatar National Research Fund (a member of Qatar foundation). The statements made herein are solely the responsibility of the authors. We would like to offer our thanks to Sana Khan, Maymoona Ayesh, Fatima Ferhai for their contribution to some of the experimental work.Scopu

Evaluating the role of dissolved silica for dolomite formation in evaporitic environments: Insights from prolonged laboratory experiments

The mineral Dolomite CaMg(CO3)2 is a common constituent of sedimentary rocks. Despite centuries of research, the mechanism of its formation remains elusive and debated. Recent studies have shown the presence of silica in solution promote the incorporation of Mg into the carbonate mineral, forming crystal phases that may be precursors to dolomite. The goal of this study was to evaluate with laboratory experiments whether dissolved silica may play a role for dolomite formation in sabkha (i.e., salt flats) environments. Several models for dolomite formation are based on the studies of sabkhas, which are often cited as modern analogue for ancient dolomite-rich sedimentary sequences. We performed long-incubation time (i.e., up to 600 days) laboratory precipitation experiments at 30 °C with solution mimicking the sabkha pore waters (characterized by a salinity of 23 % and Mg: Ca ratio of 15) to which we added different concentrations of Si. Our results revealed a positive correlation (p-value <0.001) between Si concentration in solution and the mol% Mg of the carbonate minerals forming in the experiment. With 2 mM of Si, the bulk precipitate was comprised of 90 % stoichiometric dolomite with possible signs or ordering. Moreover, the rhombohedral morphology of the crystals is analogue to that of natural dolomite previously described from sabkha sediments. Our results suggest that dissolved Si may play an important role for dolomite formation in evaporitic environments.Scopu

Microbially influenced formation of anhydrite at low temperature

Calcium sulfate minerals are abundant in nature – on Earth and on Mars – and important in several fields of material sciences. With respect to gypsum and bassanite, anhydrite represents the anhydrous crystalline phase in the CaSO4–H2O system. Despite years of research, the formation of anhydrite in the laboratory at low temperature remains challenging and, in the geological record, this mineral is mostly interpreted as a secondary phase that form through metamorphic dehydration of gypsum. Here, we present the results of laboratory precipitation experiments showing that anhydrite can form at 35 °C from evaporated seawater through a microbially influenced mineralization process. The experiments were conducted in the presence of extracellular polymeric substances (EPS) produced by bacterial strains isolated from a modern evaporitic environment, the Dohat Faishakh sabkha in Qatar. Without organic molecules, only gypsum formed in parallel control experiments. This finding provides a possible explanation for the origin of several natural occurrences of anhydrite that cannot be satisfactorily explained by existing models and reveals a new precipitation pathway that may have industrial applications.This publication was made possible by the grant NPRP13S-0207-200291 from the Qatar National Research Fund (a member of the Qatar Foundation). The statements made herein are solely the responsibility of the authors

Characterization of the extracellular polymeric substances (EPS) of Virgibacillus strains capable of mediating the formation of high Mg-calcite and protodolomite

The origin of dolomite -a common mineral in the geological record- is the subject of an ongoing debate. Among different hypotheses, it has been proposed that extracellular polymeric substances (EPS) excreted by microbes include organic molecules that catalyze the incorporation of Mg in the carbonate mineral. However, limited information exists on the composition of the EPS produced by Ca-Mg carbonate-forming microbes, which in turn hampers a precise understanding of their role in the mineralization mechanism. Here, we present the results of laboratory experiments in which we cultured different strains of microbes, characterized their EPS, and identified components associated with carbonate minerals with high mol% Mg. Two Virgibacillus strains known to mediate the formation of Mg-rich carbonates, as well as a strain of Bacillus licheniformis -a negative control that does not mediate mineral formation but produces EPS, were grown under different salinities and temperatures, which caused them to produce EPS with different compositions. The EPS were subsequently characterized by measuring total carbohydrate (TCHO) and total protein (TP) contents, as well as by Fouriertransform infrared spectroscopy (FTIR). At the tested conditions, we found that Mg-carbonates with a mol% Mg higher than 40% (i.e., potential dolomite precursor phases) formed exclusively in association with EPS rich in carbohydrates (TCHO > than 75% of the total mass). FTIR spectra of Ca-Mg carbonate-forming strains were distinct from those of the non-mineral-forming strain in areas associated with the protein structures responsible for the formation of hydrogels, which contribute to hydration or dehydration of ionic clusters; further differences have been observed in the regions of phosphoryl functional groups. These results provide insight on which fraction of organic molecules and specific functional groups are, among the many constituents of EPS, important for mineral nucleation and incorporation of Mg into carbonate minerals, a crucial step for the formation of dolomite in natural environments

Systematic laboratory approach to produce Mg-rich carbonates at low temperature

Dolomite is a common Mg-rich carbonate in the geological record, but the mechanism of its formation remains unclear. At low temperature, the incorporation of magnesium ions into the carbonate minerals necessary to form dolomite is kinetically inhibited. Over the decades, several factors that possibly allow for overcoming this kinetic barrier have been proposed, and their effectiveness debated. Here, we present the results of a large number of laboratory precipitation experiments that have been designed to identify and compare the factors that promote the formation of Mg-rich carbonates. Under the tested conditions, the most interesting observations include: (1) from solutions that mimic evaporitic seawater, the maximum mol% of Mg incorporated in high Mg calcite is 35, (2) carbonates with a mol% of Mg above 40 were obtained exclusively in the presence of organic molecules, (3) no correlation was observed between the charge of the organic molecules and the incorporation of Mg, (4) the mode (i.e., slow vs. fast mixing) used to add carbonate to the solution obtaining supersaturation has a significant impact on the forming mineral phase (aragonite vs. nesquehonite vs. high Mg calcite) and its Mg content. These findings allow for a more informed evaluation of the existing models for dolomite formation, which are based on the study of natural environments and ancient sedimentary sequences.This work was made possible by the grants NPRP10-0214-170462 and NPRP13S-0207-200291 from the Qatar National Research Fund (a member of the Qatar Foundation)

Variability of blue carbon storage in arid evaporitic environment of two coastal Sabkhas or mudflats

Abstract Coastal Sabkhas are mudflats found in arid coastal regions that are located within the supratidal zone when high rates of evaporation lead to high salinity. While evaporitic minerals often accumulate underneath the surface, the microbial mats are present on the surface of Sabkhas. Coastal Sabkha, an under-studied ecosystem in Qatar, has the potential to store blue carbon. In the present study, we investigated the carbon storage capacity of two Sabkhas from contrasting geological backgrounds. The spatial and temporal variabilities of the carbon stocks were examined. The results showed that both studied Sabkhas exhibit a considerable potential for soil carbon storage with carbon stocks of 109.11 ± 7.07 Mg C ha−1 and 67.77 ± 18.10 Mg C ha−1 in Dohat Faishakh and Khor al Adaid Sabkha respectively. These values fall within the reported range for carbon stocks in coastal Sabkhas in the region (51–194 Mg C ha−1). Interestingly, the carbon stocks in the sediments of the Sabkhas were higher than those in the sediments of Qatari mangroves (50.17 ± 6.27 Mg C ha−1). These finding suggest that coastal Sabkhas can serve as blue carbon ecosystems in arid environments