Bacillus Mucilaginosus Research Articles

Evaluation of thermogravimetric analysis as a rapid tool for the detection of rhizobacteria biostimulants used in precision agriculture

Six-week-old root samples were evaluated using high resolution (maximum resolution [MaxRes]) thermogravimetric analyses (TGA) of the cell wall compositions of Gram-positive (Bacillus mucilaginosus, Bacillus amyloliquefaciens, and Bacillus subtilis) and Gram-negative (Burkholderia sp., Rahnella aquatilis strain H 2.6, and R. aquatilis strain RC 2.5) root colonizing plant growth promoting rhizobacteria (PGPR) commercial inoculant strains (biostimulants) applied to pot grown wheat plants. TGA discriminated the strains within the two types of rhizobacterial cohorts and thus provided a rapid non-molecular means for the detection of PGPR inoculant biostimulants within hours of root sampling. The latter was due to the greater degree of definition of TGA fingerprints of individual thermal weight loss events occurring over a degradation range, and heightened the corresponding peak temperature divergences within strains of either type of bacteria themselves for their unequivocal identification. Confirmation of biostimulant rhizobacteria identity in concomitant root samples was achieved through either cultural methods or direct tissue PCR molecular protocols within 5 days and 2 days of sampling, respectively. The results suggested that MaxRes TGA could serve as a rapid, inexpensive stand-alone tool or as combinatorial utility alongside pyrolysis gas chromatography mass spectra, and Fourier transform infrared analytics for the early detection of PGPR biostimulants in precision farming.

Soil biochemical properties and microbial composition in aged and non‐aged apple (Malus domestica) orchards in Luochuan County, Loess Plateau, China

AbstractThe ageing of orchards is common in the apple planting process, which affects the yield and quality of apples, as well as the development of the apple industry and local economy. The investigation of soil properties is important to the improvement and soil management of aged apple orchards. In this study, we investigated the spatial distribution, correlation and clustering of soil properties (nutrients, pH, enzymes and microorganisms) in aged and non‐aged apple orchards, as well as soil microbial communities in aged orchards with and without biofertilizer (Bacillus mucilaginosus + N, P, K) in Luochuan, Loess Plateau. The results showed that most soil properties, except for pH and catalase activity, decreased gradually with depth of soil and were highest in the 0–40 cm layer in both the aged and non‐aged orchards. However, there were no obvious differences in the horizontal direction, especially in the aged orchards, indicating that the fertilizer application site was randomly located in the aged orchards. The significant correlations between soil nutrients and microbes, available phosphorus and tree age, and the clustering of orchards as aged and non‐aged groups by soil properties, indicated that soil properties should be monitored in a timely manner to assess fertilizer application and predict the ageing of orchards. Additionally, the biofertilizer might make the aged orchard soil tend to non‐ageing by increasing beneficial and reducing harmful microbes, which suggested that biofertilizers should be widely used in future fertilizer application management of aged orchards to delay ageing.

Effects of background fertilization followed by co-application of two kinds of bacteria on soil nutrient content and rice yield in Northeast China

With the improvement of living standards, people pay more and more attention to the quality and safety of rice. Microbial agents are favored by the public because they can activate the nutrient supply in the soil, and reduce the residue and application amount of chemical fertilizers and pesticides. Based on the conventional fertilization in the field, Bacillus mucilaginosus and Aspergillus niger were applied, Bacillus mucilaginosus was inoculated at four levels in the paddy soil in the cold region of Heilongjiang Province of China. The effects of different proportions of Bacillus mucilaginosus and Aspergillus niger on the number of soil microorganisms, enzyme activity, microbial biomass, soil biochemical intensity, soil nutrient content, plant nutrient content and yield were studied, and the effects on the plant nutrient content of rice and the nutrient dynamics were discussed. The results showed that a 2.62%-21.20% higher yield of rice obtained from co-application treatments compared with that of the control-blank treatment. Furthermore, the highest yield obtained (10736±65 kg/hm2) suggested that the optimized values for the two bacteria applied were 120×1011 CFU/hm2 for Bacillus mucilaginosus and 15×1011 CFU/hm2 for Aspergillus niger. Bacillus mucilaginosus can decompose minerals in soil, dissolve potassium and silicon, decompose apatite and release phosphorus into soil. Aspergillus niger can transform the phosphate which cannot be absorbed by plants into soluble phosphate which can be directly absorbed by plants by producing non-volatile acids. In particular, Bacillus mucilaginosus and Aspergillus niger have synergistic effect, and their combined application effect is greater than that of two bacteria alone. Co-application promoted the release of soil soluble silicon, and then increased the silicon content of plants. At the same time, soil microorganism, microbial biomass, enzyme activity and biochemical activity all increased significantly. This study provides an effective way to reduce the amount of chemical fertilizer applied in rice production in cold regions of China. Keywords: microbial inoculants, microbial fertilizer, rice, co-application, Bacillus mucilaginosus, Aspergillus niger, nutrients DOI: 10.25165/j.ijabe.20201302.4863 Citation: Sun T, Liu Y Y N, Wu S, Zhang J Z, Qu B, Xu J G. Effects of background fertilization followed by co-application of two kinds of bacteria on soil nutrient content and rice yield in Northeast China. Int J Agric & Biol Eng, 2020; 13(2): 154–162.

Biomineralization Performance of Bacillus sphaericus under the Action of Bacillus mucilaginosus

Microbial Induced Calcite Precipitation (MICP) is a biochemical process widely found in nature, also known as microbial mineralization. This paper investigates whether this process can help promote the intelligent reinforcement and repair of underground projects such as mines and tunnels. We selected Bacillus sphaericus and Bacillus mucilaginosus as the research objects. The former has an outstanding urease production ability, and the latter can secrete carbonic anhydrase in vitro. Bacillus mucilaginosus was introduced into the culture solution of Bacillus sphaericus in the most suitable culture environment, and the changes of mineralization rate and mineralization yield of Bacillus sphaericus were observed and analyzed. The results revealed that, to maintain the highest growth rate of Bacillus sphaericus, the optimal pH value was between 7 and 8, the optimal urea concentration was 0.5 mol/L, the optimal Ca2+ concentration was 0.6 mol/L, and the optimal Luria‐Bertani (LB) culture concentration was 20 g/L. The amount of biomineralized calcium carbonate precipitated in the double bacteria solution can reach 1.89 times the amount of the precipitation in the Bacillus sphaericus solution under the same conditions. It concludes that the introduction of Bacillus mucilaginosus can effectively increase the mineralization yield of Bacillus sphaericus without affecting the mineralized products.

Soil fertility management in apple orchard with microbial biofertilizers

Intensification of horticulture in Russia involves planting of new high-density orchards with drip irrigation and fertigation as well as intensification of the exploitation of traditional orchards. This approach involves an increase in mineral fertilizer application imposing the risk of soil fertility loss. For several reasons, the use of traditional organic fertilizers like manure in orchards is currently marginal. Although bacteria-based biofertilizers cannot substitute mineral fertilizers completely, they can significantly reduce the need for mineral fertilizer application. The effect of microbial biofertilizers of the brands “Azotovit” (Azotobacter chroococcum), “Phosphatovit” (Bacillus mucilaginosus), as well as a mixture of bacteria and the fungus, “Organic” (Azotobacter chroococcum, Bacillus subtilis, Bacillus megaterium, Trichoderma harzian) was studied in two field experiments. In the experiment #1, the preparations “Azotovit” and “Phosphatovit” were delivered through a drip irrigation system in various combinations with mineral fertilizers. In experiment #2, the preparation “Organic” was also applied to the soil with irrigation water, also in combination with the mineral fertilizer. When solely applied, none of the studied preparations changed significantly the soil nutrient content and yield as compared with the variant fertilized by the mineral fertilizer at the maximum studied application rate. The combination of the microbial biofertilizer and mineral fertilizers applied at a low rate ensured the yield commensurate to that obtained under high-rate application of the mineral fertilizer.

Rhizobacteria Effect on Arsenic Migration and Translocation of Biogenic Elements in Plants

The study was aimed at the transformation of arsenic compounds in the rhizosphere, its accumulation in plants, P and Si translocation to plants under the influence of Bacillus megaterium var. phosphaticum, and Bacillus mucilaginosus with various forms of As compounds in the soil. The authors describe the maximum effect of Bacillus megaterium var. Phosphaticum strain on As migration, its mobilization and immobilization in the rhizosphere due to arsenic leaching from mineral and difficult-todestroy compounds and its accumulation in plants. The forms of arsenic compounds were isolated from the rhizosphere based on sequential extraction procedures. The features of the inter-element As-P interaction in plants were established. With the intense accumulation of As in the rhizosphere inoculated with rhizobacteria, the intake of phosphorus into plants was not observed, as contrary to Si. The study of As and biogenic elements behavior under the influence of rhizobacteria is of great importance in the development of ecobiotechnologies related to soil remediation and crop production.

Solid-state Co-cultivation of Bacillus subtilis, Bacillus mucilaginosus, and Paecilomyces lilacinus Using Tobacco Waste Residue.

Agro-industrial wastes are excellent sources for solid-state culture to produce spores of microorganisms, whereas microbial co-cultivation is not fully exploited in solid-state culture. In this work, the co-cultivation of different strains of Bacillus subtilis, and three microbes of B. subtilis, Bacillus mucilaginosus, and Paecilomyces lilacinus was studied using a solid medium only composed of water and tobacco waste residue after extraction of nicotine and solanesol. The influences of matrix thickness, moister, temperature, and ratio of three microbes in seed on the cell growth and spore formation were studied. The maximum viable cells and spores of each microbe reached 1013 cfu/g when cultured alone at 30 °C in a medium containing 58.3% moisture. Co-cultivation of microbes stimulated cell growth and maximum viable cells of each microbe reached 1014 cfu/g, while spore production was inhibited and decreased to 1011 cfu/g. With decreasing amount of P. lilacinus in seed, total amount of spores was increased. When the seed with a ratio of 6:3:1 for B. mucilaginosus, B. subtilis, and P. lilacinus was inoculated, the total amount of spores reached 4.14 × 1012 cfu/g and the ratio was 1.7:0.7:1. These results indicate the potential of solid-state cultivation in the high production of spores from tobacco waste residue at low cost.

Blank roasting and bioleaching of stone coal for vanadium recycling

Abstract The recovery and utilization of vanadium from stone coal is of great significance because it is an important metal. In the present study, a cleaner metal extraction method to recover vanadium from low-grade stone coal using a novel strain of Bacillus mucilaginosus (BM) and an enhanced bioleaching procedure was explored. The efficient leaching bacteria of BM-5070 were obtained by the atmospheric and room temperature plasma (ARTP)-hydroxylamine hydrochloride compound mutagenesis technique. The results show that BM-5070 had the highest V leaching rate. After a 20-day bioleaching period, the V leaching rate of BM-5070 was 5.5 percentage points higher than that of the original bacteria. Moreover, the pretreatment of stone coal by blank roasting without any additives can further promote the vanadium dissolution, and the V leaching rate reached 85% after bioleaching for 20 days, which was 57 percentage points higher than that of without roasting. This is due to the decrease in the content of V (III) and the increase in the V (V) and V (IV) content after blank roasting, which is beneficial to the leaching of vanadium. The bioleaching behavior was elucidated through the bacteria-mineral contact leaching and non-contact leaching test. The results show that the V leaching rate (81.0%) in the contact mode was much higher than that in the non-contact leaching mode (64.4%), indicating that the bioleaching of vanadium occurs with direct action and indirect action. The results of our study could significantly contribute to a green method for recovering vanadium from stone coal.

Bioleaching behaviors of silicon and metals in electrolytic manganese residue using silicate bacteria

Electrolytic manganese residue (EMR) is a kind of solid waste generated along with the production of electrolytic manganese, with a high content of quartz and metals. The former can be used as silicon fertilizer for crops after activation, while the latter may cause harm to surrounding environment due to their toxicity. The aim of this study was to investigate the effects of bacterial on the activation of silicon and fixation of heavy metals in EMR by using microbe-mineral direct contact and indirect contact methods. Mixed cultures of Bacillus mucilaginosus (BM) and Bacillus circulans (BC) were used to achieve activation of silicon in EMR, and the leaching concentration of Mn, Co and Ni and their chemical forms in the leaching residue were also investigated. The results show that the microbe-mineral direct contact method has better performance for the activation of silicon in EMR with the available silicon in the leaching solution up to 163.27 mg L−1 than the indirect contact method. The metals (Mn, Co and Ni) remaining in the residue mainly exist as the stable state after bioleaching, with lower leaching concentration after bioleaching of 20 d. And the effect of silicon bacteria on activation of silicon and the fixation mechanism of metals including the effect of silicate and silicate bacteria is confirmed, and it is verified by the identification of metabolites and the analysis of chemical reaction process. This study investigates the treatment of EMR by using mixed silicate bacteria, and reveals the mechanisms of bacteria on the activation of silicon and fixaion of heavy metals in EMR. The bioleaching method not only provides silicon source for soil but also can realize the reclamation of EMR.

Improving vanadium extraction from stone coal via combination of blank roasting and bioleaching by ARTP-mutated Bacillus mucilaginosus

Abstract In order to improve leaching efficiency of vanadium from stone coal, the combination of blank roasting and bioleaching using Bacillus mucilaginosus (B. mucilaginosus) mutants was evaluated. The atmospheric and room temperature plasma (ARTP) technique was used to generate B. mucilaginosus mutants. The results showed that a mutant B. mucilaginosus BM-50, after ARTP irradiation for 50 s, had the highest acid production. The total content of the organic acid produced by B. mucilaginosus BM-50 was nearly doubled compared with the wild strain after 2 days. After 20 days, vanadium leaching rate with B. mucilaginosus BM-50 reached 18.2%, which was improved compared with the original bacteria (15.3%). A pretreatment via blank roasting for stone coal further improved the vanadium dissolution by bioleaching, namely, 68.3% vanadium was extracted, which was much higher than that without blank roasting. It is shown that bioleaching by bacterial mutants by ARTP irradiation combined with blank roasting has great potential for improving vanadium recovery from low-grade vanadium bearing stone coal.

EVALUATION USE OF BACILLUS MUCILAGINOSUS AS BIOFERTILIZER INTERFERE WITH GLOMUS MOSSEAE ON GROWTH AND YIELD OF CORN

The current study was aimed to isolate and diagnosis Bacillus mucilaginosus and evaluate its use as biofertilizer alone or interfere with Glomus mosseae on growth and yield of corn (Zea mays L.) Randomized completely block design was used at the Field of College of Agriculture- University of Baghdad Al-Jadriya in silt clay loam soil. The biofertilizer was used alone or as combination that added with or without 50 Kg P ha­¹ of rock phosphate and with or without 120 Kg K ha­¹ while, 250 Kg N ha­¹ was added as urea (46%N) to all treatments. Results showed the superiority of bacterial or fungal biofertilizer on enhancing all growth traits of corn and the addition of B . mucilaginosus together with Glomus mosseae resulted superiority for plant height , dry weight of vegetative part , grain yield and biological yield with the values 229.4 cm , 180.7 gm .plant­¹,12.17 tons ha­¹ and 19.59 tons ha­¹ respectively. As well as treatment gave significant increase equal the increase in the yield due to addition of completely recommended mineral fertilizer which was 12.14 tons ha-1 .

Активность силикатных бактерий и Bacillus mucilaginosus в дерново-подзолистой почве в отношении деградации диатомита Инзенского месторождения

Активность силикатных бактерий и Bacillus mucilaginosus в дерново-подзолистой почве в отношении деградации диатомита Инзенского месторождения

CHARACTERIZATION OF BIOFLOCCULANT-PRODUCING BACTERIA ISOLATED IN VIETNAM FOR HARVESTING OF INDIGENOUS MICROALGAE

Bioflocculation producing bacteria isolated from soil and sediment in Da Lat City, Lam Dong Province, Vietnam, were characterized. Four bacterial strains that had high flocculation activity were Flavobacterium granuli CL, Hydrogenophaga pseudoflava DA, Alcaligenes cupidus PT2, Bacillus mucilaginosus PT3, with the flocculation efficiency of 91.37%, 83.12%, 76.92%, and 75.81%, respectively at 22°C, pH 10, the initial algae concentration (optical density at 690nm) of 1.25, and grown in media containing glucose as the carbon source. The bioflocculation efficiency was depended on the pH, carbon sources, the initial algae concentrations, bacteria strains, and the inital bioflocculant-producing bacteria concentrations. The temperature lightly affected the bioflocculation, no noticeable behavioral or activity changes were observed in mice that were orally administrated with four bacterial strains, and no treatment-related illness or death occurred after 72 hours. In general, the bioflocculation process is easy to operate, cost-effective, environment-friendly and therefore, it can be applied for industrial processing of microalgae.

Influence of Biofertilizers and Liming on Vetch–Oat Mixture Productivity and Change in Sod-Podzolic Soil Microbocenosis

The influence of modification of granules of organomineral fertilizer (OMF) by BisolbiFit (Bacillus subtilis Ch-13), Phosphatovite (Bacillus mucilaginosus), and PhosphoActiv (Bacillus subtilis Ch-13 + Bacillus mucilaginosus) biopreparations on the productivity of vetch–oat mixture, as well as on changes in the structure of soil microbocenosis, has been studied under the conditions of a two-factor field experiment. The research was carried out on a soddy medium podzolic light loamy soil. The experiment included six versions: control (without fertilizers); N30P30K30; organomineral fertilizer (OMF); OMF + BisolbiFit modifier; OMF + Phosphatovite; OMF + PhosphoActive with liming (pHKCl 5.9) and without liming (pHKCl 5.1). The fertilizers were introduced with equal portions of nitrogen (30 kg a.i./ha). On average, the highest yield increase for the period from 2015 to 2017 (11.7% with application of N30P30K30 and 10.6% with application of unmodified OMF) was obtained in the OMF + BisolbiFit combined with liming version. The use of other modifiers increased the yield of the herbage of vetch–oat mixture only at the trend level. The use of all types of fertilizers increased the content of crude protein by 1.24–3.34% compared to the control. A trend towards an increase in the concentration of phosphorus and potassium in the herbage has been recorded. The highest yield increase per 1 kg of active ingredient (a.i.) of fertilizers was obtained with application of BisolbiFit: 18.0 kg of feed units without liming and 20.7 kg of feed units with liming. The modern method of high throughput sequencing of the 16S ribosomal RNA gene has detected the leading phylums, the share of which in the soil was significantly higher than that of the others: Acidobacteria (7.1–11.5%) and Actinobacteria (13.6–20.4%), Bacterroidetes (7.2–19.3%), Proteobacteria (45.3–56.2%), and Verrucomicrobia (4.3–10.3%). The highest proportion of Actinobacteria (15–20%) was recorded in versions with application of N30P30K30, OMF, and OMF + BiosolbiFit and the highest proportion of the Proteobacteria phylum was recorded in the version with application of OMF + BisolbiFit, which indicates intensive nitrification processes.

Microscopic and Spectroscopic Insights into Uranium Phosphate Mineral Precipitated by Bacillus Mucilaginosus

In this paper, we used spectroscopic and microscopic techniques to investigate the interaction mechanism between uranium and Bacillus mucilaginosus. According to scanning electron microscope couple with energy dispersive X-ray detector analysis, the lamellar uranium phosphate precipitation was only observed on the living B. mucilaginosus and the resting B. mucilaginosus. The Fourier transform infrared spectroscopy spectrum also indicated the important role of phosphate groups in forming U(VI)-phosphates precipitation. The X-ray diffraction analysis identified the phase of U(VI)-phosphate precipitation as H3OUO2PO4·3H2O. Batch experiment showed that biominerilization amount could be up to 195.84 mg/g when exposing living B. mucilaginosus to U(VI) aqueous solution at pH 5.0 for 1 h. The precipitate was further evidenced by extended X-ray absorption fine structure spectra based on the presence of U–P shell, which demonstrated that hydrogen uranyl phosphate became the main products on the living B. mucilagino...

Ranking of 38 prokaryotes according to their uranium uptake capacity in aqueous solutions: an approach from order theory through the Hasse diagram technique

ABSTRACTUranium contamination is of global concern and several approaches have been applied to cleanse U-contaminated ecosystems. Among them, biotechnological techniques using microorganisms have proven to be eco-friendly. However, knowing which of them behave better for U trapping is still an open question. In this study, a general and a detailed comparison of prokaryotes, differentiated by growth and U uptake conditions, was performed. The general comparison was run over 38 species characterized by different conditions and a subset of 18 bacteria having equal conditions was compared in detail. In both cases, the species were characterized by their percentage of U removal, uptake capacity, and time required for removing U. The methodology used was the Hasse diagram technique, suitably adjusted to address the issue of ordering objects (microorganisms) whose attributes contain interval values. Microorganisms were ranked and a stable top-five of bacteria for U trapping from aqueous systems was obtained: Bacillus licheniformis ATCC 14580, Bacillus mucilaginosus ACCC 10012, Pseudomonas MGF-48, Streptomyces sp., and Pseudomonas sp. EPS-5028. The results show that neither the Gram nor the taxonomic classification determines uptake capacity for the species studied. The method here described can be further applied to other attributes of relevance for the selection of optimal microorganisms, e.g. economical ones.

Influence of biochar application on potassium-solubilizing Bacillus mucilaginosus as potential biofertilizer

ABSTRACTBiochar can enhance soil fertility to increase agricultural productivity, whereas its improvement in soil microbial activity is still unclear. In this article, the influence of biochar on the cell growth and the potassium-solubilizing activity of Bacillus mucilaginosus AS1153 was examined. The impact on cell growth is related to the biochar-derived feedstocks and the particle size of biochar. Both intrinsic features and inner component fraction can promote the cell growth of B. mucilaginosus AS1153. The potassium-solubilizing activity was increased by 80% when B. mucilaginosus was incubated in conjunction with the biochar derived from corn stover. The survival time of B. mucilaginosus also was prolonged by adsorption in biochar. The experimental results suggested that the biochar containing B. mucilaginosus could be used as a potential biofertilizer to sustain crop production.

Preparation and partial structural characterization of the exopolysaccharide from Bacillus mucilaginosus SM-01

In this study, an acidic hetero-exopolysaccharide, designated as BMPS, was isolated from the culture supernatant of bacteria Bacillus mucilaginosus SM-01 by ion-exchange and size-exclusion chromatography. The molecular weight of BMPS was measured to be 2.67×106Da. To analyze the structure of the polysaccharide, the low-molecular weight fragment (BMPS-H) of BMPS was obtained under mild hydrolysis. BMPS-H was determined to be a liner neutral fraction with an average molecular weight of 1621Da. Monosaccharide analysis indicated that BMPS-H contained glucose and mannose in a molar ratio of 1.5:1. Based on methylation analysis and NMR spectroscopy data, the following structure of BMPS-H was established: [Display omitted] In addition, MALDI-TOF-MS analysis showed the hydroxyl group on C-2 of 1,4-linked-glucosyl residues was partially acetylated.

The Cleansing of Waste Oil in an Oilfield by Pure-culture Microorganisms

Eight strains were isolated and identified as Acinetobacter johnsonii, Bacillus cereus, Staphylococcus kloosii, Burkholderia cepacia, Serratia ficaria, Bacillus mucilaginosus, Gordonia polyisoprenivorans, and Nocardia amarae. They were employed to remove inorganic admixtures by wettability reversal in oil phase and bioflocculation in water phase. The admixtures were separated and determined as SiO2, (Na,Ca)Al(Si,Al)3O8, Al2Si2O5(OH)4, KAl2(Si3Al)O10(OH)2, CaCO3, BaSO4, and FeS. They could be effectively removed with inorganic admixture, turbidity, and sulfide below 0.5%, 100 NTU, and 2 mg/L, respectively, rendering clear oil-water interface in favor of smooth reclamations of waste oil and wash-down water.

Mitigation of efflorescence of wallboard by means of bio-mineralization.

Cement-based material is one of the most versatile and largest amounts of building materials which can not only be used in load-bearing structure but also be used as decoration materials, like brick, wallboard, and tile. However, white calcium carbonate always be found on the surface of wallboard. This phenomenon is generally called efflorescence, which has no damage to wallboard, but has aesthetic impact. In this research, Bacillus mucilaginosus was pre-added to the cement matrix to reduce the efflorescence of wallboard. Image processing, thermogravimetric analysis and permeability test were used to characterize the efflorescence degree of wallboard. The results showed that the bacterium captured atmospheric CO2 by carbonic anhydrase and promoted the CO2to react with Ca(OH)2. This process not only reduced the content of Ca(OH)2 but also improved the compactness of wallboard. In addition, the maximal decrease of efflorescence area of wallboard was gotten when the content of microbial was up to 4% of the mass of cementitious material and the proportion of efflorescence area reduced from 32 ± 3 to 5 ± 1% of the whole area of surface layer. At the same time, the values of compressive and flexural strength were the highest and the surface layer of wallboard was the most compact. The observed reduction of efflorescence was indeed due to the effect of bio-mineralization. This promising method was noted to be cheap, convenient, environment friendly, and which has the potential in various practical applications.