High Urease Activity Research Articles

Anti-seepage reinforcement property and pollution control effect of bio-cemented fracture zone in electrolytic manganese residue dump

The heavy metal (HM) pollutants in electrolytic manganese residue (EMR) can easily diffuse through the seepage channel of the dump under the leaching action of rainfall. Particularly, the fracture zone, as one of the widely distributed seepage channels in the manganese residue dump (MRD), poses a greater threat to the ecological environment due to its weak mechanical properties, strong ductility, and numerous fractures. In this study, the microbially induced calcium carbonate precipitation (MICP) method was used for anti-seepage reinforcement of the fracture zone in an MRD to be constructed. The optimal conditions for the anti-seepage reinforcement of the fracture zone using the MICP method were proposed, and the control effect of the bio-cemented fracture zone on the major HM pollutant in the leachate of EMR was illustrated. Results showed that after grouting at a rate of 3 mL/min for 12 cycles with a grouting slurry of high urease activity (9 mM urea/min) and high cementation solution concentration (1.5 mol/L), the permeability coefficient of the fracture zone decreased from 10−4 to 10−5, and the unconfined compressive strength was approximately 5.58 times that of uncemented. Furthermore, the cubic calcite crystal clusters with high purity, good stability, dense arrangement, and high cementation properties were generated using the optimal conditions, and the pores were transformed from long columnar to spherical pores. Additionally, the Mn2+ concentration in the effluent from the fracture zone at the stabilization stage was only 663 mg/L, significantly lower than that of the leachate of EMR, and the risk of leaching changed from very high risk to low risk. The research outcomes can provide guidance and reference for laboratory model testing and in-situ testing of bio-cemented fracture zone, and it is expected to provide the theoretical and technological support for green and economic anti-seepage reinforcement of the fracture zone.

Bhargavaea beijingensis a promising tool for bio-cementation, soil improvement, and mercury removal

Microbially Induced Calcite Precipitation (MICP) has emerged as a promising technique for bio-cementation, soil improvement, and heavy metal remediation. This study explores the potential of Bhargavaea beijingensis, a urease-producing bacterium, for these applications. Six ureolytic bacteria were isolated from calcareous bricks mine soil and screened for urease and calcite production. B. beijingensis exhibited the highest urease activity and calcite precipitation. Urease activity, calcite precipitation, sand solidification, heavy metal removal efficiency, and compressive strength were evaluated. It showed significant heavy metal removal efficiency, particularly highest for HgCl2. Mortar blocks treated with B. beijingensis or its crude enzyme exhibited improved compressive strength, suggesting its potential for bio-cementation. Crack remediation tests demonstrated successful crack healing in mortar blocks using the bacterium or its enzyme. This study identifies B. beijingensis as a novel and promising MICP agent with potential applications in bio-cementation, soil improvement, and heavy metal remediation. Hence, B. beijingensis diversified abilities prove superior performance compared to commonly used strains like Bacillus subtilis and Shewanella putrefaciens in bio-cementation applications. Its high urease activity, calcite precipitation, and heavy metal removal abilities make it a valuable candidate for sustainable and eco-friendly solutions in various fields.

Co-composting of dewatered sludge and wheat straw with newly isolated Xenophilus azovorans: Carbon dynamics, humification, and driving pathways

Composting is a biological reaction caused by microorganisms. Composting efficiency can be adequately increased by adding biochar and/or by inoculating with exogenous microorganisms. In this study, we looked at four methods for dewatered sludge waste (DSW) and wheat straw (WS) aerobic co-composting: T1 (no additive), T2 (5% biochar), T3 (5% of a newly isolated strain, Xenophilus azovorans (XPA)), and T4 (5% of biochar-immobilized XPA (BCI-XPA)). Throughout the course of the 42-day composting period, we looked into the carbon dynamics, humification, microbial community succession, and modifications to the driving pathways. Compared to T1 and T2, the addition of XPA (T3) and BCI-XPA (T4) extended the thermophilic phase of composting without negatively affecting compost maturation. Notably, T4 exhibited a higher seed germination index (132.14%). Different from T1 and T2 treatments, T3 and T4 treatments increased CO2 and CH4 emissions in the composting process, in which the cumulative CO2 emissions increased by 18.61–47.16%, and T3 and T4 treatments also promoted the formation of humic acid. Moreover, T4 treatment with BCI-XPA addition showed relatively higher activities of urease, polyphenol oxidase, and laccase, as well as a higher diversity of microorganisms compared to other processes. The Functional Annotation of Prokaryotic Taxa (FAPROTAX) analysis showed that microorganisms involved in the carbon cycle dominated the entire composting process in all treatments, with chemoheterotrophy and aerobic chemoheterotrophy being the main pathways of organic materials degradation. Moreover, the presence of XPA accelerated the breakdown of organic materials by catabolism of aromatic compounds and intracellular parasite pathways. On the other hand, the xylanolysis pathway was aided in the conversion of organic materials to dissolved organics by the addition of BCI-XPA. These findings indicate that XPA and BCI-XPA have potential as additives to improve the efficiency of dewatered sludge and wheat straw co-composting.

Feeding strategies for Sporosarcina pasteurii cultivation unlock more efficient production of ureolytic biomass for MICP.

The bacterium Sporosarcina pasteurii is the most commonly used microorganism for Microbial Induced Calcite Precipitation (MICP) due to its high urease activity. To date, no proper fed-batch cultivation protocol for S. pasteurii has been published, even though this cultivation method has a high potential for reducing costs of producing microbial ureolytic biomass. This study focusses on fed-batch cultivation of S. pasteurii DSM33. The study distinguishes between limited fed-batch cultivation and extended batch cultivation. Simply feeding glucose to a S. pasteurii culture does not seem beneficial. However, it was exploited that S. pasteurii is auxotrophic for two vitamins and amino acids. Limited fed-batch cultivation was accomplished by feeding the necessary vitamins or amino acids to a culture lacking them. Feeding nicotinic acid to a nicotinic acid deprived culture resulted in a 24% increase of the specific urease activity compared to a fed culture without nicotinic acid limitation. Also, extended batch cultivation was explored. Feeding a mixture of glucose and yeast extract results in OD600 of ≈70 at the end of cultivation, which is the highest value published in literature so far. These results have the potential to make MICP applications economically viable.

Response of plants and soils to inundation duration and construction of the plant‒soil association mode in the hydro‒fluctuation belt of the reservoir wetland

Hydro-Fluctuation Belt (HFB), a periodically exposed bank area formed by changes in water level fluctuations, is critical for damaging the reservoir wetland landscape and ecological balance. Thus, it is important to explore the mechanism of hydrological conditions on the plant-soil system of the HFB for protection of the reservoir wetland and landscape restoration. Here, we investigated the response of plant community characteristics and soil environment of the HFB of Tonghui River National Wetland Park (China), is a typical reservoir wetland, to the duration of inundation, as well as the correlation between the distribution of dominant plants and soil pH, nutrient contents, and enzyme activity by linear regression and canonical correlation analyses. The results show that as the duration of inundation decreases, the vegetation within the HFB is successional from annual or biennial herbs to perennial herbs and shrubs, with dominant plant species prominent and uneven distribution of species. Soil nutrient contents and enzyme activities of HFB decreased with increasing inundation duration. Dominant species of HFB plant community are related to soil environment, with water content, pH, urease, and available potassium being principle soil environmental factors affecting their distribution. When HFB was inundated for 0–30 days, soil pH was strongly acidic, with available potassium content above 150 mg kg−1 and higher urease activity, distributed with Arundo donax L., Polygonum perfoliatum L., Alternanthera philoxeroides (Mart.) Griseb., and Daucus carota L. communities. When inundated for 30–80 days, soil pH was acidic, with lower available potassium content (50–150 mg kg−1) and urease activity, distributed with Beckmannia syzigachne (Steud.) Fern.+ Polygonum lapathifolium L., Polygonum lapathifolium L., Medicago lupulina L. + Dysphania ambrosioides L. and Leptochloa panicea (Retz.) Ohwi communities. Using the constructed HFB plant-soil correlation model, changes in the wetland soil environment can be quickly judged by the succession of plant dominant species, which provides a simpler method for the monitoring of the soil environment in the reservoir wetland, and is of great significance for the scientific management and reasonable protection of the reservoir-type wetland ecosystem.

Urease from Vigna umbellata seeds: isolation, partial purification, characterization and antifungal activity

Urease (EC 3.5.1.5) hydrolyzes urea to produce ammonia and carbamate. Urease can be synthesized by plants, fungi and bacteria. In agriculture, urease is needed efficiently to play a major role in the urea cycle as source of plant nitrogen in the soil. Rice beans (Vigna umbellata) are leguminous plants from the Fabaceae family that contain high urease activity. This study isolated urease from the seed coat and without the seed coat of rice beans. The urease activity in rice beans without seed coat was higher than that of the activity with seed coat. The amount of ammonia produced with the addition of 1 µL of urease enzyme of rice beans without seed coat was 13 µg which was higher than the amino produced from the seed coat sample (8.8 µg). The concentration of urease enzyme from rice beans was obtained at 7.238 mg/mL. Analyses of gel electrophoresis indicated that urease from rice beans are composed of four polypeptide chains with molecular weights of about 52, 33, 22 and 10 kDa. Then rice beans urease exhibited no antifungal activity against Aspergillus niger, Aspergillus fumigatus, Microsporum canis, Fusarium oxysporum subsp. lini, Trichophyton rubrum, Candida glabrata and Candida albicans.

Diversity and ecological function of urease-producing bacteria in the cultivation environment of Gracilariopsis lemaneiformis

Urease-producing bacteria (UPB) provide inorganic nitrogen for primary producers by hydrolyzing urea, and play an important role in marine nitrogen cycle. However, there is still an incomplete understanding of UPB and their ecological functions in the cultivation environment of the red macroalgae Gracilariopsis lemaneiformis. This study comprehensively analyzed the diversity of culturable UPB and explored their effects on urea uptake by G. lemaneiformis. A total of 34 isolates belonging to four main bacterial phyla i.e. (Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria) were identified through 16S rRNA sequencing and were screened for UPB by urea agar chromogenic medium assay and ureC gene cloning. Our data revealed that only 8 strains contained urease. All of these UPB exhibited different urease activities, which were determined by the Berthelot reaction colorimetry assay. Additionally, the UPB strain (G13) isolated from G. lemaneiformis with higher urease activity was selected for co-culture with G. lemaneiformis to explore its role in promoting or inhibiting nitrogen uptake by macroalgae. The results showed a significant increase in urea consumption in the culture medium and the total cellular nitrogen in G. lemaneiformis in the UPB-co culture group compared to the sterile group. This suggests that the selected UPB strain positively influences nitrogen uptake by G. lemaneiformis. Similarly, isotopic assays revealed that the δ15N content of G. lemaneiformis was significantly higher in the UPB-co culture than in the control group, where δ15N-urea was the only nitrogen source in the culture medium. This indicates that the UPB helped G. lemaneiformis to absorb more nitrogen from urea. Moreover, the highest content of δ15N was found in G. lemaneiformis with epiphytic bacteria compared to sterilized (i.e. control), showing that epiphytic bacteria, along with UPB, have a compound effect in helping G. lemaneiformis absorb more nitrogen from urea. Taken together, these results provide unique insight into the ecological role of UPB and suggest that urease from macroalgae environment-associated bacteria might be an important player in marine nitrogen cycling.

Bio-cementation practice on electric-arc steel slag and comparison with natural sand

In construction projects in Khuzestan province in Iran, electric-arc steel slag (EASS) materials are used as fill materials. In this research study, bio-cementation practice on EASS materials is examined and compared with that on natural sand. For this purpose, direct shear, unconfined compressive strength (UCS) and constant head permeability tests, together with scanning electron microscopy images obtained using an instrument equipped with an energy-dispersive X-ray spectroscope, were used to analyse and compare untreated and treated materials. The results showed that despite higher urease activity and higher calcite production of EASS materials compared with natural sand, the latter showed slightly higher UCS, higher cohesion intercept enhancement and higher permeability decrease after bio-cementation treatment.

Revisiting the urease production of MICP-relevant bacterium Sporosarcina pasteurii during cultivation

The ureolytic bacterium Sporosarcina pasteurii is commonly used for Microbial Induced Calcium Carbonate Precipitation (MICP). This process has a variety of applications in the fields of construction, geotechnical engineering, and environmental remediation. However, the factors influencing urease activity of S. pasteurii during cultivation are not yet fully understood. Even though there is debate over whether higher urease activity is in fact beneficial for MICP applications, knowledge about urease production can help provide ureolytic biomass more cost-effectively, reducing overall production costs. This study revisits the effect of various cultivation conditions on growth and urease activity by using an automated high-throughput microbioreactor system combined with a novel system for high-throughput urease activity quantification. This experimental set-up allows for unprecedented data density and enables new insights into urease production of S. pasteurii.Several factors were tested, including oxygen limitation, the feeding of urea, choice of ammonium salt, the proportion of ammonium salt in the medium, and lowering the pH during cultivation; none of these had a notable impact on urease production. Only the addition of a broad spectrum of nutrients through regular feeding with highly concentrated yeast extract and glucose led to a 70 % increased specific urease activity compared to a batch culture.These results can improve the understanding of the regulatory mechanisms governing urease expression in S. pasteurii during cultivation, and allow to adapt the cultivation process accordingly. Additionally, the new system for automated high-throughput enzyme activity determination presented in this study could be applied to optimize bioprocesses involving other ion producing enzymes.

Exploring a high-urease activity Bacillus cereus for self-healing concrete via induced CaCO3 precipitation.

The structural integrity and esthetic appeal of concrete can be compromised by concrete cracks. Promise has been shown by microbe-induced calcium carbonate precipitation (MICP) as a solution for concrete cracking, with a focus on urease-producing microorganisms in research. Bacillus cereus was isolated from soil and employed for this purpose in this study due to its high urease activity. The strain exhibited strong tolerance for alkaline media and high salt levels, which grew at a pH of 13 and 4% salt concentration. The repair of concrete cracks with this strain was evaluated by assessing the effects of four different thickeners at varying concentrations. The most effective results were achieved with 10g/L of sodium carboxymethyl cellulose (CMC-Na). The data showed that over 90% repair of cracks was achieved by this system with an initial water penetration time of 30s. The study also assessed the quantity and sizes of crystals generated during the bacterial mineralization process over time to improve our understanding of the process. KEY POINTS: • MICP using Bacillus cereus shows potential for repairing concrete cracks. • Strain tolerates alkaline media and high salt levels, growing at pH 13 and 4% salt concentration. • Sodium carboxymethyl cellulose (CMC-Na) at 10g/L achieved over 90% repair of cracks.

Garlic extract addition for soil improvement at various temperatures using enzyme-induced carbonate precipitation (EICP) method

Enzyme-induced carbonate precipitation (EICP) is an emerging technique to improve the soil and most studies are carried out at room temperature. However, considering some foundations are in high-temperature environments (>40 °C), the higher urease activity at high temperature results in the solidification inhomogeneity, limiting the application of EICP. The higher urease activity at high temperature hampers the application of EICP because of solidification inhomogeneity. The garlic extract has been used as a type of urease inhibitor in medical science and food engineering. Here, we propose to use it to control urease activity for sand solidification at high temperature. The effects of garlic extract addition on urease activity and precipitation rates for calcium carbonate (CaCO3) were studied. Extra tests were conducted to study the effect of garlic extract addition on the solidification homogeneity. The results showed that the garlic extract addition significantly decreased urease activity. To reduce the rate of CaCO3 precipitation at different temperatures, a suitable concentration of garlic extract was necessary to obtain a suitable urease activity. In the sand solidification test, garlic extract addition resulted in a smaller difference in sonic time values or CaCO3 contents at different parts of samples. The improved solidification homogeneity can achieve higher strength. The correlation between sonic time values and CaCO3 content was higher than that between CaCO3 content and strength. Appropriate concentrations of garlic extract were obtained at 35 °C, 40 °C, 45 °C, 50 °C, and 55 °C. The proposed garlic extract addition method was significant to improve the homogeneity of solidified soil in practical engineering applications.

Ecological restoration of tidal flat and its effects on soil characteristic and bacterial community

AbstractVegetation restoration is a vital ecological remediation technology for greening saline‐alkaline soils. However, little attention is paid to the seasonal changes of soil remediation under different vegetation coverage. This study investigated a recently restored coastal tidal flat located on the island of Zhoushan, by examining shifts in physicochemical properties, enzyme activities, and bacterial communities over 1 year. Soils under Suaeda australis (SUA), Phragmites australis (PHR), Tamarix chinensis (TAM), and two hybrid treatments (SUA+PHR and SUA+TAM) were collected seasonally. Results showed that the seasonal climate variations and vegetation types played important roles in soil physicochemical properties and soil enzyme activities during vegetation restoration. Vegetation could stimulate the activities of soil microbial enzymes, of which PHR‐covered soils exhibited higher catalase activities, higher alkaline phosphomonoesterase activities were observed in summer except for PHR. The combined communities had the highest urease activities in spring and summer, and the highest levels of polyphenol oxidase activity were found in summer or autumn. Furthermore, the bacterial alpha diversity revealed that SUA‐ and SUA+PHR‐covered soils were not markedly affected as the season progressed, whereas PHR and SUA+TAM were characterized by an increase–decrease trend, as well as TAM exhibited highest in autumn. Proteobacteria, Bacteroidetes, Acidobacteria, Actinobacteria, and Chloroflexi were the dominant phyla in each vegetation types. This study deepens our understanding of the effects of halophyte cultivation on coastal ecosystem phytoremediation.

Experimental study on the calcium carbonate production rates and crystal size of EICP under multi-factor coupling

The production and microstructure of calcium carbonate are the fundamental indicators to evaluate the effectiveness of Enzyme-induced Carbonate Precipitation (EICP). Soybean urease is extracted, and the urease activity is controlled by the soybean powder concentration in this study. The urease reacts with the cementation solution with different concentrations of urea and calcium ions at different temperatures to determine the calcium carbonate production rates (RCC). The crystal sizes of calcium carbonate (SCC) are investigated by scanning electron microscopy (SEM) and image processing method. The RCC increase and subsequently decrease with increasing calcium ion concentration and peak at 0.75 M. A lower urea concentration is more favourable to the formation of CaCO3. Increased urease activity and temperature will lead to an increased RCC and SCC. Some crystals appear as capsules when the temperature goes up. The SCC decreases slightly with increasing calcium ion concentration under low urease activity and increases under high urease activity. The SCC increases first and subsequently decreases with increasing urea concentration under high urease activity, while the effect of urea concentration under low urease activity is not significant. XRD test results show that the calcium carbonate produced in this study is dominated by calcite crystals. The study provides additional data, a useful base to move forward toward a better understanding of the method of stabilization and its applications, for example, the treatment of different soils.

Physical, chemical and microbiological soil attributes influence soil greenhouse gases fluxes in Atlantic Forest and pine (Pinus taeda) plantations in Brazil

AbstractForest soils can be sources or sinks of greenhouse gases (GHGs) depending on soil attributes that affect biomass and activity of soil micro‐organisms involved in GHGs fluxes. In this work, we tested the hypothesis that soil physical, chemical and microbiological attributes, under different forests ecosystems, affect the soil GHGs [nitrous oxide (N2O), carbon dioxide (CO2) and methane (CH4)] fluxes. The study was carried out in two locations in southern Brazil in 2019, with three experimental plots of 900 m2 in native forests of the Atlantic Forest biome and in loblolly pine (Pinus taeda) plantations. Air samples released from the soil surface were analysed for concentration and flux of CO2, N2O and CH4. Soil samples were analysed for chemical attributes, density (Ds), soil microporosity (MiPs), soil macroporosity (MaPs), total porosity (TP), water‐filled pore space (WFPS), microbial biomass carbon (MB‐C), basal respiration (BR), microbial (qMic) and metabolic (qCO2) quotient and activities of soil urease and β‐glucosidase enzymes. The seasons influenced the CO2 and N2O emissions, probably because of the changes in seasonal conditions. However, native forests consumed more CH4 than pine plantations. Meanwhile, the native forests presented soils with lower Ds (average 21.5% lower), more TP (average 12.5% higher) and more moisture (average 33% higher), which improved the microbiological attributes of the soil (20% to 60% more MB‐C, 67% higher urease activity and 30% higher β‐glucosidase activity) compared with pine plantations. Native forests contributed more intensely to CH4 consumption than pine plantations because they present better physical, chemical and microbiological soil conditions. Therefore, it is possible that forestry practices that improve soil physical attributes are likely to contribute to increase CH4 consumption, and to reduce GHGs emissions in forest ecosystems.

Long-Term Organic Manure Application Alters Urease Activity and Ureolytic Microflora Structure in Agricultural Soils

Ureolytic microbes in soil produce urease to catalyze the hydrolysis of urea to NH3/NH4+. Manure is widely applied in agriculture and has the potential to influence soil urease activity. In this study, we examined the responses of the ureolytic microbial community to manure application in two agricultural soils from north (N) and south (S) China using high-throughput sequencing of the ureC genes. We found that N soil and S soil harbored significantly distinct ureolytic communities, as no OTU was shared between two locations. The slight variation of the ureolytic community (32.2%, Adonis) was observed in N soil where low rates of manure were applied. However, dramatic alteration of the structure of ureolytic community (83.4%, Adonis) was found, possibly by promoting the growth of Betaproteobacteria and Deltaproteobacteria and the suppression of the growth of Actinobacteria in S soil where high rates of manure were inputted. The total C and C/N ratio were the main environmental factors driving the microbial communities. The relative ratios of ureC to 16S rRNA genes ranged from 1.5 to 3.5% among the two soils. The abundance of ureC genes was significantly and positively correlated with total phosphorus (TP, r = 0.87, p < 0.001). Positive correlations between the urease activity and soil available NH4+ (r = 0.81, p = 0.001), TP (r = 0.84, p = 0.001), and the abundance of ureC (r = 0.87, p < 0.001) were observed in our study. We speculate that sufficient soil phosphorus promotes the growth of ureolytic microbes, which results in higher urease activity and the greater release of available NH4+.

1H NMR metabolic profiling of Staphylococcus pseudintermedius isolated from canine uroliths.

Staphylococcus pseudintermedius is a urease-producing bacteria which is a major cause of magnesium ammonium phosphate (MAP) urolithiasis in canine. A positive urolith culture is an important risk factor for MAP urolithiasis in canine. The mechanism underlying the metabolic changes of S. pseudintermedius after crystallization in artificial urine (AU) needs more defined baseline metabolic information. Therefore, we extensively investigated the metabolic changes of S. pseudintermedius extensively after crystallization in AU. A high urease activity and positive biofilm formation strain, entitled the S. pseudintermedius (SPMAP09) strain, was isolated from canine MAP uroliths, and analyzed using nuclear magnetic resonance (NMR) spectroscopy-based metabolomics. The molecular mechanism-specific metabolic phenotypes were clearly observed after crystallization in AU at day 3. The crystals induced by SPMAP09 were also confirmed and the major chemical composition identified as struvite. Interestingly, our findings demonstrated that a total of 11 identified metabolites were significantly changed. The levels of formate, homocarnosine, tyrosine, cis-aconitate, glycolate, ethyl malonate, valine and acetate level were significantly higher, accompanied with decreased levels of inosine, glucose, and threonine at day 3 compared with the initial time-point (day 0). In addition, our results exhibited that the glyoxylate and dicarboxylate metabolism was significantly related to the SPMAP09 strain at day 3 in AU. Thus, metabolic changes of the SPMAP09 strain after crystallization in AU potentially helps to explain the preliminary molecular mechanism for the crystals induced by S. pseudintermedius.

The host niches of soybean rather than genetic modification or glyphosate application drive the assembly of root-associated microbial communities.

Plant roots significantly influence soil microbial diversity, and soil microorganisms play significant roles in both natural and agricultural ecosystems. Although the genetically modified (GM) crops with enhanced insect and herbicide resistance are thought to have unmatched yield and stress resistance advantages, thorough and in-depth case studies still need to be carried out in a real-world setting due to the potential effects of GM plants on soil microbial communities. In this study, three treatments were used: a recipient soybean variety Jack, a triple transgenic soybean line JD321, and the glyphosate-treated JD321 (JD321G). Three sampling stages (flowering, seed filling and maturing), as well as three host niches of soybean rhizosphere [intact roots (RT), rhizospheric soil (RS) and surrounding soil (SS)] were established. In comparison to Jack, the rhizospheric soil of JD321G had higher urease activity and lower nitrite reductase at the flowering stage. Different treatments and different sampling stages existed no significant effects on the compositions of microbial communities at different taxonomic levels. However, at the genus level, the relative abundance of three plant growth-promoting fungal genera (i.e. Mortierella, Chaetomium and Pseudombrophila) increased while endophytic bacteria Chryseobacterium and pathogenic bacteria Streptomyces decreased from the inside to the outside of the roots (i.e. RT → RS → SS). Moreover, two bacterial genera, Bradyrhizobium and Ensifer were more abundant in RT than in RS and SS, as well as three species, Agrobacterium radiobacter, Ensifer fredii and Ensifer meliloti, which are closely related to nitrogen-fixation. Furthermore, five clusters of orthologous groups (COGs) associated to nitrogen-fixation genes were higher in RT than in RS, whereas only one COG annotated as dinitrogenase iron-molybdenum cofactor biosynthesis protein was lower. Overall, the results imply that the rhizosphere host niches throughout the soil-plant continuum largely control the composition and function of the root-associated microbiome of triple transgenic soybean.

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.

Strain Screening and Particle Formation: a Lysinibacillus boronitolerans for Self-Healing Concrete.

Microbial-induced calcite precipitation is a promising technology to solve the problem of cracks in soil concrete. The most intensively investigated microorganisms are urease-producing bacteria. Lysinibacillus that is used as urease-producing bacteria in concrete repair has rarely been reported. In this study, Lysinibacillus boronitolerans with a high urease activity was isolated from soil samples. This strain is salt- and alkali-tolerance, and at pH 13, can grow to ~OD600 2.0 after 24 h. At a salt concentration of 6%, the strain can still grow to ~OD600 1.0 after 24 h. The feasibility of using this strain in self-healing concrete was explored. The data showed that cracks within ~0.6 mm could be repaired naturally with hydration when spores and substrates were added to the concrete in an appropriate proportion. Moreover, the number and morphology of CaCO3 crystals that were produced by bacteria can be influenced by the concrete environment. An efficiency method to elucidate the process of microbial-induced calcium carbonate crystal formation was established with Particle Track G400. This study provides a template for future studies on the theory of mineralization based on microorganisms. IMPORTANCE The formation of calcium carbonate crystals in concrete by urease-producing bacteria is not understood fully. In this study, a Lysinibacillus boronitolerans strain with a high urease activity was isolated and used to analyze the counts and sizes of the crystals and the relationship with time. The data showed that the number of crystal particles increases exponentially in a short period with sufficient substrate, after which the crystals grow, precipitate or break. In concrete, the rate-limiting steps of calcium carbonate crystal accumulation are spore germination and urease production. These results provided data support for the rational design of urease-producing bacteria in concrete repair.

Biocalcifying Potential of Ureolytic Bacteria Isolated from Soil for Biocementation and Material Crack Repair.

Microbially induced calcium carbonate precipitation (MICP) has been highlighted for its application in civil engineering, and in the environmental and geotechnical fields. Ureolytic activity is one of the most promising bacterial mechanisms in terms of inducing calcium carbonate formation. In this study, four bacterial isolates with high-yield urease production capabilities were obtained from two-step screening using a high-buffered urea medium. The highest urease activity and calcium carbonate formation was observed in Lysinibacillus fusiformis 5.1 with 4.40 × 103 unit/L of urease and 24.15 mg/mL of calcium carbonate, followed by Lysinibacillus xylanilyticus 4.3 with 3.93 × 103 unit/L of urease and 22.85 mg/mL of calcium carbonate. The microstructure of the precipitated crystalline calcium carbonate was observed using scanning electron microscopy. X-ray diffraction analysis confirmed that the main polymorph of the calcium carbonate particle obtained from both isolates was calcite. Examination of the material-crack filling in mortar specimens showed that calcite layers had formed along the crack edges and inside after 10 days, and gradually filled the cracks up to the upper surface. These results showed that these two isolates presented robust characteristics of potential MICP-inducing bacteria for civil engineering and material engineering applications.