Biological cementing mixtures for dust control and related applications

Abstract

The present invention relates mainly to the use of a mixture for reducing dust formation and / or erosion. The invention also relates to a method for reducing dust formation and / or erosion, and to a mixture suitable for this purpose.

Description

Technical Field

[0001] This invention relates primarily to the use of mixtures for reducing dust formation and / or erosion, and also to mixtures suitable for this purpose. Other aspects, and in particular other uses, will be apparent from the description below. Background Technology

[0002] Dust is virtually everywhere. To a large extent, it originates from unpaved roads (streets, railways, airports), agricultural land, mining (including open-pit mining), construction sites, industrial land, landfills, and more. Continuous dust exposure has a number of drawbacks; for example, dust is harmful to health and can potentially lead to hay fever, allergies, or pneumoconiosis. Dust harms the environment; it transports and disperses toxic substances such as chemicals, heavy metals, viruses, and microorganisms. Dust can impair traffic when it obstructs the vision of road users. Simply removing material from surfaces will create dust. Unnecessary material removal also leads to unnecessary material loss. Dust can accumulate inside and on machinery (such as vehicles and construction equipment) and can cause damage and shorten maintenance cycles, especially for moving parts.

[0003] Therefore, various dust suppression measures have been proposed in the existing technology.

[0004] These measures include covering with tarpaulins and planting vegetation. However, both methods are often costly and not always feasible. For example, planting vegetation on traffic roads such as highways is not practical.

[0005] Another suggestion is to increase the moisture content in the ground / matrix, for example, by adding water, salt water, etc. (Naeimi M, Chu J, Environmental Science and Pollution Research 24.29.2017.23341-23350; Mayer, FD et al., Earth Science Frontiers 2011: Advances in Geotechnical Engineering.2011.4002-4011; brief introduction of each literature). The disadvantage of this technique is that the effect only lasts until the moisture evaporates. Therefore, in hot and dry climates, the duration of this measure may be short. To achieve a lasting or even permanent effect, repeated application may be necessary, which is laborious and costly. Another disadvantage is that the proposed salt solution is highly corrosive to metals (and therefore vehicles and machinery) and concrete. Furthermore, since the salt solution enters the soil and groundwater, this technique is undoubtedly harmful to the environment.

[0006] An alternative method for dust control is suggested to be the use of distillation residue (WO 2009 / 151316) or coal tar pitch (EP0 305 621). However, both of these substances are toxic. Therefore, their dissemination is unacceptable for environmental protection purposes.

[0007] EP 2 838 969 recommends the use of polymer dispersions for dust suppression. However, a disadvantage of using polymer dispersions is that they are generally non-biodegradable or poorly biodegradable.

[0008] According to other methods, organic compounds such as lignin, lignin sulfonates, surfactants, polyacrylamide, starch ethers, polyacrylonitrile, polyvinyl alcohol, polyacrylamide, carboxymethyl cellulose, and polyvinyl acetate are used to bind dust (Stabnikov V et al., Water, Air, Soil Pollution. 2013. 224: 1631, Introduction). The biggest drawback stated in the prior art is the high cost, especially in the case of widespread application (Stabnikov V et al., above, Table 2).

[0009] The microbial bio-adhesive described in WO 2006 / 066326 at least partially overcomes the aforementioned drawbacks. The disclosed specification reveals a method for forming a high-strength bio-adhesive in a permeable starting material, wherein the starting material is mixed with effective amounts of (i) urease-producing microorganisms, (ii) urea, and (iii) calcium ions. Urease catalyzes the conversion of urea into carbonate, and its reaction with the provided calcium ions forms calcium carbonate, thus consolidating the starting material. The gist of the partial description is that the disclosed method is applicable to applications in mining, construction engineering, or providing specialized materials. Compared to the conventional techniques described above, bio-adhesives can partially or completely eliminate the use of non-biodegradable substances. There is no risk of damage to machinery, vehicles, etc., and the application cost is lower (Stabnikov V et al., above, Table 2).

[0010] As an aspect related to dust formation, soil erosion refers to the excessive erosion of soil by wind and water, particularly following human misuse of land, such as overgrazing, deforestation leading to the clearing of protective vegetation, and insufficient fallow periods. In this context, a specific problem is the loss of the most fertile and agriculturally significant topsoil. Persistent soil erosion initially leads to soil degradation (soil deterioration). Degradation can ultimately result in the complete loss of the soil's agricultural utility (soil destruction). Soil erosion has profound environmental, economic, and social consequences. Consequently, various soil conservation measures have been initiated worldwide, but the problem has not yet been completely eliminated. Summary of the Invention

[0011] Therefore, a primary objective of the present invention is to provide measures for reducing dust and / or erosion that at least partially overcome the aforementioned drawbacks, and whose dust and / or erosion reduction is more refined and durable compared to known biocementing. Another objective of the present invention is to provide measures for reducing dust and / or erosion that better maintain their dust and / or erosion-reducing properties after mechanical exposure, thus making surfaces treated in this manner equally usable. Further objectives of the invention (additionally or alternatively) become apparent from a study of the specification, claims, and particularly the embodiments.

[0012] According to the present invention, this or these objectives can be achieved by means of the use of mixtures for reducing dust formation and / or erosion, as described in more detail herein, and by means of mixtures suitable for this purpose.

[0013] Other aspects and preferred configurations of the invention will become apparent from the following observations, the appended embodiments, and especially from the appended claims.

[0014] According to the present invention, the mixtures described herein are suitable for bio-binding. In the context of this invention, the term "bio-binding" refers to the consolidation and / or hardening of a (permeable) matrix (as defined below in the context of the methods of the present invention). Thus, on / among these matrices, dust formation is prevented or reduced. In the context of this invention, a bio-binding agent is a product of bio-binding as defined herein.

[0015] Assume that the consolidation and / or hardening constitutes a process in which portions of the matrix to be reduced or prevented from forming dust are joined together by one or more adhesives present in and / or formed by components of the mixture used according to the invention, and in this way, the matrix or portions of the matrix are consolidated and / or hardened (bio-binding). More specifically, assume that the consolidation and / or hardening represents an operation in which a (living) organism, its portion, or an enzyme preferably obtained from or produced therefrom is used to form carbonates or to induce and / or catalyze the formation of carbonates. The formed carbonates bind portions of the matrix, or portions of the matrix to be reduced from forming dust, thereby consolidating and / or hardening the matrix or portions thereof. Therefore, in the context of this document, the formed carbonates represent the component with the lowest content of the bio-binding agent.

[0016] Furthermore, in the context of this invention, the consolidation and / or hardening are modified by water-soluble and / or water-dispersible and / or water-emulsifiable viscosity-modifying compounds, as described below.

[0017] The term "reduction of dust formation" (currently also referred to simply as "dust control" and "dust suppression") specifically refers to the sustained reduction or potential complete reduction (i.e., prevention) of dust particle generation, preferably dust generated in mining, construction, use of unpaved roads, and / or agriculture, and more preferably dust generated in spoil heaps and / or waste dumps. In this context, the term "sustainable" should be understood to mean that after one minute of exposure to wind in a wind tunnel at 12 m / s (based on the conditions described in Example 1, determined on the model matrix), the emission-related weight loss of the model matrix is ​​within the same order of magnitude for a period of at least 24 hours, preferably at least 48 hours, more preferably at least 3 days, and most preferably at least 4 days (after a single application of the mixture of the invention), i.e., the emission-related weight losses determined during this period differ from each other by less than 10 times. For example, if the initial emission-related weight loss is 0.1%, and the emission-related weight loss 24 hours after the first is 0.9%, then the emission-related weight loss is within the same order of magnitude. Conversely, if the initial emission-related weight loss is 0.1%, and the subsequent 24-hour period is 1.0% or greater, then the emission-related weight loss is not in the same order of magnitude.

[0018] Since dust suppression leads to a reduction in erosion, the present invention also relates to uses for reducing erosion.

[0019] According to the invention, this is primarily achieved through the aggregation of dust particles to form larger aggregates, also known as consolidation. The result of this aggregation is the formation of a hard shell on the matrix surface, thereby preventing the rotation of the underlying particles. However, as described below, no conclusions can be drawn, or only insufficient conclusions, from the (initially) hard shell regarding the duration of achievable dust suppression. Therefore, it is assumed that the dust suppression activity derives from other effects.

[0020] According to the present invention, the mixture comprises or consists of the following components:

[0021] (i) One or more carbonate-forming organisms and / or enzymes (i.e., organisms and / or enzymes capable of forming carbonates or inducing and / or catalyzing carbonate formation);

[0022] (ii) at least one substance used to form carbonates;

[0023] (iii) At least one water-soluble and / or water-dispersible and / or water-emulsifiable viscosity-modifying compound selected from the group consisting of:

[0024] Compounds with calcium affinity, especially those with calcium-binding functional groups selected from carboxylic acids, carboxylates, carbonyl groups, alcohols, alkoxides, thiols, thiols, sulfates, sulfonates, amines, amides, catechols, quinones, phosphates, and phosphonates; and

[0025] Compounds with carbonate affinity, especially compounds with carbonate-binding functional groups, which are composed of cations, anions and / or neutral functional groups, more preferably including cations, and more particularly monovalent and polyvalent cations, such as quaternary ammonium compounds, monovalent, divalent or trivalent metal cations, carboxylic acids, sulfonic acids, peroxycarboxylic acids, thiocarboxylic acids, sulfinic acids, sulfenic acids, amides, amines, hydrazine and thiols;

[0026] (iv) Optionally: one or more cation sources; and

[0027] (v) Optionally: one or more adjuvants.

[0028] For clarification only, it should be mentioned that mixtures according to the invention always contain different compounds and / or components (ii) and (iii). This means that one and the same substance or one and the same compound in the same mixture cannot be used simultaneously as component (ii) and component (iii).

[0029] Since bio-gluing primarily originates from the activity of enzymes and / or (living) organisms, which are typically sensitive to environmental conditions, it is not readily possible, or at least not technically relevant, to predict that bio-gluing will continue in the presence of other compounds.

[0030] However, surprisingly, the mixtures described herein have shown to produce a more durable bio-binder, thereby improving dust control. A particularly noteworthy fact is that these effects are sustained after only a few applications, or even just one application. This invention is primarily based on the understanding that the apparent initial correlation between fracture strength and dust suppression no longer exists after two days (although a correlation may be observed in individual cases). For example, treating the ground with calcium lignosulfonate creates a thin, fragile layer that, while strong, effectively no longer suppresses dust upon breakage.

[0031] Therefore, for example, ground hardness determined by fracture strength is not suitable for predicting the possible dust behavior of treated ground over a long period of time. This means that mixtures that result in less robust ground may still exhibit excellent dust suppression properties. It can be assumed that the viscosity of the deeper ground / matrix plays a greater, or even dominant, role in persistent dust suppression compared to the (initial) surface hardness, especially after the sample has been subjected to mechanical stress. For persistent dust suppression, it is important that the emission reduction effect of the sample does not diminish after mechanical exposure. In the mixtures according to the invention, the matrix viscosity is largely achieved by water-soluble and / or water-dispersible and / or water-emulsifiable viscosity-modifying compounds (hereinafter referred to as viscosity-modifying compounds), and particularly effective dust control is generally achieved through the interaction of components (i), (ii), and (iii).

[0032] Another advantage of the mixture used according to the invention is its particularly rapid hardening and / or formation of a particularly fracture-resistant bio-binder, thereby achieving further improved dust control.

[0033] In the context of component (iii), the term “water solubility” means that a compound has a solubility in water of at least 1 g / L, preferably at least 5 g / L, more preferably at least 10 g / L, more preferably at least 20 g / L, more preferably at least 50 g / L, and most preferably at least 100 g / L (all measured at 20°C).

[0034] In the context of component (iii), the terms “water-dispersible” or “water-emulsifiable” mean that a compound is dispersible or emulsifiable in water at a rate of at least 1 g / L, preferably at least 5 g / L, more preferably at least 10 g / L, more preferably at least 20 g / L, more preferably at least 50 g / L, and most preferably at least 100 g / L (all measured at 20°C).

[0035] To determine the water solubility, water dispersibility, or water emulsification of a compound, the following procedure can be used: To determine the water solubility of solid, paste-like, and gel-like compounds (e.g., polyvinyl acetate 20, polycarbonate, long-chain fatty acids, and starch), a defined amount of the compound (e.g., 5 g) is placed in a defined amount of water (e.g., 100 mL distilled water) and stirred at 20°C for 24 hours. The system is then filtered (e.g., using Homyl 80–120 μm quantitative filter paper). The filter paper is then professionally dried and weighed. The determined mass is subtracted from the mass of the filter, i.e., the mass of the residue, in grams. The difference between the defined amount of the compound (e.g., 5 g) and the mass of the residue (in grams) is divided by the defined volume of water (e.g., 0.1 L) to obtain the water solubility of the corresponding compound (in grams per liter).

[0036] To determine the water dispersibility of solid, paste, and gel substances, a defined amount of the compound (e.g., 50 g) is placed in a defined amount of water (e.g., 1000 mL of distilled water) and homogenized at 20°C (e.g., using...). Use an LC75 dissolver (at 15,000 rpm for 5 minutes). Then centrifuge the mixture (e.g., at 100 g for 2 minutes). Decant the supernatant, professionally dry the precipitate, and weigh it. The determined mass is the mass of the precipitate after centrifugation. The difference between the defined amount of the compound (e.g., 50 g) and the mass of the precipitate after centrifugation, divided by the defined amount of water (e.g., 1 L), is the water dispersibility of the substance.

[0037] To determine the water solubility or water emulsification of a liquid substance (e.g., rapeseed oil), the following procedure can be used: A defined amount of the compound (e.g., 5 g) is mixed with a defined amount of water (e.g., 100 g of distilled water), and the system is stirred for 24 hours. The mixture is then transferred to a separatory funnel. The mixture is allowed to stand in the separatory funnel for 5 minutes. If no phase separation occurs after this time, the mixture is allowed to stand for another 2 hours, preferably another 10 hours. If no phase separation occurs, the compound is considered to be water-soluble. In this example, the water solubility of the compound is at least 50 g / L. If phase separation does occur, the phases are separated in the separatory funnel, and the organic phase is dried with sodium sulfate. The weight of the dried organic phase is determined (mass of the organic phase in grams). The water emulsification of the liquid compound is obtained by dividing the difference between the defined amount of the compound (e.g., 5 g) and the mass of the organic phase (in grams) by the defined amount of water (e.g., 0.1 L).

[0038] A further preferred separation technique for the dispersed and undispersed fractions is centrifugation. After appropriate drying, the mass of the residue (in grams) can be determined, and the water solubility or water dispersibility can also be determined therefrom.

[0039] To achieve the desired or enhanced dispersibility, in the context of this invention, it may and / or is advantageous to add surfactants, such as emulsifiers and / or dispersants and / or stabilizers, to the mixture. This procedure may also achieve the effects described herein, preferably synergistic effects related to consolidation, even for viscosity-modifying compounds with relatively poor dispersibility.

[0040] In the context of component (iii), the term "viscosity modification" now refers to a compound present in a mixture in a minimum amount sufficient to modify the viscosity properties of the model matrix described herein (washed and dried silica sand graded 0-2 mm; see Example 1). This means that compounds that fundamentally (in sufficiently high amounts) possess viscosity-modifying properties but are present in the mixture in amounts below the minimum are not considered component (iii). The term "modification" now specifically refers to an increase in the viscosity forces between matrix particles. An indicator of viscosity-modifying properties is the presence of at least one chemical group capable of binding calcium and / or carbonates. Therefore, a preferred viscosity-modifying compound has calcium affinity and / or carbonate affinity.

[0041] The water-soluble and / or water-dispersible and / or water-emulsifiable viscosity-modifying compounds specifically described herein are the defined compounds of component (iii) in the sense of this invention. The compounds specifically described in each case represent preferred embodiments.

[0042] Whether another compound (the test compound) is an ingredient (iii) in the sense of this invention can be determined by measuring the emission-related weight loss of the model matrix after exposure to wind at a wind speed of 12 m / s for one minute at a predefined time following a single application of the mixture, which includes the test compound as well as ingredients (i) and (ii) (test values), and comparing it with the emission-related weight loss (comparison value) that occurred at a predefined time following a single application of a corresponding mixture excluding the compound under study. Those skilled in the art will recognize that the effect may depend on the amount used. Therefore, such comparisons can be made for different amounts of the test compound. If the test value is lower than the comparison value, the tested amount of the test compound is a viscosity-modifying compound in the sense of this invention. Whether a compound meets the desired solubility distribution can be determined by a simple solubility test (as known to those skilled in the art and also described herein).

[0043] The predefined time can be 24, 36, 48, 60, and / or 72 hours after application. Those skilled in the art will recognize that moisture content affects dust suppression. The inventors observed that the synergistic effect is particularly strong if the substrate is completely dry (approximately 4 days later). Therefore, it is preferable to conduct tests when the substrate is completely dry, i.e., for example, 3, 3.5, 4, 4.5, or 5 days after application.

[0044] Whether a compound (test compound) is calcium-affinity-binding can be determined by calcium affinity chromatography. This method can be traced back to Porath J et al. (Porath J et al., Nature. 1975. 258(5536): 598-599). Specifically, a protocol based on Campbell's Journal of Biochemistry. 1991. 19(4): 387S: agarose gel 6FF (GE Healthcare, Life Sciences Division) can be used, with the gel layered in a column (9 x 100 mm) and washed four times with a calcium chloride solution (5 mg / mL). The wash solution consists of a volume corresponding to a single column volume (as defined herein). The gel is washed once with Tris acetate buffer (pH 8.2, 0.1 M) containing sodium chloride (0.1 M) to remove unbound calcium ions. The test compound is dissolved, emulsified, or dispersed in Tris acetate / sodium chloride buffer at a concentration of 1 g test substance per liter. If the physicochemical properties of the test substance make it insoluble in the appropriate buffer, the technician will select a suitable buffer. The detection method used is absorption spectroscopy at a wavelength of 280 nm. The extinction coefficient of the test compound should be professionally determined beforehand and should exceed 1000 L mol. -1 cm -1 Or 40L g -1 cm -1 Otherwise, those skilled in the art will select a suitable wavelength with an extinction coefficient not exceeding the above values. If this is not possible at any wavelength, the elution amount should be determined by gravimetric analysis and / or atomic spectroscopy. The test substance will be applied to the column. In this case, the amount of test substance used is determined by the volume applied and the concentration of the test substance, which is also the amount of test substance used (as defined herein). After loading the test substance, the column is washed with Tris acetic acid / sodium chloride buffer and sampling begins. The column is washed three times with Tris acetic acid / sodium chloride buffer, and the fractions are collected. The mass of the test compound in the eluent is determined by absorption spectroscopy and summed. The sum of the masses of the test compounds in each fraction is the eluted test compound (as defined herein). The eluted test compound is divided by the amount of test substance used. If the ratio of these compounds is less than 0.98, the test compound exhibits calcium affinity. More preferably, the column can be washed four times with ethylenediaminetetraacetic acid (EDTA) solution (10 mM) to elute the calcium-affinity test compound from the column. The amounts of the test compound in the eluent will be determined by absorption spectroscopy and summed. This sum is the EDTA-eluted test compound (as defined herein). If the ratio of the EDTA-eluted test compound to the amount of test substance used is greater than 0.02, the compound is a calcium-affinity compound.

[0045] Whether a compound (test compound) is carbonate-affinity can be determined by the following assay: the test compound is dissolved, emulsified, or dispersed in distilled water at a concentration of 1 g / L; this solution is component A. A 200 g / L sodium carbonate solution is prepared as component B. 1 mL of completely homogenized component A is added to 10 mL of component B with stirring, and the mixture is incubated for 48 hours. If precipitation occurs and / or a large amount of gas is released, the compound is carbonate-affinity (qualitative detection). This determination can also be performed (semi-)quantitatively by the Scheibler carbonate assay or by developments of the aforementioned techniques (e.g., as described in Horváth, B. et al., A Simple Method for Measuring Soil Carbonate Content, Journal of the American Society for Soil Science 2005, 69, 1066-1068). For this purpose, it is necessary to determine the volume of gas formed when the test substance is mixed. The resulting precipitate is separated from the solution by centrifugation and dried. Subsequently, the dried precipitate is mixed with acid, and the resulting gas volume is measured. The compound in question has carbonate affinity if the sum of the gas volumes after the two measurements is greater than 0.1 mL of gas / 1 g of test substance used. Further preferred are mixtures characterized by a dust-suppressing effect (also referred to herein as a dust reduction effect) produced by components (i), (ii), and (iii) greater than the sum of the dust-suppressing effects produced by components (i) and (ii) and the dust-suppressing effect produced by component (iii). In other words, the preferred mixture is a synergistic mixture that ensures a particularly effective dust-suppressing effect over a long period.

[0046] The dust suppression effect can be determined by measuring the emission-related weight loss of the model matrix after one minute of exposure to wind at 12 m / s (based on the conditions described in Example 1, determined on the model matrix) at a defined time (e.g., 24 hours, 48 ​​hours, 3 days, 4 days, etc.) following a single application of the corresponding component.

[0047] The preferred mixture includes the following substances as component (ii):

[0048] Urea and its salts; organic acids such as lactic acid and its salts, preferably carboxylates and their esters; gluconic acid and its salts, preferably carboxylates and their esters; acetic acid and its salts, preferably carboxylates and their esters; formic acid and its salts, preferably carboxylates and their esters; propionic acid and its salts, preferably carboxylates and their esters; butyric acid and its salts, preferably carboxylates and their esters; valeric acid and its salts, preferably carboxylates and their esters; formic acid and its salts, preferably carboxylates and their esters; maleic acid and its salts, preferably carboxylates and their esters; succinic acid and its salts, preferably carboxylates and their esters; pyruvic acid and its salts, preferably carboxylates and their esters; acetoacetic acid and its salts, preferably carboxylates and their esters; acetopropionic acid and its salts, preferably carboxylates and their esters; oxaloacetic acid and its salts, preferably carboxylates and their esters; citric acid and its salts, preferably carboxylates and their esters; fruit acids, preferably malic acid and its salts, preferably carboxylates and their esters; citric acid and its salts, preferably carboxylates and their esters; fumaric acid and its salts, preferably carboxylates and their esters; gluconic acid and its salts; Salts, preferably carboxylates and esters; glycolic acid and its salts, preferably carboxylates and esters; mandelic acid and its salts, preferably carboxylates and esters; oxalic acid and its salts, preferably carboxylates and esters; salicylic acid and its salts, preferably carboxylates and esters; α-hydroxyoctanoic acid and its salts, preferably carboxylates and esters; and tartaric acid and its salts, preferably carboxylates and esters; peptides, preferably containing non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine, and / or glutamic acid; amino acids, preferably non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine, and glutamic acid, preferably carboxylates and esters; plant and animal complex matrices, especially peptones, yeast extracts, meat extracts, nutrient broths, and casein amino acids; industrial residue streams, especially corn extracts, lactose mother liquors, and protein lysates, preferably derived from peas, meat, potatoes, or tomatoes; and anaerobic substances, preferably carbon dioxide and methane.

[0049] The particularly preferred mixture includes, as component (ii), urea, acetate, formate, lactate, propionate, pyruvate, glucose, sucrose, fructose, glycerol, gluconate, lactose and / or amino acids.

[0050] The preferred mixture includes the following compounds as component (iii):

[0051] Lignosulfonates, especially calcium lignosulfonate, calcium formate, calcium propionate, calcium lactate, calcium acetate, calcium pyruvate, calcium salicylate, caseinate, albumin, alanine, asparagine, glycine, glutamine, glutamic acid, lysine, non-proteinogenic amino acids, yeast extract, albumin, polyvinyl alcohol, starch ether, magnesium sulfate, humic acid, alkali metal silicates, styrene-acrylate dispersions, polyvinyl acetate dispersions, polyacrylonitrile dispersions, polyacrylic acid, polyacrylamide, ethylene-vinyl acetate dispersions, and styrene-butadiene dispersions.

[0052] Any references to (polymer) dispersions in this article also include the polymers discussed.

[0053] In a further preferred mixture, components (ii) and (iii) (and optional components) consist of one of the following combinations:

[0054]

[0055]

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[0059]

[0060]

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[0065]

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[0070]

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[0074]

[0075] Unless otherwise stated, the substances (acids) mentioned herein may also be in the form of salts, preferably carboxylates or esters.

[0076] Those skilled in the art will recognize that the amount of component (iii) used depends largely on its own (e.g., physicochemical) properties and the properties of other components of the mixture used according to the invention, as well as the properties of the matrix, and will accordingly select a suitable combination and amount of component (iii). When the compound of component (iii) is present in a culture medium in which component (i) is used as a bacterial culture, its amount is generally too low to achieve the desired effect. The amount introduced by the bacterial culture is generally so low that the compound is present only in trace amounts in the resulting mixture.

[0077] In the preferred mixture, component (iii) is present in an amount of at least 0.5 wt%, preferably at least 1.0 wt%, more preferably at least 1.5 wt%, more preferably at least 2.0 wt%, more preferably at least 2.5 wt%, and most preferably at least 3.0 wt% (based on the total mass of components (i), (ii), and (iii) in each case). In the preferred mixture, component (iii) is present in an amount of at most 85 wt%, preferably at most 75 wt%, more preferably at most 65 wt%, more preferably at most 55 wt%, more preferably at most 45 wt%, and most preferably at most 35 wt% (based on the total mass of components (i), (ii), and (iii) in each case). In particularly preferred mixtures, component (iii) is present in an amount of 0.5 wt% to 85 wt%, preferably 1.0 wt% to 75 wt%, more preferably 1.5 wt% to 65 wt%, more preferably 2.0 wt% to 55 wt%, more preferably 2.5 wt% to 45 wt%, and most preferably 3.0 wt% to 30 wt% (in each case based on the total mass of components (i), (ii) and (iii)).

[0078] Further preferred is a mixture of components (iii) selected from the group consisting of the following substances:

[0079] (iii-1) (Bio) polymers, selected from the group consisting of:

[0080] Cellulose and its derivatives, starch and its derivatives, lignin and its derivatives, especially lignin sulfonate and sulfate lignin, pectin and its derivatives, humic acid and its derivatives;

[0081] Chitin and its derivatives, chitosan and its derivatives, cyclodextrin and its derivatives, dextrin and its derivatives,

[0082] Natural adhesives, hydrogel forming agents, cold-soluble and / or hot-soluble (plant) gums, latex, rubber and their derivatives;

[0083] Protein sources and / or peptides containing at least one of the following amino acids: alanine, glycine, lysine, asparagine, glutamine, glutamic acid, non-proteinogenic amino acids; starch ethers and starch esters, yeast and its derivatives and extracts;

[0084] Liquid and dry polymer dispersions or polymers, comprising: acids, especially acid anhydrides, sulfonic acids, sulfinic acids, hyposulfonic acids, carboxylic acids, peroxycarboxylic acids and thiocarboxylic acids and their salts, sulfoxides, cyanates, thiocyanates, esters, ethers, thioethers, ethylene oxide, cyclothioethane, amines, imines, hydrazines, hydrazones, amides, sulfates, nitriles, aldehydes, thioaldehydes, ketones, thioketones, oximes, alcohols, thiols, free radicals, halogens, silanes, siloxanes, phosphates, phosphonates, alkyl, allyl and aryl groups and their derivatives.

[0085] (iii-2) (Poly)saccharides and extracellular polymeric substances (EPS), and their derivatives in each case, selected from the group consisting of microbial extracellular polysaccharides, preferably including: lactose, sucrose, glucose, glucosamine, mannose, glycerol, acetate, gluconate, fructose, inulin and combinations thereof or consisting of thereof;

[0086] (iii-3) Carboxylic acids, selected from formic acid, maleic acid, succinic acid, butyric acid, propionic acid, acetic acid, pyruvic acid, acetoacetic acid, levulinic acid, oxalic acid, citric acid, and fruit acids, preferably malic acid, citric acid, fumaric acid, gluconic acid, glycolic acid, mandelic acid, oxalic acid, salicylic acid, α-hydroxyoctanoic acid, and tartaric acid; fatty acids, preferably short-chain and medium-chain fatty acids; and lactic acid and its salts (in each case), preferably the group consisting of carboxylates and their esters.

[0087] (iii-4) Inorganic binders, minerals and salts selected from cementing agents, including their derivatives, preferably CEM I, CEM I, CEM III, CEM IV, CEM V, CEM VI, alumina cementing agents, magnesium oxide cementing agents, phosphate cementing agents, gypsum, sodium silicate, potassium silicate and lithium silicate, and other water glass derivatives, calcium carbonate and its derivatives, alumina, aluminum hydroxide, calcium sulfate, calcium hydroxide, calcium oxide, magnesium sulfate, microsilica, kaolin, bentonite and (hydrated) lime;

[0088] (iii-5) Amino acids selected from alanine, glycine, lysine, asparagine, glutamine, glutamic acid, non-proteinogenic amino acids and their salts (in each case), preferably the group consisting of carboxylates and their esters and amides.

[0089] As defined herein, the term "polymer" includes all water-soluble and / or water-dispersible and / or water-emulsifiable (as defined herein) polymers. Polymer dispersions constitute a subgroup of polymers.

[0090] In some embodiments, the liquid or dry polymer dispersion or the liquid or dry polymer is a polymer or copolymer containing one of the following monomers or composed of two or more different monomers from the following: ethylene, propylene, butadiene, butene, styrene, isoprene (and other allyl and acrylic monomers), acrylic acid and its salts, preferably carboxylates and their esters, vinyl monomers, such as vinyl acetate, vinyl chloride, vinyldecanoate, vinylpyrrolidone and vinylimidazolium and their respective derivatives, isocyanates and their salts, more preferably cyanates, especially mono, di and polyisocyanates, alcohols, preferably polyols, more preferably diols, triols and tetraols, amines, preferably polyfunctional... Aminoamines, more preferably diamines, triamines, tetraamines, especially diaminobenzene, ethylenediamine and diethylenetriamine, epichlorohydrin, bisphenols, preferably bisphenol A and bisphenol F, 2-ethyl-2-oxazoline, ethylene oxide, propylene oxide, urea, melamine, phenol, formaldehyde, siloxanes, tetramethylsilane, trimethylchlorosilane, dimethyldichlorosilane, methyltrichlorosilane, tetrachlorosilane, acrylonitrile, maleic acid, hydroxy acids, preferably hydroxy fatty acids, dicarboxylic acids, preferably oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, succinic acid, azelaic acid, sebacic acid and terephthalic acid, acrylamide, amino acids, non-proteinogenic amino acids, monosaccharides, disaccharides, oligosaccharides and their derivatives.

[0091] In a further preferred mixture, component (iii) is selected from the group consisting of the following substances:

[0092] Calcium lignosulfonate, sodium lignosulfonate, potassium lignosulfonate, magnesium lignosulfonate, ammonium lignosulfonate, sulfate lignin, humic acid and its salts, preferably carboxylates and their derivatives.

[0093] Fibers and fibrous materials, selected from the group consisting of cellulose fibers, wood fibers, and wood cellulose fibers.

[0094] gum arabic, xanthan gum, alginate, and agar.

[0095] Protein sources and / or amino acids are selected from the group consisting of casein, albumin, yeast extract, peptone, caseinate, calcium caseinate, milk powder, alanine, glycine, lysine, asparagine, glutamine, glutamic acid, and non-proteinogenic amino acids. Residual substances and industrial substances are selected from the group consisting of corn leachate, lactose mother liquor, protein lysate, molasses, and protein waste, preferably derived from yeast, meat, fruit and vegetable production, dairy industry, and papermaking.

[0096] Liquid and dry polymer dispersions or polymers selected from the group consisting of polyhydroxybutyrate, polylactide, polybutylene succinate, polyacrylic acid, polymethacrylate, poly(2-hydroxyethyl methacrylate), polyvinyl alcohol, polyvinyl acetate, polyvinylpyrrolidone, poly(2-ethyl-2-oxazoline), polystyrene, polyamide, styrene-butadiene, styrene-acrylate, styrene-acrylate, acrylic acid, vinyl acetate, isocyanates, epoxides, and polyamino acids. In particularly preferred mixtures, component (iii) is selected from the group consisting of lignin sulfonates, especially calcium lignin sulfonate, yeast extract, albumin, starch ether, alanine, lysine, styrene-acrylate dispersions, polyvinyl alcohol, polyvinyl acetate dispersions, styrene-butadiene dispersions, humic acid, alkali metal silicates, and combinations thereof.

[0097] Further preferred is a mixture of components (ii) selected from the group consisting of the following substances:

[0098] Urea and its salts; organic acids such as lactic acid and its salts, preferably carboxylates and their esters; gluconic acid and its salts, preferably carboxylates and their esters; acetic acid and its salts, preferably carboxylates and their esters; formic acid and its salts, preferably carboxylates and their esters; propionic acid and its salts, preferably carboxylates and their esters; butyric acid and its salts, preferably carboxylates and their esters; valeric acid and its salts, preferably carboxylates and their esters; peptides, preferably containing non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine and / or glutamic acid; amino acids, preferably non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine and glutamic acid and their salts, preferably carboxylates and their esters; plant and animal composite matrices, especially peptones, yeast extracts, meat extracts, nutrient broths and casein amino acids; industrial residual substrates, especially corn extract, lactose mother liquor, protein lysates, preferably derived from peas, meat, potatoes or tomatoes; anaerobic matter, preferably carbon dioxide and methane.

[0099] The condition is that components (ii) and (iii) are different from each other.

[0100] The preferred use is as described above, wherein the mixture is present in liquid form, as a gel, paste, or powder.

[0101] The mixtures used according to the present invention may be in the form of liquid, gel, paste or powder mixture, or in the form of two, three, four or more separate liquid and / or gel and / or paste and / or powder premixes (which are mixed with each other before or during use).

[0102] In particular, mixtures or premixes in powder form have a particularly long storage stability, preferably at least 12 to 24 months.

[0103] The powder form of the mixture or premix can be obtained by standard methods familiar to those skilled in the art, such as spray drying, freeze drying, (low temperature) vacuum drying, fluidized bed drying and / or filtration with the aid of a filter aid.

[0104] In this context, "powder" means that, based on the total weight of the premix or mixture used according to the invention, the amount of liquid component (preferably water) in the mixture is 10 wt% or less, preferably 5 wt% or less, preferably 2.5 wt% or less, more preferably 1.0 wt% or less, and most preferably 0.1 wt% or less.

[0105] The amount of liquid component (preferably water) in a mixture or premix can be determined using standard methods known to those skilled in the art. For example, a gravimetric analysis of the liquid component can be performed, in which a sample is weighed, then heated for a sufficient time at a temperature above the boiling point of the liquid component until dry, and then weighed again. Based on the weight difference before and after drying, the amount of liquid component (preferably water) can be determined (in wt%).

[0106] According to another embodiment, the mixture used according to the invention may be in the form of a gel or paste mixture, or may be present and / or used in the form of two, three, four or more separate solid and / or liquid and / or gel and / or paste premixes (which are mixed with each other before or during use).

[0107] The use of the mixture according to the invention advantageously results in a bio-binding agent layer thickness particularly suitable for the purposes described herein. Preferably, in this case, a bio-binding agent layer with a thickness of at least 1 mm, preferably at least 3 mm, more preferably at least 10 mm is obtained. Further preferably, if the layer thickness is not greater than 100 mm, preferably not greater than 50 mm, more preferably not greater than 35 mm, more preferably not greater than 30 mm, it is particularly preferred that the thickness of the integrally formed bio-binding agent layer is in the range of 1 mm to 100 mm, preferably 10 mm to 50 mm, more preferably 10 mm to 35 mm, more preferably 10 to 30 mm. The thickness of the bio-binding agent layer covers the matrix area that is solidified by adding the mixture. The thickness of the bio-binding agent layer can be determined manually by calipers after the layer is mechanically broken. Alternatively, depending on the solidified thickness, it can be determined using various (non-destructive) measurement methods (e.g., manual instruments such as MIT-SCAN-T2) in the fields of construction, agriculture, geology, or other applications.

[0108] According to another preferred embodiment, the use of the mixture according to the invention, as described herein, results in a water-permeable biocementing layer, i.e., permeable or semi-permeable. This is particularly advantageous because, for example, external rainwater can also permeate into the formed biocementing layer without obstruction and flow out. The permeability of a sample is typically reported as the flow rate of water through the sample over a defined time period. It can be expressed as permeability (in cm / h, mm / h, or cm / day) or as a permeability coefficient (in m / s). The statement of the permeability coefficient allows the sample, preferably a soil sample, to be classified, for example, as permeable, semi-permeable, and impermeable.

[0109] In the context of this article, the term "permeable bio-cement layer" refers to a (water) permeability coefficient greater than 10. -5 Up to 10 0 A bio-cementing layer with a permeability of m / s, the term "semi-permeable bio-cementing layer" indicates a (water) permeability coefficient greater than 10. -9 Up to 10 -5 A bio-cement layer with a permeability of m / s, and the term "impermeable bio-cement layer" indicates a (water) permeability coefficient of 10. -11 (or smaller) up to 10 -9 A bio-cementing layer of m / s. Commonly used methods for determining permeability include laboratory methods (e.g., core probe and subsequent determination of water-saturated permeability in the laboratory) and field methods (e.g., determination of permeability using a dual-ring permeameter).

[0110] One preferred embodiment relates to the use of a mixture as defined herein, wherein the (water) permeability coefficient of the formed bio-binding agent layer is greater than 10. -9 Up to 10 0 m / s, preferably greater than 10 -9 Up to 10 -3 m / s, more preferably greater than 10 m / s -8 Up to 10 -3 m / s.

[0111] The use according to the invention demonstrates strong functionality under real (environmental) conditions, ease of use (typically through a single application), and reduction or avoidance of toxic substances. Furthermore, it can be combined with other dust suppression measures. In some cases, it may be necessary to reverse the consolidation. An advantage of the use according to the invention is its reversibility, meaning that the biocementation of the matrix or a portion of the matrix can be reversed when needed, for example, by applying a suitable acid or by mechanical disruption. The matrix or a portion of the matrix can be obtained in this way, for example, for the growth of crops. Therefore, one embodiment relates to the use of mixtures as defined herein, wherein the biocementation of the matrix or a portion of the matrix can be reversed or preferably reversed.

[0112] Preferred is the use as described above, wherein one or more, or all, organisms are selected from the group consisting of microorganisms, preferably from the phylum Firmicutes, preferably the class Bacilli, preferably the order Bacillales, preferably the family Planococcaceae or Bacillaceae, preferably from the group consisting of microorganisms of the genera Sporosarcina, Lysinibacillus, or Bacillus, preferably from the group consisting of Sporosarcina pasteurii, Sporosarcina ureae, Lysinibacillus sphaericus, Lysinibacillus fusiformis, Bacillus megaterium, and Lysinibacillus. sp.), Bacillus pseudofirmus, Bacillus halodurans, or Bacillus cohnii;The phylum Proteobacteria, preferably alpha-proteobacteria, gamma-proteobacteria, delta-proteobacteria, or eppsilon-proteobacteria, preferably order Enterobacteriales, Myxococcales, Campylobacterales, Pseudomonadales, or Calobacterales, preferably family Enterobacteriaceae. The group consists of microorganisms from the families Myxococcus, Helicobacter, Pseudomonadaceae, or Caulobacteraceae, preferably Proteus, Myxococcus, Helicobacter, Pseudomonas, or Brevundimonas, and more preferably Proteus vulgaris or Proteus mirabilis. The group consisting of microorganisms including *Myxococcus xanthus*, *Helicobacter pylori*, *Pseudomonas aeruginosa*, or *Brevundimonas diminuta*; and *Actinobacteria*, preferably *Actinobacteria*, preferably *Actinomycetales*, preferably *Brevibacteriaceae* or *Micrococcineae*, preferably *Brevibacterium* or *Micrococcaceae*, and preferably selected from *Brevibacterium linens* or *Arthrobacter crystallopoietes*.The group consisting of microorganisms from the phylum Cyanobacteria, preferably the class Cyanobacteria, preferably the order Synechococcales, preferably the family Synechococcaceae, preferably the genus Synechococcus, and preferably the species Synechococcus; as well as aerobic bacteria, anaerobic bacteria, facultative anaerobic bacteria, and their intermediate stages.

[0113] This also includes all variants, serotypes, mutants, and spores, as well as any derived genetically modified microorganisms.

[0114] The aforementioned organisms, preferably microorganisms, may exist (together or separately) in a liquid, such as a buffer solution, solvent, nutrient medium and / or mixtures thereof, and these mixtures may also be lyophilized mixtures or may exist in powder form.

[0115] According to the present invention, the organism capable of forming carbonates or inducing and / or catalyzing carbonate formation is part of the mixture used.

[0116] Alternatively, it is conceivable, and also provided in the context of this invention, that (native) organisms present in and / or separated from the substrate (preferably soil), cultured in a laboratory, and then introduced onto / into the substrate are capable of forming carbonates or inducing and / or catalyzing carbonate formation. In this case, alternative or equivalent embodiments are conceivable (and are accordingly included herein as according to the invention), wherein the organisms of the mixture used according to the invention and the (native) organisms in / on the substrate together form carbonates or induce and / or catalyze carbonate formation, and / or the mixture used herein does not itself contain organisms capable of forming carbonates or inducing and / or catalyzing carbonate formation. According to a preferred embodiment, the components (i) of the mixture used according to the invention comprise, or consist of, organisms capable of forming carbonates or inducing and / or catalyzing carbonate formation, or combinations of two or more organisms with organisms that do not have this capability.

[0117] According to a preferred embodiment, the components (i) of the mixture used according to the invention comprise, or consist of, a combination of aerobic bacteria, anaerobic bacteria and / or facultative anaerobic bacteria and / or intermediate stages thereof.

[0118] According to another preferred embodiment, the components (i) of the mixture used according to the invention include organisms capable of decomposing urea to form carbonates or urea decomposition inducing and / or catalyzing carbonate formation, or combinations of two or more organisms and organisms that cannot decompose urea and / or cannot form carbonates or induce and / or catalyze carbonate formation, or are composed of the like.

[0119] In this context, those skilled in the art will recognize that the bio-binding (as defined herein) of mixtures used according to the invention is particularly efficient within the defined cell count spectrum of the organisms used. Based on internal studies, the cell count of the organisms in the mixtures used according to the invention is preferably at least 10. 7 cells / mL, more preferably at least 10 8 cells / mL, and / or preferably up to 10 12 cells / mL, more preferably up to 10 10 cells / mL and more preferably up to 10 9 Cells / mL. According to a preferred embodiment, the cell count of the organism in the mixture used according to the invention is 10-1 cells / mL. 8 Up to 10 9 Cells / mL.

[0120] Preferred is the use described above, wherein one, two or more, or all of the enzymes are selected from the group consisting of urease, asparaginase, carbonic anhydrase, and metabolic enzymes.

[0121] In the case of metabolic enzymes, the enzymes discussed in the context of this document are metabolic enzymes of one or more (micro)organisms described herein, which are capable of forming carbonates or inducing and / or catalyzing carbonate formation through, for example, the conversion of acetate and / or lactate. In the components (i) of the mixtures used according to the invention, preferably one, two, or more organisms (as defined above) capable of producing one or more of the aforementioned enzymes are used, and / or preferably, the enzymes are obtained or released from the aforementioned organisms.

[0122] When the organisms used include pathogenic organisms, in the context of this document, preferably only non-pathogenic enzymes obtained from or released from said organisms are used in component (i) of the mixture used according to the invention.

[0123] According to another preferred embodiment, in component (i) of the mixture used according to the invention, a combination of enzymes obtained or released from the aforementioned organisms and enzymes of non-microbial origin (e.g., plant enzymes) may be used. For example, urease can be obtained from soybeans and used according to the invention.

[0124] According to another preferred embodiment, in the components (i) of the mixture used according to the invention, a combination of one or more of the organisms described above capable of forming carbonates or inducing and / or catalyzing carbonate formation and one or more of the enzymes described above capable of forming carbonates or inducing and / or catalyzing carbonate formation may be used.

[0125] Carbonates can be produced via various metabolic processes through the enzymes mentioned above. For example, aerobic metabolism of organic carbon sources may occur, such as ammoniation (e.g., asparaginase), or heterotrophic metabolism may occur with organic carbon sources (e.g., calcium lactate or calcium acetate). Both processes produce carbonates. Aerobic and anaerobic photosynthesis can also be used to form carbonates, such as anaerobic denitrification, anaerobic sulfate reduction, and (anaerobic) aerobic methane oxidation.

[0126] Therefore, the bio-binding of the mixture used according to the present invention can be based on one or more of the above-described metabolic processes.

[0127] In the context of this invention, whether an organism or enzyme other than the organism or enzyme specifically described herein represents a component (i) can be determined by means of the following determination A.

[0128] (Measurement A)

[0129] (i) Providing and contacting the organism used for testing or a mixture of the organism used for testing (test organism) and component (ii),

[0130] (ii) Provide an apparatus for establishing urea decomposition and / or carbonate formation.

[0131] (iii) Combining the mixture obtained in step (i) with the apparatus in step (ii), and

[0132] (iv) Based on the apparatus of step (ii), determine whether urea decomposition and / or carbonate formation are present.

[0133] If urea decomposition and / or carbonate formation are established in step (iv), then the test organism is an ingredient (i) in the sense of the present invention.

[0134] According to a slightly modified determination A', step (i) further includes a model matrix (as described herein), and step (iv) includes determining whether a biocluster has been established, and if a biocluster has been established, then the test organism is a component (i) in the sense of the present invention.

[0135] If the component to be tested is an enzyme or a mixture of enzymes, a similar process can be performed.

[0136] The following observations can be used to select organisms in contexts suitable for use according to the present invention.

[0137] In this document (including based on selected and preferred embodiments), the component (ii) to be provided in step (i) of determination A as defined herein is defined more precisely.

[0138] In step (i) of determination A as defined herein, for example, a pure culture of the organism to be characterized (e.g., from a type set) may be provided, and / or the organism to be characterized or a mixture of organisms to be characterized may be isolated from a suitable sample (e.g., a soil sample) via a nutrient medium (e.g., Christensen urea agar, B4 medium, or M-3P medium) and cultured to form a cell culture suitable for further study. The nutrient medium used for isolation and culture may be liquid or solid. Those skilled in the art will recognize that the nutrient medium may vary depending on the requirements of the organism. Preferably, the organism is cultured to 1 x 10⁻⁶. 7 Up to 1x10 12 A cell density of cells / mL. Those skilled in the art will recognize that, for example, the culture temperature and culture medium composition are selected according to the requirements of the organism or mixture of organisms. The provided or produced cell culture is then contacted with component (ii) and an optional model matrix to form a mixture, which is then combined with the apparatus of step (ii) in step (iii).

[0139] The apparatus defined herein for establishing urea decomposition and / or carbonate formation in step (ii) of determination A is, for example, a pH indicator, an apparatus for measuring urease activity and / or one or more substances, an apparatus for measuring the amount of carbonate formed by bioaggregation and / or one or more substances, or an apparatus for measuring the degree of consolidation of the matrix (as a result of bioaggregation).

[0140] In step (iv) of determination A as defined herein, particularly the establishment of biocollagen, the presence of urea decomposition and / or carbonate formation can be qualitatively or preferably quantitatively determined.

[0141] A preferred method for the determination is, for example, to add a suitable pH indicator (e.g., phenol red, preferably at a concentration of 15 mg / L) to the mixture obtained in step (i). When urea decomposition and / or carbonate formation are present, the pH of the mixture increases, causing a portion of the indicator to change color (e.g., turning pink in the case of phenol red).

[0142] If a cation source, preferably a calcium source, is added to the mixture of step (i), and if urea decomposition and / or carbonate formation are present, a lime crust typically forms around the colony and / or on the colony of the organism in solid media. In the case of liquid nutrient media, if a cation source, preferably a calcium source (e.g., CaCl2), is readily available, and if a carbonate source (e.g., urea) is readily available, lime precipitation is typically present. Such lime crust formation or lime precipitation can also serve as visual evidence of urea decomposition and / or carbonate formation, and / or the aforementioned instances of lime crust formation or lime precipitation can be analyzed by qualitative and / or quantitative carbonate determination, preferably by means of the Scheibler method or a further development of that method (e.g., as described in Horváth, B. et al., A Simple Method for Measuring Soil Carbonate Content, Journal of the American Society for Soil Science 2005, 69, 1066-1068) for (semi-)quantitative carbonate determination.

[0143] Another method for the determination is, for example, measuring the urease activity of an organism or mixture of organisms. In this case, the organism or mixture of organisms to be analyzed is mixed with a reagent comprising buffered urea (e.g., 1.5 M urea in 0.1 M TrisHCl, pH 7.5), and the formation of the resulting ammonium ions is measured by conductivity measurement as the rise of the measurement signal over time, and the urease activity is calculated (as described, for example, in VSWhiffin, Microbial CaCO3 Precipitation for Biocement Production, Doctoral Dissertation, 2004, Murdoch University, Western Australia). The urease activity is preferably 1 x 10⁻⁶. -7 Up to 1x10 - 11 mM hydrolyzed urea min / cm / cell / mL, more preferably 1x10 -8 Up to 1x10 -10 mM hydrolyzed urea / min / cm / cell / mL, more preferably 1x10 -8 Up to 1x10 -9mM hydrolyzed urea / min / cm / cell / mL. The former approximately corresponds to a urea hydrolysis rate of 0–300 mM hydrolyzed urea / min, depending on the cell count used. Another preferred method for the determination is, for example, to measure the amount of carbonate formed by biogel by a (semi-)quantitative carbonate assay using the Scheibler method. The mixture in the study is preferably incubated in an open environment at room temperature (25°C) for 48 hours. This yields precipitated particles to be obtained by centrifugation and drying for further use. The dried particles can be used for (semi-)quantitative detection of the calcium carbonate formed, preferably by the Scheibler carbonate assay. Optionally, the dried precipitate can be weighed in advance, and the precipitation efficiency can be calculated. Optionally, concurrently, an additional qualitative determination can be performed to determine the presence of urea decomposition and / or carbonate formation. For this purpose, phenol red (15 mg / L) can be added to the mixture from step (i). If urea decomposition and / or carbonate formation are present, the supernatant discarded during particle recovery is typically pink in this case.

[0144] Another preferred method for the determination is, for example, measuring the degree of consolidation of the matrix (by carbonates formed during the biocementing process). A suitable matrix for this purpose is silica sand, preferably with a particle size of 0 to 2 mm (as a model matrix). The remaining components of the mixture in step (i) are preferably in a concentration of 5 l / m 2 (In the case of liquid mixtures) The resulting mixture is applied / introduced onto / into the matrix. Subsequent incubation should be carried out in an open system at or above room temperature for at least 2 days (preferably at least 10 days). The strength of the formed bio-binding agent layer is then determined by fracture mechanics analysis using a digital (fracture) strength measuring instrument, according to the method based on DIN EN 196-1:2005-05. A difference in fracture strength of ≥3 N (or ≥0.01 MPa), preferably ≥30 N (or ≥0.1 MPa), should be detectable compared to the control group (a control mixture without organisms applied to the matrix).

[0145] In determination A, calipers can also be used to determine the thickness of the bio-bonding agent layer; if consolidation is successful, preferably within the study range, the thickness should be an average of ≥3 mm.

[0146] According to a preferred embodiment, the components (i) of the mixture used according to the invention include organisms that, in step (iv) of determination A as defined herein, result in urea decomposition and / or carbonate formation, preferably the establishment of bioclinker, with respect to two or more of the above-described determination methods, preferably three or more, more preferably four or more, and most preferably all determination methods.

[0147] Furthermore, the preferred use is as described above, wherein component (iv) is selected from organic and inorganic calcium salts, preferably calcium nitrate, calcium acetate, calcium lactate, calcium pyruvate, calcium salicylate and calcium chloride, magnesium salts, manganese salts, zinc salts, cobalt salts, nickel salts, copper salts, lead salts, iron salts, cadmium salts, polymers, preferably cationic polymers, heavy metal cations, light metal cations, radioactive cations and mixtures thereof.

[0148] According to the invention, component (iv) may or may not be present in the mixture used according to the invention. If it is not present in the mixture, it may be present on / in the treated matrix, or may be added to / in the matrix to achieve bio-binding.

[0149] Particularly preferred is the use as described above, wherein component (iv) is present at a total concentration of 0.05 to 1 M and / or a total calcium concentration not exceeding 1.5 M.

[0150] Optionally, the mixture as described herein may include one or more adjuvants (component (v)). If component (v) is present, it is preferably selected from the group consisting of:

[0151] Natural and chemical herbicides; fungicides; molluscicides; insecticides; hydrophobic agents and wax emulsions; stabilizers; dispersants; emulsifying aids; surfactants, preferably cationic, anionic and uncharged surfactants; amines; ethanolamine; thixotropic agents; propellants; free-flowing agents, seed crystals and crystallization modifiers; complexing agents, preferably phosphonates, phosphates and polyphosphates, fatty acids; minerals and trace elements; salts, preferably halides, silicates, phosphates and sulfates; rocks, preferably pumice, sand, gravel and slate powder, rubber fragments, rubber granules and other thermoplastic elastomers, preferably derived from the tire industry; aggregates, preferably amorphous and crystalline aggregates, more preferably hydraulic, non-hydraulic and hardening materials; plant seeds, preferably monocotyledonous and dicotyledonous plants, spores, preferably bryophyte spores, plants and their parts, preferably roots, bulbs, wood and wood chips; fertilizers; bacteria capable of forming polymers; and modified biocementing substances.

[0152] Furthermore, reference is made to observations regarding the methods and mixtures of the present invention, which are correspondingly effective for applications according to the present invention. In addition, embodiments described in conjunction with dust control represent corresponding embodiments of erosion control, as well as other applications disclosed herein.

[0153] Another aspect of the present invention relates to a method for reducing dust formation and / or erosion. The method includes the following steps:

[0154] (a) Identify the substrate to be treated and reduce dust formation and / or erosion on / within the substrate.

[0155] (b) to provide mixtures or components thereof as defined herein (especially in the context of their use according to the invention),

[0156] (c) Apply the mixture or its components provided in step (b) to the substrate to be treated in an amount sufficient to achieve bio-binding, and

[0157] (d) Allows the formation of a bio-binding layer, thereby reducing dust formation and / or erosion on / in the substrate.

[0158] According to a preferred embodiment of the method of the present invention, the application is to apply the mixture or components thereof provided in step (b) onto / into the substrate to be treated. According to another preferred embodiment, the application includes application and subsequent introduction, for example by mixing the mixture or components thereof provided in step (b) onto / into the substrate to be treated. According to yet another preferred embodiment of the method of the present invention, the application is to introduce the mixture or components thereof provided in step (b) onto / into the substrate to be treated.

[0159] According to one embodiment of the method of the present invention, the matrix or a portion thereof identified in step (a) is removed from its original location and mixed with the mixture or components thereof provided in step (b) in an amount sufficient to achieve bio-binding (e.g., in a mixing device), and the resulting mixture is returned to the original location of the matrix (or to a different location where a bio-binding agent layer is to be formed), followed by step (d) as described herein. In this embodiment, step (c) of the method described herein is omitted.

[0160] Based on the form (solid / powder, liquid, gel, or paste) of the mixture or its components provided in step (b) of the method of the present invention (see the observations above), the application in step (c) can be performed in a variety of ways. For example, the powdered mixture can be dispersed onto and / or incorporated into the substrate to be treated. For example, the liquid mixture can be poured or sprayed onto the substrate to be treated and optionally subsequently incorporated into the substrate. Advantageously, generally, a single application of the mixture or its components provided in step (b) onto / into the substrate to be treated is sufficient to form a bio-binding agent layer as defined in step (d) of the method of the present invention. Preferably, a single application of the mixture provided in step (b) onto / into the substrate to be treated is sufficient to form a bio-binding agent layer as defined in step (d) of the method of the present invention.

[0161] In this document, those skilled in the art will recognize that the bio-binding in the method of the present invention (as defined herein) is carried out with a specific efficiency at a defined application volume and / or defined concentration of the mixture in step (b) (see also the preferred cell count of the organism in the mixture used according to the present invention, as described herein). Based on internal studies, the application volume of the mixture used according to the present invention (as defined above) is preferably at least 0.1 l / m³. 2 More preferably at least 0.5 l / m 2 More preferably at least 1.0 l / m 2 More preferably at least 2.0 l / m 2 At least 3.0 l / m 2 At least 4.0 l / m 2 Or at least 5.0 l / m 2 And / or preferably up to 20.0 l / m 2 More preferably up to 10.0 l / m 2 .

[0162] For the effective bio-binding process in step (d) of the method of the present invention, it is advantageous that the water content of the system consisting of the matrix (as defined herein) and the mixture used according to the present invention is greater than 10 wt% (based on the total weight of the system). If the mixture used according to the present invention is used in powder form (as defined above) in step (b) of the method of the present invention, and if the matrix in step (a) or (c) of the method of the present invention is also substantially anhydrous, and the water content of the system is 10 wt% or less (based on the total weight of the system), it is advantageous if the method of the present invention includes the additional step of adding sufficient water or an aqueous solution to the mixture or components thereof in step (b) of the method of the present invention before or after application to the matrix to be treated, such that the resulting water content of the system is greater than 10 wt% (based on the total weight of the system). And / or, a suitable amount of water or an aqueous solution may be added to the matrix to be treated before or after applying the mixture or components thereof provided in step (b) of the method of the present invention.

[0163] Furthermore, it is advantageous to avoid implementing the method during periods of heavy rainfall or strong winds when using the method of the present invention. In some cases, even before the formation of the bio-cementing layer (step (d)), heavy rainfall or strong winds can cause loss or severe dilution of the mixture used according to the present invention, which may prevent the formation of the bio-cementing layer and / or adversely affect its hardness and / or thickness. After the mixture or its components provided in step (b) of the method of the present invention are applied to / into the substrate to be treated, i.e., in step (d) of the method of the present invention, the bio-cementing layer is preferably formed during an incubation period of at least 6 hours, preferably at least 24 hours, more preferably at least 48 hours, wherein preferably there is no rain, wind, or artificial watering that would cause significant loss of the mixture used according to the present invention. In this context, it is particularly advantageous that the mixture according to the present invention, in addition to having the advantage of durable consolidation, is also capable of accelerating consolidation. Therefore, the aforementioned weathering-related losses can be largely prevented or reduced.

[0164] The incubation time required to form the bio-binding agent layer in step (d) of the method of the present invention also depends on various environmental parameters, such as room temperature or outdoor temperature and atmospheric humidity, and the application volume of the mixture. During the incubation period of at least 6 hours, preferably at least 24 hours, more preferably at least 48 hours, if rain or wind causes significant loss of the mixture or its components used according to the present invention, it is advantageous to repeat steps (b) to (d) of the method of the present invention as frequently as possible, preferably once, twice, three times or more, until the bio-binding agent layer reaches sufficient thickness and hardness. And / or, it can be proven advantageous to repeat steps (b) to (d) of the method of the present invention, preferably once, twice, three times or more, because the thickness and / or hardness of the bio-binding agent layer formed on / among the substrate due to weathering and / or natural degradation should decrease over time and thus no longer be sufficient to reduce dust formation and / or erosion.

[0165] The thickness of the bio-cementing layer can be determined manually using calipers after the layer has been mechanically fractured. Alternatively, depending on the thickness of the consolidation, various (non-destructive) measurement methods (such as the manual device MIT-SCAN-T2) can be used in construction, agriculture, geology, or other applications. The thickness of the bio-cementing layer includes the matrix area that has been consolidated due to the addition of the mixture.

[0166] The hardness of the bio-binding agent layer corresponds to its fracture strength (in Newtons (N)), which is the force that must be applied to fracture the bio-binding agent layer. Fracture of the bio-binding agent layer occurs when, upon contact with a force, the layer no longer exhibits any (plastic) deformation, resulting in the breakage of the (bio-binding) layer. Fracture is identified by measuring the decrease in force. The fracture strength (the maximum force measured) can be determined by the following method: This method is based on the standard test method for determining the strength of binders in DIN EN 196-1:2005-05. The fracture strength is measured using a digital (fracture) strength measuring instrument according to the manufacturer's instructions. The specimen is pressed into the sample (until the fracture point) using a crank-operated test bench, and the applied force is continuously measured. The average fracture strength is calculated through multiple measurements (>3). The average fracture strength is preferably from 0.5 N to 1000 N, more preferably from 3 N to 300 N.

[0167] The preferred method is the one described above, wherein the matrix is ​​selected from organic and inorganic materials, especially biological and / or anthropogenic materials, preferably metamorphic rocks, sedimentary rocks and igneous rocks and their derivatives and mixtures (in each case), and combinations thereof.

[0168] More preferably, the matrix used in the method of the present invention is selected from the group consisting of materials described in one or more of the following subgroups (Strunz H and Nickel EH, V Strunz Mineral Tables, 2001, 9th edition):

[0169] (i) Elements (including all subgroups), such as but not limited to: gold, copper, silver, zinc, tin, iron, antimony, graphite, palladium, carbon;

[0170] (ii) Sulfides and sulfonates (including all subgroups), such as but not limited to: chalcopyrite, galena, pyrite;

[0171] (iii) Halides (including all subgroups), such as but not limited to: fluorite;

[0172] (iv) Oxides and hydroxides (including all subgroups), such as but not limited to: calcium oxide, magnesium oxide, cassiterite, magnetite, hematite, ilmenite;

[0173] (v) Carbonates and nitrates (including all subgroups), such as but not limited to: calcite;

[0174] (vi) Borates (including all subgroups), such as but not limited to: borax borax, sodium borate;

[0175] (vii) Sulfates, chromates, molybdates, tungstates (including all subgroups), such as but not limited to: anhydrous potassium magnesium alum, anhydrite, magnesium sulfate, gypsum;

[0176] (viii) Phosphates, arsenates, vanadates (including all subgroups), such as but not limited to: monazite;

[0177] (ix) Silicates and germanates (including all subgroups), such as but not limited to: peridot, topaz, muscovite, talc, binders, microsilica, water glass;

[0178] (x) Organic minerals (including all subgroups).

[0179] The method includes mixtures of one or more of the above-mentioned materials, as well as substances of biological and / or anthropogenic origin and / or mixtures thereof, such as, but not limited to: soil, ash, wood, mulch, cementitious agents, calcium carbonate (including polymorphs, derivatives and mixtures, as well as natural-based (ground calcium carbonate (GCC)) and synthetic (precipitated calcium carbonate (PCC))), alumina, aluminum hydroxide, magnesium oxide, calcium oxide, calcium hydroxide and waste rock, and fine-grained residues from the processing of one or more of the above-mentioned substances or mixtures thereof (tailings); more preferably, the matrix used in the method of the present invention is selected from the group consisting of crystalline and amorphous substances and mixtures thereof.

[0180] Depending on the nature of the substrate to be treated, it can be demonstrated that mixing the substrate (or components (i), (ii), (iii) and / or (iv)) of the mixture provided in step (b) with one or more of the aforementioned additives is advantageous, in order to, for example, increase the reactivity of the substrate having the bio-cementitious agent formed in the method of the present invention. This advantageously results in a particularly hard / stable bio-cementitious agent layer, which is particularly effective in inhibiting dust formation and / or erosion.

[0181] A further preferred method is the one described above, wherein the mixture is in liquid form, present as a gel, paste, or powder (see above).

[0182] Therefore, the mixture or components of the mixture provided in step (b) of the method of the present invention may be in the form of a mixture, preferably in the form of a powder, or in the form of two, three, four or more separate liquid and / or gel and / or paste and / or powder premixes, which are mixed with each other before or during step (c) before or during application to the substrate to be treated.

[0183] Advantageously, a single implementation of steps (b) to (d) of the method of the invention is generally sufficient to ensure a satisfactory level of inhibition against dust formation / erosion. However, according to another embodiment, steps (b) to (d) or (b) and (c) may be repeated once, twice, three times or more when necessary to ensure particularly effective biobonding of the substrate to be treated, and thus particularly effective inhibition of dust formation / erosion.

[0184] According to another preferred embodiment of the method of the present invention, step (c) is performed once or repeatedly, and the total amount of component (iii) applied is at least 20g, preferably at least 40g, more preferably at least 60g, more preferably at least 80g, more preferably at least 100g, and most preferably at least 120g (based on an application area of ​​1 square meter in each case), and / or

[0185] The total amount of ingredient (iii) applied is up to 2000g, preferably up to 1600g, more preferably up to 1200g, more preferably up to 800g, more preferably up to 600g, and most preferably up to 400g (based on an application area of ​​1 square meter in each case).

[0186] As for the remainder, reference is made to observations regarding the use of the invention and the mixtures thereof, which are correspondingly effective for the method of the invention.

[0187] Another aspect of the invention relates to bio-cementable mixtures as defined herein (especially in the context of the use according to the invention or the method of the invention).

[0188] In the preferred mixture, component (iii) is selected from the group consisting of the following substances:

[0189] Albumin; starch ethers, alanine, lysine, styrene-acrylates, especially styrene-acrylate dispersions; ethylene-vinyl acetate, especially ethylene-vinyl acetate dispersions; polyvinyl alcohol; magnesium sulfate; polyvinyl acetate, especially polyvinyl acetate dispersions; styrene-butadiene, especially styrene-butadiene dispersions; humic acid and combinations thereof, and polymers containing monomers of the above polymers.

[0190] Furthermore, in the preferred mixture, component (ii) is selected from the group consisting of the following substances:

[0191] Urea and its salts; organic acids such as lactic acid and its salts, preferably carboxylates and their esters; gluconic acid and its salts, preferably carboxylates and their esters; acetic acid and its salts, preferably carboxylates and their esters; formic acid and its salts, preferably carboxylates and their esters; propionic acid and its salts, preferably carboxylates and their esters; butyric acid and its salts, preferably carboxylates and their esters; valeric acid and its salts, preferably carboxylates and their esters; peptides, preferably containing non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine and / or glutamic acid; amino acids, preferably non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine and glutamic acid and their salts, preferably carboxylates and their esters; plant and animal composite matrices, especially peptones, yeast extracts, meat extracts, nutrient broths and casein amino acids; industrial residual substrates, especially corn extract, lactose mother liquor, protein lysates, preferably derived from peas, meat, potatoes or tomatoes; anaerobic substances, preferably carbon dioxide and methane.

[0192] In this document, the salt used as component (ii) is preferably its respective calcium salt. The advantage of doing so is that component (ii) can serve both as a substance for forming carbonates and as a preferred cation source according to optional component (iv).

[0193] For the remainder, reference is made to observations regarding the use and method according to the invention, which are correspondingly valid for the mixtures according to the invention.

[0194] When the bio-cementing mixtures of the present invention were tested on various substrates, more unexpected observations were obtained, which will be described in more detail below. These observations led to other potential areas of use representing other aspects of the invention.

[0195] Therefore, another aspect of the invention relates to the use of mixtures suitable for bio-binding for granulation, the mixture comprising, or consisting of, the components (i), (ii) and (iii) as specified herein, and optional components (iv) and / or optional components (v).

[0196] When the mixture of the present invention was applied to mobile samples of iron ore, small agglomerates or particles were found to form after application. The particles formed in the granulator exhibited higher strength than the control mixture (and reduced emissions during production).

[0197] It turned out that the granules produced using the control mixture were difficult to process because they were very fragile. This is likely due to the lack of viscosity-modifying compounds. Therefore, viscosity-modifying materials can also be used to produce bio-binders that can be used for granulation.

[0198] Surprisingly, some matrices dried more slowly in the granulator after the bio-binding agent was formed; therefore, the effect of the bio-binding mixture on evaporation was investigated in greater depth.

[0199] Therefore, another aspect of the invention relates to the use of mixtures suitable for biocementation for controlling, and in particular reducing, evaporation, the mixture comprising, or consisting of, the components (i), (ii) and (iii) as specified herein, and optional components (iv) and / or optional components (v).

[0200] The inventors discovered that the effective forming layer reduces the drying rate of sand. This is evident from the higher relative soil moisture of samples using the mixture of the present invention compared to those using water or control mixtures not of the present invention. It is assumed that the resulting bio-cementing layer comprises a viscosity barrier against downward-flowing water. It can be assumed that, considering the presence of viscosity-modifying substances, the porosity of this layer is altered to allow water to evaporate at a slower rate.

[0201] The altered porosity can also be used in other applications where porosity plays a role. This is particularly applicable to insulating materials, catalyst beds, and / or battery materials. Accordingly, another aspect of the invention relates to the use of mixtures suitable for biocementation in the production of insulating materials, catalyst beds, and / or battery materials, the mixture comprising, or consisting of, components (i), (ii), and (iii) as specified herein, and optional components (iv) and / or optional components (v).

[0202] Due to the reduced porosity, materials based on the mixtures of the present invention are also particularly suitable as sealing materials. Therefore, another aspect of the invention relates to the use of mixtures suitable for bio-bonding in the production of sealing materials, the mixture comprising, or consisting of, components (i), (ii), and (iii) as specified herein, and optional components (iv) and / or optional components (v).

[0203] Another aspect of the invention relates to the use of mixtures suitable for biocementing for the (heavy metal ion) decontamination and / or (heavy metal ion) precipitation from a matrix, the mixture comprising, or consisting of, components (i), (ii) and (iii) as specified herein, and optional components (iv) and / or optional components (v).

[0204] Those skilled in the art will recognize that, for example, carbonate ions produced by bacteria from urea can be used to precipitate metal ions (Phillips et al., Engineering applications of urea decomposition and biomineralization: a review, Biodeposition, 2013, Vol. 29, No. 6, 715–733). It is presumed that, given this, layer formation begins more rapidly when using soil contaminated with heavy metals. Therefore, tests were conducted to determine whether viscosity modifiers were also suitable for improving the precipitation of heavy metal ions. In experiments, the inventors were able to demonstrate that the mixtures according to the invention can bind and precipitate heavy metal ions.

[0205] Another aspect of the invention relates to the use of mixtures suitable for biocementing for the (heavy metal ion) detergency and / or (heavy metal ion) precipitation of solutions, the mixture comprising, or consisting of, components (i), (ii) and (iii) as specified herein, and optional components (iv) and / or optional components (v).

[0206] The viscosity-modifying compound in the mixture according to the invention is characterized by its synergistic effect with microbial bio-binding to produce a high-viscosity, low-emission bio-binding agent. Unexpectedly, results show that this also occurs in solution, thus producing particularly efficient precipitation of heavy metal ions from solution. This is especially unexpected because polymers have a particular tendency to bind polyvalent ions, particularly including divalent metal cations such as Ca(II), Cu(II), Mg(II), and Ni(II), to disperse them in solution, thereby increasing their solubility. Due to this affinity for divalent metal cations, it is expected, especially if the water-soluble and / or water-dispersible viscosity-modifying compound is a polymer, that it will stabilize polyvalent metal cations and their aggregates and agglomerates in solution, leading to a reduction in the precipitation efficiency of metal ions (see Tadros TF 2016, Nanodispersions, ISBN-978-3-11-029033-2, especially Chap. p. 25ff stereostabilization).

[0207] Accelerating curing additives are also used in building materials, such as cementitious building materials like mortar and concrete, to regulate curing time (see, for example, EP 2664596 A3). Based on the accelerated curing of the mixtures according to the invention described in Example 1 below, another aspect of the invention is the use of bio-cementing mixtures suitable for the production of building materials, particularly cemented building materials like mortar and concrete, and / or for regulating their curing time, the mixture comprising, or consisting of, components (i), (ii), and (iii) as specified herein, and optional components (iv) and / or optional components (v).

[0208] The mixtures according to the invention have been further adapted to bind and aggregate wood chips. Accordingly, other aspects of the invention relate to the use of mixtures suitable for bio-bonding in the production of building materials, particularly adhesives for insulating materials, the mixtures comprising, or consisting of, components (i), (ii), and (iii) as specified herein, and optional components (iv) and / or optional components (v). The statements made herein with respect to uses according to the invention are equally valid for the methods of the invention described herein and the mixtures according to the invention described herein, and vice versa. This is particularly suitable for (preferred) embodiments of uses according to the invention, which form corresponding (preferred) embodiments of the methods of the invention, and corresponding (preferred) embodiments of mixtures according to the invention, and vice versa.

[0209] The invention will now be described in more detail through selected embodiments. Unless otherwise stated, all data relate to weight.

[0210] Figure 1 Mechanical and dust suppression properties of the bio-adhesive and reference mixtures used with *S. pasteurii*: Penetration depth (in mm) of metal cones of different weights after 24 hours of reaction (top left). Tensile strength (in Newtons) of the resulting layer after 48 hours of reaction (top right). Emission-related weight loss after 24 hours of reaction following 1 minute exposure in a 12 m / s wind (bottom left). Emission-related weight loss after 24 hours of reaction, and penetration depth determination of a 600 g metal cone after 1 minute of exposure in a 12 m / s wind (bottom right).

[0211] Figure 2 Mechanical and dust suppression properties of the bio-adhesive and reference mixtures using *L. sphaericus*: Penetration depth (mm) of metal cones of different weights after 24 hours of reaction (top left). Fracturing strength (Newtons) of the resulting layer after 48 hours of reaction (top right). Emission-related weight loss after 24 hours of reaction following 1 minute of exposure at 12 m / s wind (bottom left). Emission-related weight loss after 2 hours of reaction, and penetration depth of a 600 g metal cone after 1 minute of exposure at 12 m / s wind (bottom right).

[0212] Figure 3Mechanical and dust suppression properties of the bio-adhesive and reference mixtures used for various bacterial strains: Penetration depth (in mm) of metal cones of different weights after 24 hours of reaction (top left). Weight loss after 24 hours of reaction, and penetration depth of a 600g metal cone after 1 minute of exposure to a 12m / s wind (top right). Tensile strength (in Newtons) of the resulting layer after 48 hours of reaction (bottom left). Weight loss after 48 hours of reaction, and tensile strength after 1 minute of exposure to a 12m / s wind (bottom right).

[0213] Figure 4 Emission-related weight loss of reference mixtures R1, R2 and R8 and bio-cemented mixture M20 after 24 hours of reaction time and 15 minutes of exposure to wind (6 m / s).

[0214] Figure 5 Demonstration of dust suppression effects at a limestone quarry, with aerial views of three application areas: road (1), fresh waste dump (2), and mine (3) (top left). Implementation of water spraying by a sprinkler truck as a current dust suppression measure (top right). Application of the mixture to the road (bottom left) and waste dump (bottom right).

[0215] Figure 6 Mechanical and dust suppression properties of biocement and reference mixtures when using *S. pasteurii*: breaking strength after four-day reaction time (top). Emission-related weight loss, breaking strength, and determination after one minute of exposure in a 12 m / s wind after four-day reaction time (bottom). Reference mixture R3 is crucial for all mixtures in the figure. For clarity, the corresponding invented biocement mixture is always placed to the right of the relevant reference mixture. Neither R3 nor R7 reduces emissions after mechanical testing; the combination of the two has a very effective emission reduction characteristic (M24).

[0216] Figure 7 Mechanical and dust suppression properties of non-favorable biocementation and reference mixtures when using *S. pasteurii*: breaking strength after four-day reaction time (top). Emission-related weight loss, breaking strength, and determination after one minute of exposure in a 12 m / s wind after four-day reaction time (bottom).

[0217] Figure 8Other examples of the use of the inventive mixtures. Particles produced using mixtures M7, M8, and M9 (from left to right). The paper substrate frame size is 5 mm (top). R2 (hollow rhombus), R3 (cross), M11 (solid square), M16 (hollow triangle), and M22 (hollow circle) relative soil moisture of the treated samples over a 168-day observation period. The evaporation control of the inventive bio-binder is evident from the higher relative soil moisture. M11 and M22 are close to each other (middle). 24-hour reaction time and residual heavy metal ion content in the supernatant after subsequent centrifugation (bottom). Detailed Implementation

[0218] Example 1: Accelerated bio-binding (improved dust suppression)

[0219] Materials and methods:

[0220] The volume of the experiment in the laboratory is 450 cm³. 3 The application is carried out in a plastic container. The application area is 78.5 cm² in each case. 2 .

[0221] The soil matrix used in the experiment consisted of silica sand with a 0-2mm gradation. The sand had been washed and dried by the manufacturer and was used directly. 800g of silica sand was used as the soil matrix in each plastic container. The plastic containers were filled.

[0222] As a control, a reference mixture was used, which consisted of the following components at the following concentrations:

[0223] Reference Mixture 1 (R1): Dry sand matrix, without added aqueous components.

[0224] Reference mixture 2 (R2): Water applied.

[0225] Reference mixture 3 (R3):

[0226] 48g / L urea

[0227] 44g / L calcium chloride

[0228] 4 x 10^8 cells / mL *S. pasteurii*

[0229] Reference mixture 4 (R4):

[0230] 6.25 g / L calcium lignosulfonate

[0231] Reference mixture 5 (R5)

[0232] 3.15 g / L calcium lignosulfonate

[0233] The biocementing reference system R3, modified from Stabnikov, V. et al., Water, Air & Soil Pollution (2013) 224:1631, was used for dust suppression. This publication investigated dust suppression trends at wind velocities of 0.39 m / s and lower. The wind velocities studied in this embodiment were significantly higher. The total amount of biocementing mixture provided in this embodiment was greater than four times. An exact replication of the reference did not produce significant changes relative to R3.

[0234] Mixture R3 also includes trace elements and trace amounts of salts and sugars (less than 1 wt%). Urea in this culture medium is primarily used as a carbonate source.

[0235] Apply the reference mixture three times repeatedly to the test area. The application rate is consistently 4 liters per square meter per replicate. Use a pipette for application. After application, smooth the surface with a spatula. The reported measurements are the average of the three replicates, typically within 10% of the determined value.

[0236] A liquid bio-binding mixture is used, which consists of the following components at the following concentrations:

[0237] Mixture 1 (M1):

[0238] 48g / L urea

[0239] 44g / L calcium chloride

[0240] 6.25 g / L calcium lignosulfonate

[0241] 4 x 10^8 cells / mL *S. pasteurii*

[0242] Mixture 2 (M2):

[0243] 48g / L urea

[0244] 44g / L calcium chloride

[0245] 3.15 g / L calcium lignosulfonate

[0246] 4 x 10^8 cells / mL *S. pasteurii*

[0247] The mixture also includes trace elements and, for example, trace amounts of salts and sugars (less than 1 wt%). Urea in this culture medium is primarily used as a carbonate source. Calcium lignin sulfonate is a viscosity-modifying compound in mixtures M1 and M2.

[0248] Apply the mixture three times repeatedly to the test area. The application rate is consistently 4 liters per square meter per replicate. Use a pipette for application. After application, smooth the surface with a spatula. The reported measurements are the average of the three replicates, typically within 10% of the determined value.

[0249] Except for the strain *S. pasteurii*, all components of this mixture capable of biobinding are in solid form. As described, for example, in Cuthbert, MO et al., Ecological Engineering 2012, 41, 32-40 (see section 2.2 on page 33), the bacteria are present in liquid culture in a culture medium known in the art, using 5 g / L yeast extract in the context of this invention. The solid components and the bacteria in the liquid culture are mixed directly before use, and the solid components dissolve.

[0250] Following application of the reference mixture and the bio-gel mixture, incubation was conducted throughout the observation period (typically 28 days) under conditions of 20% to 60% atmospheric humidity and multiple ventilations per day. During this period, the lowest temperature observed was 14.2°C, and the highest temperature observed was 25.2°C.

[0251] After 24 hours, the penetration depth of the submerged cones of different weights (150g, 300g, and 600g) and their subsequent dust suppression effect in the wind tunnel were measured. According to the test standard method DIN EN 13279-2:2014-03 (Section 4.4.2.2), after 24 hours, a Vicat apparatus with a described submerged cone and release device (such as DIN EN 13279-2:2014-03) was used. Figure 2 and 3As described, the penetration depth of cones with different weights (total weight of the immersion cone and guide rod: 150g, 300g, 600g) was determined. For this purpose, the sample was placed under the immersion rod. The immersion rod was carefully lowered until it contacted the sample surface. It was held for two seconds, and the release device was activated. Under its own weight, the immersion cone penetrated the sample vertically. After the cone remained stationary for 5 seconds, the penetration depth was read on a scale. Sampling was performed at three test points at least 3 cm apart. The average value was obtained from the three determined values. The measured value fluctuated by no more than 10% around its absolute value. This measurement provides information about the hardening properties (see DIN EN 196-3, Section 6.3.1). After the measurement, the mass of the specimen (sample mass before exposure to wind) was determined, and the specimen was placed in a wind tunnel. The mechanically stressed specimen was exposed to a wind speed of 12 m / s for 1 minute. The airflow direction impacted the surface at an angle of 12.5°. After exposure to wind, determine the reduced mass (sample mass after wind exposure) and determine the emission-related weight loss according to the formula shown below. Subtract the weight of the sample container itself in each case.

[0252] The dust suppression effect was determined using individual samples in a wind tunnel without pre-existing mechanical stress: the mass of the hardened sample (sample mass before wind exposure) was measured, and the sample was placed in the wind tunnel. In the wind tunnel, airflow passed over the sample at a wind speed of 12 m / s for one minute. The airflow direction impacted the surface at an angle of 12.5°. The reduction in sample weight after wind exposure (sample mass after wind exposure) was measured, and the emission-related weight loss was determined using the following formula. The weight of the sample container itself was subtracted in each case.

[0253] Emission-related weight loss (in weight percentage) was measured as follows:

[0254] Emission-related weight loss = [(sample mass before exposure to wind)] 第xy天 -(Sample mass after exposure to wind) 第xy天 Sample mass before exposure to wind 第xy天 ]*100

[0255] After 48 hours, the fracture strength of the layer is determined. Fracture strength (maximum force measured) can be determined by the following method: This method is based on the standard test method for determining the strength of binders in DIN EN 196-1:2005-05. The fracture strength is measured using a digital (fracture) strength measuring instrument according to the manufacturer's instructions. The specimen is pressed into the sample (until the fracture point) using a crank-operated test bench, and the applied force is continuously measured. The average fracture strength is calculated through multiple measurements (>3). The average fracture strength is preferably from 0.5 N to 1000 N, more preferably from 1 N to 300 N.

[0256] After determining the fracture strength, the mechanically stressed sample was placed in a wind tunnel and exposed to a wind speed of 12 m / s for 1 minute. The airflow direction impacted the surface at a 12.5° angle. The emission-related weight loss was determined using the formula described above. This test can serve as a reference for the long-term stability of the sample and its dust suppression.

[0257] result:

[0258] When dry silica sand (R1) was exposed to a wind tunnel at a wind speed of 12 m / s for 1 minute, more than 50% of its weight was carried away as dust. In a moist state, under the same wind speed and exposure time, the sand further lost 1.12 wt% of its own weight as dust (R2). Under the given conditions, the sand was completely dried after 4.5 days. In this case, the dust suppression effect continuously decreased (data not shown). In the completely dried sample, the emission-related weight loss percentage was greater than 50% after 1 minute of wind exposure.

[0259] The invented formulations M1 and M2 exhibited faster curing properties than the reference systems R1, R2, R3, R4, and R5. After 24 hours, a 150g cone penetrated 9mm to 25mm into the reference system, while a 150g cone penetrated 2mm and 6mm into the invented formulations, respectively. Figure 1 (Top left). The same trend is also evident in heavier cones ( Figure 1 (top left).

[0260] After 24 hours, without prior mechanical stress, and after 1 minute of exposure in a wind tunnel (12 m / s), the emission-related weight loss was greater than 50% for R1, 1.11% for R2, 0.41% for R3, 0.66% for R4, and 0.99% for R5. The emission-related weight loss for mixture M1 was only 0.03%, and for mixture M2 it was 0.04%. Figure 1 (Lower left). This is likely due to better viscosity, which is also reflected in mechanical properties.

[0261] After 24 hours, under pre-stressed conditions (verifying the penetration depth of a 600g cone), and after 1 minute of exposure in a wind tunnel (12m / s), the emission-related weight loss was greater than 50% for R1, 1.12% for R2, 0.71% for R3, 1.58% for R4, and 2.78% for R5. The emission-related weight loss for mixture M1 was 0.16%, and the emission-related weight loss for mixture M2 was 0.18%. Figure 1 (bottom right).

[0262] After a 48-hour reaction time, the invented mixtures M1 and M2 exhibited higher fracture strengths than the relevant reference system. In this case, the fracture strength of the invented mixtures was higher than the sum of the individual components: fracture strength of R3 = 1.5 N, fracture strength of R5 = 2.2 N, fracture strength of M2 = 7 N. The fracture strength of R3 = 1.5 N, fracture strength of R4 = 5.1 N, fracture strength of M1 = 12 N. Figure 1 (top right).

[0263] As the samples aged, the differences in dust suppression effects after pre-mechanical verification became even more pronounced: after 48 hours, and 1 minute of exposure to wind in a wind tunnel (12 m / s) with pre-mechanical stress (measurement of fracture strength), the emission-related weight loss was 1.30% for R2, 0.85% for R3, 40.1% for R4, and 42.9% for R5. Mixtures M1 and M2 showed significantly lower emission-related weight losses, with M1 at 0.40% and M2 at 0.43%. The same trend was evident even after longer reaction times (10 days and 28 days, respectively) (data not shown; see also Example 2).

[0264] Bio-cemented mixtures advantageously exhibit effects similar to many standard commercial dust-suppressing compositions (data not shown), with the aforementioned increased dust suppression after mechanical loading comparable to that of bituminous systems, without the various environmental drawbacks.

[0265] Furthermore, in the aforementioned biocemented mixtures R3, M1, and M2, the bacterial strain *S. pasteurii* was replaced with the same cell count concentration of *L. sphaericus* in each case, and the experiments were performed in each case as described above. The resulting liquid reference and biocemented mixture consisted of the following components:

[0266] Reference mixture 6 (R6):

[0267] 48g / L urea

[0268] 44g / L calcium chloride

[0269] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0270] Mixture 3 (M3):

[0271] 48g / L urea

[0272] 44g / L calcium chloride

[0273] 6.25 g / L calcium lignosulfonate

[0274] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0275] Mixture 4 (M4):

[0276] 48g / L urea

[0277] 44g / L calcium chloride

[0278] 3.15 g / L calcium lignosulfonate

[0279] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0280] The mixture also contains trace elements and, for example, trace amounts of salts and sugars (less than 1 wt%). Urea in the culture medium is primarily used as a carbonate source. As described, for example, in Dick, J. et al., Biodegradation 2006, 17, 357-367 (see the “Materials and Methods” section, page 359), the bacteria are present in liquid cultures in media known in the art, using 5 g / L yeast extract in the context of this invention. Calcium lignin sulfonate is a viscosity-modifying compound in mixtures M3 and M4.

[0281] Using the organism *L. sphaericus* in the invented mixture yielded results comparable to those obtained using *S. pasteurii* (see [link to original text]). Figure 2 ).

[0282] Slightly modified formulations using bio-binding mixtures M1, M2, M3, and M4 also achieved considerable emission reduction effects. These formulations contained calcium acetate, calcium propionate, calcium formate, calcium lactate, and / or calcium chloride at concentrations ranging from 0.05M to 0.3M, with a total calcium concentration not exceeding 0.4M (data not shown). Larger variations in the concentrations of calcium lignosulfonate (e.g., 1 g / L to 500 g / L), urea (e.g., 0.1M to 1.0M), or yeast extract (e.g., 0.1 g / L to 30 g / L) also produced good emission reduction effects. In each case, dust suppression depended on the concentration of the respective bio-binding mixture component used (data not shown). Corresponding observations were also made on the corresponding modified formulations of Examples 2, 3, and 4.

[0283] Accelerators are used to regulate the curing time in building materials such as cementitious building materials, like mortar and concrete (EP2664596A3). Based on the accelerated curing properties of the mixtures described in this embodiment, the preferred use of the mixtures of the invention is in the production of building materials.

[0284] Similarly, considerable emission reduction effects were achieved for mixtures in which all the aforementioned bacteria existed in powder form. For this purpose, the corresponding bacterial cells were concentrated in a culture medium, and then professionally dried and dissolved before being applied to the respective culture medium.

[0285] Similarly, in mixtures in which calcium lignosulfonate is replaced by lignosulfonic acid, sodium lignosulfonate, potassium lignosulfonate, or ammonium lignosulfonate, and / or in mixtures in which a cation source is removed (in this article: calcium source, such as calcium chloride), all of the above mixtures achieve comparable emission reductions.

[0286] Example 2: Accelerated bio-binding formulation (powder) for use in non-urea decomposition and urea decomposition bio-binding systems Comparison of (dust suppression improved)

[0287] Materials and methods:

[0288] The volume of the experiment in the laboratory is 450 cm³. 3 The application is carried out in a plastic container. The application area is 78.5 cm² in each case. 2 .

[0289] The soil matrix used in the experiment consisted of silica sand with a 0-2mm gradation. The sand had been washed and dried by the manufacturer and was used directly. 800g of silica sand was used as the soil matrix for each plastic container.

[0290] As a control, reference mixture R3 was used, which consisted of the following components at the following concentrations:

[0291] Reference mixture 3 (R3):

[0292] 48g / L urea

[0293] 44g / L calcium chloride

[0294] 4 x 10^8 cells / mL *S. pasteurii*

[0295] Reference mixture 6 (R6):

[0296] 48g / L urea

[0297] 44g / L calcium chloride

[0298] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0299] Mixtures R3 and R6 also include trace elements and trace amounts of salts and sugars (less than 1 wt%). Urea in this culture medium is primarily used as a carbonate source.

[0300] Apply the reference mixture three times to the experimental area. The application rate is consistently 4 liters per square meter per repeat. Use a pipette for application. After application, smooth the surface with a spatula.

[0301] A liquid bio-binding mixture is used, which consists of the following components at the following concentrations:

[0302] Mixture 5 (M5):

[0303] 5g / L yeast extract

[0304] 21g / L calcium acetate

[0305] 34.9 g / L calcium chloride

[0306] 46.2 g / L calcium lactate

[0307] 0.40 g / L sodium hydroxide

[0308] 1.07 g / L ammonium chloride

[0309] 15g / L L-alanine

[0310] 25g / L calcium lignosulfonate

[0311] 4 x 10^8 cells / mL of *Bacillus pseudofirmus*

[0312] Mixture 6 (M6):

[0313] 27g / L urea

[0314] 34g / L calcium chloride

[0315] 10g / L yeast extract

[0316] 12.5 g / L styrene-acrylate dispersion

[0317] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0318] Mixture 7 (M7):

[0319] 5g / L yeast extract

[0320] 21g / L calcium acetate

[0321] 34.9 g / L calcium chloride

[0322] 46.2 g / L calcium lactate

[0323] 25g / L calcium lignosulfonate

[0324] 4 x 10^8 cells / mL halodurans

[0325] Mixture 8 (M8):

[0326] 36g / L urea

[0327] 35g / L calcium chloride

[0328] 10g / L yeast extract

[0329] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0330] Mixture 9 (M9):

[0331] 27g / L urea

[0332] 17g / L calcium chloride

[0333] 31g / L albumin

[0334] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0335] Mixture 10 (M10):

[0336] 48g / L urea

[0337] 44g / L calcium chloride

[0338] 3.9 g / L polyvinyl alcohol

[0339] 4 x 10^8 cells / mL *S. pasteurii*

[0340] Mixture 11 (M11):

[0341] 48g / L urea

[0342] 44g / L calcium chloride

[0343] 3.9 g / L polyvinyl acetate dispersion

[0344] 4 x 10^8 cells / mL *S. pasteurii*

[0345] Mixture 12 (M12):

[0346] 48g / L urea

[0347] 44g / L calcium chloride

[0348] 9.4 g / L starch ether

[0349] 4 x 10^8 cells / mL *S. pasteurii*

[0350] Mixture 13 (M13):

[0351] 1g / L yeast extract

[0352] 34.9 g / L calcium chloride

[0353] 25g / L calcium lignosulfonate

[0354] 21g / L calcium acetate

[0355] 46.2 g / L calcium lactate

[0356] 4 x 10^8 cells / mL Bacillus cohnii

[0357] Mixture 14 (M14):

[0358] 1g / L yeast extract

[0359] 21g / L calcium acetate

[0360] 25g / L calcium lignosulfonate

[0361] 15g / L L-alanine

[0362] 34.9 g / L calcium chloride

[0363] 46.2 g / L calcium lactate

[0364] 0.40 g / L sodium hydroxide

[0365] 1.07 g / L ammonium chloride

[0366] 4 x 10^8 cells / mL *A. crystallopoietes*

[0367] Mixture 15 (M15):

[0368] 1g / L yeast extract

[0369] 34.9 g / L calcium chloride

[0370] 21g / L calcium acetate

[0371] 46.2 g / L calcium lactate

[0372] 4 x 10^8 cells / mL Bacillus cohnii

[0373] Mixture 16 (M16):

[0374] 1.07 g / L ammonium chloride

[0375] 21g / L calcium acetate

[0376] 15g / L L-alanine

[0377] 34.9 g / L calcium chloride

[0378] 0.40 g / L sodium hydroxide

[0379] 1g / L yeast extract

[0380] 46.2 g / L calcium lactate

[0381] 4 x 10^8 cells / mL *A. crystallopoietes*

[0382] Mixture 17 (M17):

[0383] 36g / L urea

[0384] 36g / L magnesium sulfate

[0385] 10g / L yeast extract

[0386] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0387] Mixture 18 (M18):

[0388] 27g / L urea

[0389] 35g / L calcium chloride

[0390] 45g / L lysine

[0391] 4 x 10^8 cells / mL *S. pasteurii*

[0392] Mixture 19 (M19):

[0393] 48g / L urea

[0394] 44g / L calcium chloride

[0395] 25g / L polyvinyl alcohol

[0396] 4 x 10^8 cells / mL *S. pasteurii*

[0397] Mixture 20 (M20):

[0398] 27g / L urea

[0399] 47g / L calcium lignosulfonate

[0400] 4 x 10^8 cells / mL *S. pasteurii*

[0401] Mixture 21 (M21):

[0402] 5g / L yeast extract

[0403] 21g / L calcium acetate

[0404] 25g / L calcium lignosulfonate

[0405] 34.9 g / L calcium chloride

[0406] 46.2 g / L calcium lactate

[0407] 4 x 10^8 cells / mL of *Bacillus pseudofirmus*

[0408] Mixture 22 (M22):

[0409] 27g / L urea

[0410] 47g / L calcium lignosulfonate

[0411] 12g / L calcium chloride

[0412] 4 x 10^8 cells / mL *S. pasteurii*

[0413] Mixture 23 (M23):

[0414] 27g / L urea

[0415] 35g / L calcium chloride

[0416] 45g / L lysine

[0417] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0418] The mixtures also include trace elements and, for example, trace amounts of salts and sugars (less than 1 wt%). In mixtures M6, M8, M9, M10, M11, M12, M17, M18, M19, M20, M21, M22, and M23, urea is primarily used as a carbonate source. In mixtures M5, M7, M13, M14, M15, M16, and M21, organic calcium salts are primarily used as a carbonate source in each case. Additionally, in mixture M15, yeast extract is also used as a carbonate source. Furthermore, in mixture M16, L-alanine is used as another carbonate source.

[0419] L-alanine, calcium lignosulfonate, calcium lactate (M5, M7), calcium acetate (M1, M14), styrene-acrylate dispersion, yeast extract, albumin, polyvinyl alcohol, polyvinyl acetate dispersion, starch ether, magnesium sulfate, and lysine are viscosity-modifying compounds in this example (if they are not used as carbonate sources).

[0420] Apart from bacteria, all bio-binding components in this mixture are in solid form. For example, as described in Jonkers H.M. et al., Custom Concrete Structures – Walraven & Stoelhorst (ed.), 2008, Taylor & Francis Group, London, ISBN 978-0-415-47535-8, Section 2.1, the strain *Bacillus pseudofirmus* exists in liquid culture in media known in the art; in the context of this invention, 5 g / L yeast extract is used. Bacillus cohnii and halodurans were present in the same culture medium as Bacillus pseudofirmus, while A. crystallopoietes were present in known culture media, such as those described, for example, in Hamilton, RW et al., Journal of Bacteriology 1977, 129(2), 874-879 (see “Materials and Methods” section, pp. 874-875). Sphaericus spheroides and Pasteurii were present in the culture medium described in Example 1. The bacteria in the solid and liquid cultures were mixed directly before use, and the solid components were dissolved.

[0421] Apply the mixture three times repeatedly to the test area. The application rate is consistently 4 liters per square meter per replicate. Use a pipette for application. After application, smooth the surface with a spatula. The reported measurements are the average of the three replicates, typically within 10% of the determined value.

[0422] Following application of the reference mixture and the bio-gel mixture, incubation was conducted throughout the observation period (28 days) under conditions of 20% to 60% atmospheric humidity and multiple ventilations per day. During this period, the lowest temperature observed was 14.2°C, and the highest temperature observed was 25.2°C.

[0423] After 24 hours, as described in Example 1, the penetration depth of immersion cones of different weights (150g, 300g, and 600g) and their subsequent dust suppression effect in a wind tunnel were measured. Dust suppression was assessed using individual samples in a wind tunnel without pre-stressed mechanical stress: the cured samples were exposed to a wind speed of 12 m / s for 1 minute. The airflow impinged on the surface at a 12.5° angle. Emission-related weight loss (in weight percentage) was determined as described in Example 1.

[0424] After 48 hours, the fracture strength of the layer is determined. Fracture strength (maximum force measured) can be determined by the following method: This method is based on the standard test method for determining the strength of binders in DIN EN 196-1:2005-05. The fracture strength is measured using a digital (fracture) strength measuring instrument according to the manufacturer's instructions. The specimen is pressed into the sample (until the fracture point) using a crank-operated test bench, and the applied force is continuously measured. The average fracture strength is calculated through multiple measurements (>3). The average fracture strength is preferably from 0.5 N to 1000 N, more preferably from 1 N to 300 N.

[0425] After determining the fracture strength, the mechanically stressed sample was placed in a wind tunnel and exposed to a wind speed of 12 m / s for 1 minute. The airflow direction impacted the surface at a 12.5° angle. Emission-related weight loss was determined using the formula described in Example 1. This test can serve as a reference for the long-term stability of the sample and its dust suppression.

[0426] As described above, on the selected samples, the fracture strength and mass loss upon wind exposure were measured after 10 days and 28 days of reaction time, respectively.

[0427] result:

[0428] In Example 1 above, the reduction of emission-related weight loss was described using an accelerated biocementing formulation. This example illustrates how this finding can be extended to a wide range of soil consolidation materials that accelerate biocementation.

[0429] All the formulations of the above inventions exhibited faster curing properties than reference systems R3 and R6. After 24 hours, a 150g cone penetrated reference systems R3 and R6 to 14mm, while a 150g cone penetrated the formulations of the inventions to 9.5mm. Figure 3 (Top left). The same trend is also evident in heavier cones (top left). Figure 3 (top left).

[0430] If weight loss in a wind tunnel is measured after mechanical verification, the mixture of the present invention exhibits enhanced viscosity, thereby improving dust suppression. Weight loss after 24 hours of reaction time, penetration depth measurement, and exposure in a wind tunnel (12 m / s) for one minute are also considered. Figure 3 As shown in the upper right corner. In the case of reference system R3, the emission-related weight loss percentage is 0.71%. The mass loss of the inventive formulation is 0.07% to 0.56%. Figure 3 (Top right). Due to its faster hardening properties, exposure to wind results in fewer particles being carried away from the sample.

[0431] After a 48-hour reaction time, the inventive mixture exhibited a higher fracture strength than the relevant reference system. In this case, the fracture strength of the inventive mixture was a multiple of the fracture strength of the reference system R3. Figure 3 , lower left).

[0432] As the samples aged, the differences in dust suppression effects after previous mechanical verification became even more pronounced: after 48 hours, the weight loss of R2 was 1.30% and that of R3 was 0.85% after 1 minute of exposure to wind in a wind tunnel (12 m / s) with pre-stressed mechanical stress (for determining fracture strength).

[0433] The mixtures M15 to M23 of the invention showed a mass loss of 0.04% to 0.45% after mechanical testing and wind exposure (see [link]). Figure 3 (bottom right).

[0434] If the fracture mechanical properties and emission-related weight loss in the wind tunnel are studied after a long reaction period, the differences between the reference system and the bio-cemented mixture become even more apparent:

[0435] In addition, reference mixture R7 and bio-cemented mixture M24 were prepared and compared with each other as described above.

[0436] Reference mixture 7 (R7):

[0437] 50g / L calcium lignosulfonate

[0438] Mixture 24 (M24):

[0439] 48g / L urea

[0440] 44g / L calcium chloride

[0441] 50g / L calcium lignosulfonate

[0442] 4 x 10^8 cells / mL *S. pasteurii*

[0443] In mixture M24, trace elements and trace amounts of salts and sugars (less than 1 wt%) are also present. Calcium lignin sulfonate is a viscosity-modifying compound in mixture M24. The strain *S. pasteurii* was present in the culture medium described in Example 1. The mixture was prepared and stored as previously described.

[0444] These mixtures were found to solidify within 48 hours (fracture strength not shown; see also Example 5). For R7, a thin layer formed, while for M24, a thicker, more viscous layer formed. These differences in layer properties are reflected in the differences in emission-related weight loss. After determining the fracture strength, mixtures R7 and M24 were tested in a wind tunnel for 1 minute at a wind speed of 12 m / s (as described above). The emission-related weight loss for R7 was 11.3%, and for M24 it was 0.21%. The selected mixtures and the reference system were allowed to react for 48 hours. The results demonstrate to those skilled in the art that the enhanced viscosity offers advantages in persistent dust suppression.

[0445] The fracture strength of various reagents was measured after 10 and 28 days, compared with the reference system. The results obtained in this case were comparable to those described above (data not shown). The mass loss after mechanical verification after 10 days is shown in Table 1. This study found that the more viscous bio-binder layer exhibited significantly better dust suppression after mechanical testing.

[0446] Table 1: Mass loss of various reference mixtures and bio-cemented mixtures after 10-day reaction time, mechanical testing, and 1 minute of wind exposure at 12 m / s wind speed.

[0447]

[0448]

[0449] Comparable emission reductions were also achieved using slightly modified formulations of bio-cementing mixtures M5 to M24, which contain calcium acetate, calcium propionate, calcium formate, calcium lactate, and / or calcium chloride at concentrations of 0.05 M to 0.4 M in each case, with a total calcium concentration not exceeding 1 M (data not shown). Larger variations in the concentrations of calcium lignosulfonate (e.g., 1 g / L to 500 g / L), L-alanine (e.g., 1 g / L to 250 g / L), styrene-acrylate dispersion (e.g., 1 g / L to 350 g / L), polyvinyl alcohol (e.g., 1 g / L to 250 g / L), polyvinyl acetate dispersion (e.g., 1 g / L to 350 g / L), albumin (1 g / L to 200 g / L), starch ether (e.g., 1 g / L to 90 g / L), magnesium sulfate (e.g., 1 g / L to 300 g / L), lysine (e.g., 1 g / L to 250 g / L), urea (e.g., 0.1 M to 1.0 M), or yeast extract (e.g., 0.1 g / L to 150 g / L) also produce good emission reduction effects. In each case, dust suppression depends on the concentration of the respective components of the bio-cementing mixture used (data not shown). The corresponding modified formulations of Examples 3, 4 and 5 were also observed.

[0450] Comparable emission reductions were also achieved using bio-binding mixtures M5 to M24, in which the bacteria were present in the form of spray-dried and / or freeze-dried powder. For this purpose, the corresponding bacterial cells were concentrated in a culture medium, then professionally dried and dissolved in the appropriate culture medium prior to application. Studies have found that when using dried bacterial cells, a further slight reduction in emission-related weight loss can actually be achieved (data not shown).

[0451] Similarly, in mixtures where calcium lignosulfonate was replaced with lignosulfonic acid, sodium lignosulfonate, potassium lignosulfonate, and ammonium lignosulfonate, respectively, comparable effects were achieved to all the aforementioned mixtures. Furthermore, comparable dust suppression effects were achieved when removing the cationic source (calcium source in this case) from mixtures M5, M6, M7, M8, M9, M10, M11, M12, M13, M14, M18, M19, M21, M22, and M23. Simultaneously, comparable effects were again obtained when calcium lignosulfonate was replaced with lignosulfonic acid, sodium lignosulfonate, potassium lignosulfonate, and ammonium lignosulfonate, respectively, and the cationic source (calcium source in this text) was removed.

[0452] Example 3: Analysis of selected mixtures and reference systems in a wind tunnel of an external testing laboratory

[0453] Materials and methods:

[0454] In an external testing laboratory, the emission reduction effects of reference systems R1 (dry) and R2 (water applied) as well as commercially available dust inhibitor R8 were compared with those of mixture M20.

[0455] The soil matrix used was fine calcium carbonate, designated ESKAL 60. This fine particulate dust is used as test dust for various analyses, such as wind tunnel analysis. ESKAL 60 has a precisely defined particle size distribution. The average grain size is 60 μm. Those skilled in the art will recognize that the test dust used must be suitable for the wind tunnel in use. Plastic trays (87 mm in diameter, 16 mm in height) were filled to the rim with the soil matrix, and the precise weight of each container was determined.

[0456] Then, the appropriate surface treatment agent is provided to all samples. The treated samples are then marked in a blind test to prevent them from being assigned the appropriate surface treatment agent.

[0457] Reference Mixture 8 (R8): 50 g / L polymer dispersion (various types)

[0458] Commercially available products are shell-forming agents. Use according to manufacturer specifications, at a rate of 1.5 L / m³. 2 Application. Furthermore, the emission reduction effect of the bio-cemented mixture M20 was investigated. The mixture 20 was applied at a rate comparable to R8, and measurements were taken as the mass of solids per unit surface area.

[0459] Except for R2, all samples were equilibrated in a conditioning cabinet for 24 hours under defined environmental conditions (31% relative humidity, 23°C) and then weighed again. Reference mixture R2 was not applied to samples until exposure in the wind tunnel. Wetting application (R2) was performed immediately before the start of the experiment using deionized water in a uniformly positioned spray bottle. The water mass input was recorded.

[0460] At the start of the experiment, samples were randomly positioned and covered in the center of the wind tunnel (D = 0.15 m, L = 5.4 m). The particle counter was activated at the start of the experiment, the covering on the sample material was removed, and the wind tunnel was sealed. All samples were exposed to the airflow above them for 15 minutes (average aerosol velocity 6 m / s), and measurements were taken at the sample height (particle size distribution was determined every 30 seconds). All experiments were repeated three times. Emission-related mass loss was determined using the formula specified in Example 1. The reported measurements are the average of the three replicates, and the average is typically within 10% of the determined value.

[0461] result:

[0462] Experiments showed that surface treatment agent M20 reliably prevented dust from being carried away. Emissions occurred only in commercially available reagent R8, as well as in the untreated sample (R1) and the water-treated sample (R2).

[0463] With commercially available reagent R8, this behavior manifests as up to 180 captured particles within the first 90 seconds, with an average mass loss of 1.86%.

[0464] The untreated calcium carbonate sample (R1) used for comparison had the largest number of particles that could be carried away among all the samples examined. Emissions started at 2,500 to 3,800 particles per 30 seconds, increased to 4,100 to 5,500 particles per 30 seconds, and then steadily decreased to approximately 100 particles per 30 seconds. The emission-related average mass loss was 74.55%.

[0465] In the case of the water-treated samples, particle release was delayed; in this study, particle release began only after approximately 200 seconds. The emission-related average mass loss was 66.94%. The delayed release is likely due to moisture evaporation in the wind tunnel.

[0466] In the case of bio-cemented mixture M20, there was no detectable particle release and the emission-related mass loss was 0.003% (see [reference]). Figure 4 ).

[0467] Example 4: Open-air demonstration of emission reduction effect in limestone mines

[0468] Materials and methods:

[0469] To control dust suppression under open-pit conditions, bio-cementing mixture M20 was applied at three sites in a limestone quarry, compared to a reference mixture R3 (as a control). The three sites within the mine were located on roads ( Figure 5 Location 1 in the middle, upper left), fresh waste dump ( Figure 5 Location 2 in the middle, upper right) and in the active mine ( Figure 5 Site 3 in the middle, upper left). In each case, apply at 150m 2 The application was carried out over an area of ​​3 liters per square meter. As a further reference, emission reduction measures currently used in routine mine operations were implemented: 3 liters of water per square meter (…). Figure 5 (Top right). This was performed in the same manner as reference mixture R2. The reference area was directly adjacent to the test area of ​​the bio-cemented mixture, and the same operating level was observed. Mixture M20 was supplied to the road as shown in the example. Figure 5 As shown in the lower left corner; Figure 5 The lower right corner depicts the supply area for mixture M20 on the waste disposal site.

[0470] After application, such as Figure 5As shown at the bottom, all nine areas where the application was performed were pinned, allowing for a response within 48 hours. After 24 hours, the layer formation was visually assessed, and the fracture strength of the layer was measured after 48 hours (data not shown).

[0471] The outdoor experiment lasted for four weeks. During this period, the temperature varied between 5.3°C at night and 26.3°C during the day. The relative humidity varied between 64% at night and 31% during the day. The total precipitation during the experiment was 11 L / m³. 2 .

[0472] Dust suppression effectiveness was measured at different time points, including 48 hours, 7 days, and 28 days. After 48 hours, dust suppression effectiveness was verified at multiple points using a Bosch blade blower (GBL 18V-120). The wind speed used in this study was 40 m / s, 1 meter above the surface, and an incident angle of approximately 15 degrees. Inspections conducted by three mine employees were categorized as "heavy dust generation," "moderate dust generation," and "no dust generation." All employees were technicians specializing in area dust suppression in mining, each with over 10 years of relevant professional experience. "No dust generation" was used when no visible particles were removed. "Heavy dust generation" was used when the test area formed dust in the same manner as the untreated area. "Moderate dust generation" was used when dust formation was reduced compared to the untreated area. The professionally obtained data were also validated by particle analysis (data not shown).

[0473] After the initial test (48 hours), the study area was reopened for operation, and obstacles were removed. Caution was exercised at this point to ensure all areas received equal exposure. Visual inspections were conducted on all areas at 7 days, 28 days, and 48 hours, and dust suppression effectiveness was measured.

[0474] result

[0475] One day later, the accelerated bio-binding agent M20 layer was perceptible, while the layer of reference mixture 3 did not solidify. After a reaction time of 48 hours, the relative tensile strength of the layers as described in Example 2 could be reproduced (data not shown).

[0476] After 48 hours of dust suppression testing using Bosch (GBL 18V-120), experts gave the following rating:

[0477] Mixture 20 (M20) – “Dust-free”

[0478] Reference 3 (R3) – “Moderate dust generation”

[0479] Reference 2(R2) – “Moderate dust generation”.

[0480] In this case, there was no difference in dust suppression effect among the application sites.

[0481] Seven days after application, the three application sites—roads, waste dumps, and mine pits—were inspected according to the above plan. At the waste dumps and in the mine pits, it was evident that a significant solid layer remained in the case of mixture 20 (M20), while no layer was formed in the cases of reference mixtures R2 and R3. The dust suppression trend rating is as follows:

[0482] Mixture 20 (M20) – “Dust-free”

[0483] Reference 3 (R3) – “Moderate dust generation”

[0484] Reference 2 (R2) – “Severe dust generation”.

[0485] On roads, the effect is even more pronounced. This is due to the effectiveness of the invention, in which the invented bio-cementing mixture M20 has high mechanical strength. For roads, the following ratings were given:

[0486] Mixture 20 (M20) – “Dust-free”

[0487] Reference 3 (R3) – “Severe dust generation”.

[0488] Reference 2 (R2) – Severe dust generation.

[0489] After 28 days, the results were comparable to those obtained after 7 days. The experiment was then discontinued.

[0490] Similarly, in mixtures where bacteria are present in powder form, emission reduction effects comparable to those of all the aforementioned mixtures were achieved. For this purpose, the corresponding bacterial cells are concentrated in a culture medium, and then professionally dried and dissolved before being applied to the respective culture medium.

[0491] This embodiment impressively demonstrates that the invented formulation exhibits improved dust suppression under mechanical load due to its faster solidification and higher strength. Furthermore, the hardened shell produced using the invented formulation can be maintained for a longer period compared to existing systems.

[0492] Example 5: Synergistic effect of bio-binding agents and viscosity-modifying compounds

[0493] Materials and methods:

[0494] The volume of the experiment in the laboratory is 450 cm³. 3 The application is carried out in a plastic container. The application area is 78.5 cm² in each case. 2 .

[0495] The soil matrix used in the experiment consisted of silica sand with a 0-2mm gradation. The sand had been washed and dried by the manufacturer and was used directly. 800g of silica sand was used as the soil matrix in each plastic container. The plastic containers were filled.

[0496] As a control, a reference mixture was used, which consisted of the following components at the following concentrations:

[0497] Reference mixture 3 (R3):

[0498] 48g / L urea

[0499] 44g / L calcium chloride

[0500] 4 x 10^8 cells / mL *S. pasteurii*

[0501] Reference mixture 7 (R7):

[0502] 50g / L calcium lignosulfonate

[0503] Reference mixture 9 (R9)

[0504] 25g / L polyvinyl alcohol

[0505] Reference Mixture 10 (R10)

[0506] 15.6 g / L polyvinyl alcohol

[0507] Reference mixture 11 (R11):

[0508] 9.4 g / L starch ether

[0509] Reference mixture 12 (R12):

[0510] 50g / L humic acid

[0511] Reference mixture 13 (R13):

[0512] 50g / L sodium silicate

[0513] Reference mixture 14 (R14):

[0514] 25 g / L styrene-butadiene dispersion

[0515] Mixture R3 also includes trace elements and trace amounts of salts and sugars (less than 1 wt%). Urea in this culture medium is primarily used as a carbonate source.

[0516] Except for the styrene-butadiene dispersion, humic acid, and the strain *S. pasteurii*, all components of the bio-gelatinizable mixture of the invention are in solid form. As described, for example, in Cuthbert, MO et al., Ecological Engineering 2012, 41, 32-40 (see section 2.2 on page 33), the bacteria are present in a liquid culture in a culture medium known in the art, using 5 g / L yeast extract in the context of this invention. The solid components and the bacteria in the liquid culture are mixed directly before use, and the solid components dissolve.

[0517] Apply the reference mixture three times repeatedly to the experimental area. The application rate is always 4 liters per square meter per replicate. Apply the completely dissolved sample using a pipette. After application, smooth the surface with a spatula. The reported measurement is the average of the three replicates, typically within 10% of the determined value.

[0518] A liquid bio-binding mixture is used, which consists of the following components at the following concentrations:

[0519] Mixture 12 (M12):

[0520] 48g / L urea

[0521] 44g / L calcium chloride

[0522] 9.4 g / L starch ether

[0523] 4 x 10^8 cells / mL *S. pasteurii*

[0524] Mixture 19 (M19):

[0525] 48g / L urea

[0526] 44g / L calcium chloride

[0527] 25g / L polyvinyl alcohol

[0528] 4 x 10^8 cells / mL *S. pasteurii*

[0529] Mixture 24 (M24):

[0530] 48g / L urea

[0531] 44g / L calcium chloride

[0532] 50g / L calcium lignosulfonate

[0533] 4 x 10^8 cells / mL *S. pasteurii*

[0534] Mixture 25 (M25):

[0535] 48g / L urea

[0536] 44g / L calcium chloride

[0537] 15.6 g / L polyvinyl alcohol

[0538] 4 x 10^8 cells / mL *S. pasteurii*

[0539] Mixture 26 (M26):

[0540] 48g / L urea

[0541] 44g / L calcium chloride

[0542] 50g / L humic acid

[0543] 4 x 10^8 cells / mL *S. pasteurii*

[0544] Mixture 27 (M27):

[0545] 48g / L urea

[0546] 44g / L calcium chloride

[0547] 50g / L sodium silicate

[0548] 4 x 10^8 cells / mL *S. pasteurii*

[0549] Mixture 28 (M28):

[0550] 48g / L urea

[0551] 44g / L calcium chloride

[0552] 25 g / L styrene-butadiene dispersion

[0553] 4 x 10^8 cells / mL *S. pasteurii*

[0554] Mixtures M12, M19, M24, M25, M26, M27, and M28 also include trace elements and trace amounts of salts and sugars (less than 1 wt%). Urea in this culture medium is primarily used as a carbonate source.

[0555] Starch ether, polyvinyl alcohol, calcium lignosulfonate, humic acid (in each case a polymer), sodium silicate, and styrene-butadiene dispersions are viscosity-modifying compounds in mixtures M12, M19, M24, M25, M26, M27, and M28. Urea serves as a carbonate source in mixtures M12, M19, M24, M25, M26, M27, and M28.

[0556] Except for the styrene-butadiene dispersion, humic acid, and the strain *S. pasteurii*, all components of this bio-cementing mixture are in solid form. As described, for example, in Cuthbert, MO et al., Ecological Engineering 2012, 41, 32-40 (see section 2.2 on page 33), the bacteria are present in a liquid culture in a culture medium known in the art, using 5 g / L yeast extract in the context of this invention. The solid components and the bacteria in the liquid culture are mixed directly before use, and the solid components dissolve.

[0557] Apply the mixture three times repeatedly to the test area. The application rate is always 4 liters per square meter per repeat. Apply the completely dissolved sample using a pipette. After application, smooth the surface with a spatula. The reported measurement is the average of the three replicates, typically within 10% of the determined value.

[0558] Following application of the reference mixture and the bio-binding mixture, incubation was conducted for 28 days during the total observation period, under atmospheric humidity ranging from 20% to 60% and with multiple air changes per day. The lowest temperature observed during this period was 14.2°C, and the highest was 25.2°C.

[0559] After 1, 2, 3, 4, 10, and 28 days, fracture strength and emission-related weight loss were measured as described in Examples 1 and 2. Additionally, layer thickness was measured.

[0560] The fracture strength of each layer was determined after 1, 2, 3, 4, 10, and 28 days. The fracture strength (maximum force measured) was determined by the following method: This method is based on the standard test method for determining the strength of binders in DIN EN 196-1:2005-05. The fracture strength was measured using a digital (fracture) strength measuring instrument according to the manufacturer's instructions. The specimen was pressed into the sample (up to the fracture point) using a crank-operated test bench, and the applied force was continuously measured. The average fracture strength was calculated through multiple measurements (>3). The average fracture strength was preferably from 0.5 N to 1000 N, more preferably from 1 N to 300 N.

[0561] After determining the fracture strength, the thickness of the formed layer was measured. This was done manually using calipers after the layer mechanically fractured. The layer thickness was measured at six points on the fractured layer; the deviation of each measurement was 1 mm. The layer thickness was recorded as the arithmetic mean of the individual measurements.

[0562] After determining the layer thickness, the mechanically stressed sample was placed in a wind tunnel and exposed to a wind speed of 12 m / s for 1 minute. The airflow direction impacted the surface at a 12.5° angle. Emission-related weight loss was determined using the formula described in Example 1. This test can serve as a reference for the long-term stability of the sample and its dust suppression.

[0563] result:

[0564] In the preceding Examples 1 to 4, formulations exhibiting faster hardening properties and reduced emissions-related weight loss were described. During the analysis, it was unexpectedly found that for the mixtures of the invention, there is not necessarily a correlation between breaking strength and emissions reduction. Referencing reference mixtures R9 and R10 ( Figure 6 For the reagents described in the prior art, the correlation is indeed readily predictable and observable. After a four-day reaction time, the reference system yielded a breaking strength of R9 = 53.8 N and R10 = 29.8 N. Following mechanical verification, the emission-related weight losses for R9 and R10 were 3.79% and 7.72%, respectively. The more robust reference system was found to exhibit a lower emission-related weight loss. Therefore, a negative correlation exists between breaking strength and emission-related weight loss. A relative reduction in emission-related weight loss was observed in the calcium lignosulfonate reference system as breaking strength increased: where calcium lignosulfonate was used at 25 g / m² sand. 2 Up to 400g / m 2 The amount supplied increases the breaking strength linearly and reduces emission-related weight loss (data not shown).

[0565] In the case of the inventive mixture, after two days, no observable direct relationship was found between high breaking strength and low emission-related weight loss (see Example 2). For example, M24 had a breaking strength of 14 N after four days, while the relevant reference system R7 had a breaking strength of 26.5 N. However, R7 showed a significantly higher emission-related weight loss of 53%. At this point, reference system 3 showed almost no consolidation (breaking strength R3 = 1.5 N) and exhibited a 51% emission-related weight loss after breaking strength determination and exposure to wind. The combination of the two systems (mixture 24) produced an emission-reducing system with an emission-related weight loss of only 0.87%. The breaking strength of this system was M24 = 14 N. In the context of the prior art, it is not expected that such a mixture with lower breaking strength would have a significantly better dust suppression effect. This is attributed to the synergistic effect between biocementing and viscosity modifiers: M24, M19, M25, and M12 exhibited significantly lower emission-related weight losses than their individual components R3 and R7, R3 and R9, R3 and R10, and R3 and R11 ( Figure 6 The tensile strength of these mixtures is as follows: Figure 6 As shown above, and the emission-related weight loss after mechanical verification is as follows: Figure 6 As shown below. However, high tensile strength has no adverse effect on dust suppression and can be considered an additional advantage of bio-cementing mixtures in some cases (see Example 2). In this paper, the role of the viscosity modifier is in the fracture mode of the bio-cementing layer. After the layer fractures, R7 breaks into many small fragments, while in the case of M24, only small pores remain. These small fragments are easily blown away and dispersed by the wind.

[0566] The layer thickness measurements yielded the following values: the layer thickness of R7 was 8 mm, while the layer thickness of M24 was 14 mm.

[0567] Similar observations were made when using viscosity-modifying compounds such as starch ethers (R11, M12), humic acid (R12, M26), sodium silicate (R13, M27), and styrene-butadiene dispersions (R14, M28). In these mixtures, the corresponding bio-cemented mixtures exhibited lower breaking strengths than the corresponding reference breaking strengths, but with less emission (see [reference]). Figure 6 ).

[0568] The emission reduction effects comparable to those of mixtures M12, M19, M24, M25, M26, M27, and M28 also exist in the case of slightly modified formulations of bio-cemented mixtures containing calcium acetate, calcium propionate, calcium formate, calcium pyruvate, calcium salicylate, calcium citrate, and / or calcium chloride, in each case at concentrations from 0.05 M to 0.4 M, and not exceeding a total calcium concentration of 1 M (data not shown). Larger variations in the concentrations of calcium lignosulfonate (e.g., 1 g / L to 500 g / L), polyvinyl alcohol (e.g., 1 g / L to 250 g / L), starch ether (e.g., 1 g / L to 90 g / L), humic acid (e.g., 1 g / L to 350 g / L), potassium silicate and sodium silicate (e.g., 1 g / L to 450 g / L), polyvinyl alcohol, urea (e.g., 0.1 M to 1.0 M), or yeast extract (e.g., 0.1 g / L to 30 g / L) also resulted in effective emission reductions. In each case, dust suppression depended on the concentration of the respective biocementing mixture components used (data not shown). Comparable effects were also obtained when the bacterial strains were replaced with *L. sphaericus*, *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* at the same cell count per milliliter (data not shown). When *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* were used in the formulation in a similar manner at the same cell count per milliliter, the base components were further adapted to the requirements of the specific bacterial strains. In this document, those skilled in the art will recognize that, for these non-urea-degrading biocementing bacterial strains, the basal medium must be adjusted in a manner similar to that listed in Example 2, particularly in terms of suitable metabolic starting materials. The effect of the mixture on emission reduction was comparable to the results illustrated for *S. pasteurii* (data not shown).

[0569] Similarly, in mixtures where bacteria exist in powder form, the emission reduction effects of all the above mixtures are comparable. For this purpose, the corresponding bacterial cells are concentrated in a culture medium, then professionally dried and dissolved in the appropriate culture medium before application.

[0570] Similarly, in mixtures where calcium lignosulfonate was replaced with lignosulfonic acid, sodium lignosulfonate, potassium lignosulfonate, and ammonium lignosulfonate, respectively, comparable emission reduction effects as in mixture M24 were achieved. Removal of the cation source (in this context: calcium source, e.g., calcium chloride) from mixtures M12, M19, M24, M25, M26, M27, and M28 also yielded comparable dust suppression results. This also resulted in comparable dust suppression outcomes when both the replacement of lignin derivatives (as described above, e.g., with lignosulfonic acid) and the removal of the cation source (in this context: calcium source) were performed.

[0571] Based on these results, it is reasonable to assume that a cationic source, especially a calcium source, is optional when using the viscosity-modifying compounds disclosed herein.

[0572] Example 6: Applicable to reducing emission-related bio-binder weight loss and extending bio-binder integrity. Determination of minimum requirements for viscosity-modifying compounds

[0573] Materials and methods:

[0574] The volume of the experiment in the laboratory is 450 cm³. 3 The application is carried out in a plastic container. The application area is 78.5 cm² in each case. 2 .

[0575] The soil matrix used in the experiment consisted of silica sand with a 0-2mm gradation. The sand had been washed and dried by the manufacturer and was used directly. 800g of silica sand was used as the soil matrix in each plastic container. The plastic containers were filled.

[0576] As a control, a reference mixture was used, which consisted of the following components at the following concentrations:

[0577] Reference mixture 3 (R3):

[0578] 48g / L urea

[0579] 44g / L calcium chloride

[0580] 4 x 10^8 cells / mL *S. pasteurii*

[0581] Reference Mixture 15 (R15):

[0582] 50g / L polyvinyl acetate 20 (solid, granules)

[0583] Reference mixture 16 (R16):

[0584] 50g / L polycarbonate (solid, granules)

[0585] Reference mixture 17 (R17):

[0586] 50g / L vegetable oil (rapeseed oil)

[0587] Reference mixture 18 (R18):

[0588] 12.5g / L long-chain fatty acids (stearic acid)

[0589] Reference mixture 19 (R19):

[0590] 50 g / L starch, untreated (solid, powder)

[0591] The reference mixture contains compounds that are not water-soluble, water-dispersible, or water-emulsifiable, and does not contain component (iii). Mixture R3 also includes trace elements and, for example, trace amounts of salts and sugars (less than 1 wt%). Urea in this culture medium is primarily used as a carbonate source.

[0592] Except for the strain *S. pasteurii*, all components of this bio-gelatinizable mixture are in solid form. As described, for example, in Cuthbert, MO et al., Ecological Engineering 2012, 41, 32-40 (see Section 2.2 on page 33), the bacteria are present in liquid culture in media known in the art, using 5 g / L yeast extract in the context of this invention. The solid components and the bacteria in the liquid culture are mixed directly before use, dissolving the water-soluble solid components. The water-insoluble, non-dispersible, and non-emulsifiable substances are pre-coated uniformly onto the top layer of sand to achieve uniform application and eliminate any adverse effects of possible non-uniform application on dust suppression tests.

[0593] The reference mixture was applied three times repeatedly to the experimental area. The application rate was consistently 4 liters per square meter per replicate. The completely dissolved sample was applied using a pipette. After application, the surface was smoothed with a spatula. The reported measurements are the average of the three replicates, typically within 10% of the determined value.

[0594] A liquid bio-binding mixture is used, which consists of the following components at the following concentrations:

[0595] Reference mixture 20 (R20):

[0596] 48g / L urea

[0597] 44g / L calcium chloride

[0598] 50g / L polyvinyl acetate 20 (solid, granules)

[0599] 4 x 10^8 cells / mL *S. pasteurii*

[0600] Reference mixture 21 (R21):

[0601] 48g / L urea

[0602] 44g / L calcium chloride

[0603] 50g / L polycarbonate (solid, granules)

[0604] 4 x 10^8 cells / mL *S. pasteurii*

[0605] Reference mixture 22 (R22):

[0606] 48g / L urea

[0607] 44g / L calcium chloride

[0608] 50g / L vegetable oil

[0609] 4 x 10^8 cells / mL *S. pasteurii*

[0610] Reference mixture 23 (R23):

[0611] 48g / L urea

[0612] 44g / L calcium chloride

[0613] 12.5 g / L long-chain fatty acids

[0614] 4 x 10^8 cells / mL *S. pasteurii*

[0615] Reference mixture 24 (R24):

[0616] 48g / L urea

[0617] 44g / L calcium chloride

[0618] 50 g / L starch, untreated (solid, powder)

[0619] 4 x 10^8 cells / mL *S. pasteurii*

[0620] Polyvinyl acetate 20 (solid, granules), polycarbonate (solid, granules), rapeseed oil, long-chain fatty acids, and starch were proven to be insoluble in water, non-dispersible in water, and non-emulsifiable in water, and therefore could not be considered viscosity-modifying compounds. Urea in mixtures R20, R21, R22, R23, and R24 was used as a carbonate source.

[0621] Except for rapeseed oil and the strain *S. pasteurii*, all biocementing components of the mixtures of this invention are in solid form. As described, for example, in Cuthbert, MO et al., Ecological Engineering 2012, 41, 32-40 (see section 2.2 on page 33), the bacteria are present in liquid culture in a culture medium known in the art, using 5 g / L yeast extract in the context of this invention. The solid components and the bacteria in the liquid culture are mixed directly before use, and the water-soluble solid components dissolve. Insoluble, non-dispersible, or non-emulsifiable substances are pre-applied uniformly to the top layer of sand.

[0622] Apply the mixture three times repeatedly to the test area. The application rate is always 4 liters per square meter per repeat. Apply the completely dissolved sample using a pipette. After application, smooth the surface with a spatula. The reported measurement is the average of the three replicates, typically within 10% of the determined value.

[0623] Following application of the reference mixture and the bio-binding mixture, incubation was conducted for 28 days during the total observation period, under atmospheric humidity ranging from 20% to 60% and with multiple air changes per day. The lowest temperature observed during this period was 14.2°C, and the highest was 25.2°C.

[0624] The fracture strength of each layer was determined after 1, 2, 3, 4, 10, and 28 days. The fracture strength (maximum force measured) was determined by the following method: This method is based on the standard test method for determining the strength of binders in DIN EN 196-1:2005-05. The fracture strength was measured using a digital (fracture) strength measuring instrument according to the manufacturer's instructions. The specimen was pressed into the sample (up to the fracture point) using a crank-operated test bench, and the applied force was continuously measured. The average fracture strength was calculated through multiple measurements (>3). The average fracture strength was preferably from 0.5 N to 1000 N, more preferably from 1 N to 300 N.

[0625] After determining the fracture strength, the mechanically stressed sample was placed in a wind tunnel and exposed to a wind speed of 12 m / s for 1 minute. The airflow direction impacted the surface at a 12.5° angle. Emission-related weight loss was determined using the formula described in Example 1. This test can serve as a reference for the long-term stability of the sample and its dust suppression.

[0626] To determine the water solubility, water dispersibility, or water emulsification of a substance, the procedure is as follows: To determine the water solubility of solid, paste, and gel substances (e.g., polyvinyl acetate 20, polycarbonate, long-chain fatty acids, and starch), 5 g of the substance is placed in 100 mL of distilled water and stirred at 20°C for 24 hours. The mixture is then filtered (using Homyl 80-120 μm quantitative filter paper). The filter paper is professionally dried and weighed. The determined mass is subtracted from the filter mass, resulting in the mass of the residue, in grams (as defined herein). The difference between 5 g and the mass of the residue (in grams) is divided by 0.1 L to obtain the solubility of the corresponding substance (in grams per liter).

[0627] To determine the water dispersibility of solid, paste, and gel substances, 50 g of the corresponding substance was mixed with 1000 mL of distilled water and incubated at 20 °C. Homogenize in an LC75 dissolver at 15,000 rpm for 5 minutes. The mixture is then transferred to a centrifuge container and centrifuged at 100 g for 2 minutes. The supernatant is decanted, and the precipitate is professionally dried and weighed. The determined mass is the mass of the precipitate after centrifugation (as defined herein). The difference between 50 g and the mass of the precipitate after centrifugation, divided by 1 L, yields the water dispersibility of the substance (as defined herein).

[0628] To determine the water solubility or water emulsification of a liquid substance (e.g., rapeseed oil), the following procedure is used: 5 g of the substance is mixed with 100 g of distilled water and stirred for 24 hours. The mixture is then transferred to a separatory funnel. The mixture is stored in the separatory funnel for 5 minutes. If no phase separation occurs after this time, the mixture is allowed to stand for another 2 hours, preferably another 10 hours. If no phase separation occurs, the substance is considered water-soluble. In this case, the water solubility of the substance is at least 50 g / L. If phase separation occurs, the phase is separated in the separatory funnel and the organic phase is dried with sodium sulfate. The weight of the dried organic phase (mass of the organic phase, in grams, as defined herein) is determined. The difference between 5 g and the mass of the organic phase (in grams) divided by 0.1 L gives the water emulsification of the liquid substance. Water solubility, water dispersibility, and water emulsification are used synonymously in the context of this invention. The limits for the water solubility, water dispersibility, and water emulsification of the compound of component (iii) are defined as 1 g / L, respectively.

[0629] A further preferred separation technique for the dispersed and undispersed fractions is centrifugation. After appropriate drying, the mass of the residue (in grams) can be determined, and the water solubility or water dispersibility can also be determined therefrom.

[0630] result:

[0631] In the preceding Examples 1 to 5, a bio-binding formulation was described that exhibited a synergistic effect with the viscosity-modifying compound and showed reduced emissions-related weight loss.

[0632] After using reference mixtures R20 to R24, it was found that the use of polycarbonate, polyvinyl acetate 20, rapeseed oil, long-chain fatty acids, and insoluble starch did not result in any synergistic effect related to consolidation and emissions reduction. Figure 7 Application of polymers (R15, R16) did not result in a reduction in emission-related weight loss: after four days of reaction and mechanical validation, and one minute of exposure in a 12 m / s wind, the emission-related weight loss of R20 and R21 exceeded 50 wt%. Therefore, this is indistinguishable from the emission-related weight losses of the corresponding reference formulations R3 and R15, and R3 and R16. The lack of a synergistic effect may be due to the non-water solubility of these polymers. In the described assays, the water solubility or water dispersibility of polycarbonate and polyvinyl acetate 20 was less than 1 g / L, respectively (data not shown).

[0633] Applying starch only to the surface resulted in a slight increase in the tensile strength of the layer (11 N), but had no synergistic effect with bio-binding (see [link]). Figure 7 (R19 and R24 in the R19 and R24). The emission-related weight loss is 34 wt%.

[0634] Comparable values ​​were also obtained after 10 days and 28 days (data not shown).

[0635] Comparable effects were also obtained when the bacterial strains were replaced with *L. sphaericus*, *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* at the same cell count per milliliter (data not shown). When *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* were used in the formulation in a similar manner at the same cell count per milliliter, the base composition was further adapted to the requirements of the specific bacterial strains. In this document, those skilled in the art will recognize that, for these non-urea-degrading biocementing bacterial strains, the basal medium must be adjusted similarly to the components listed in Example 2, particularly in terms of suitable metabolic starting materials. The effect of the mixture on emission reduction was comparable to the results illustrated for *S. pasteurii* (data not shown).

[0636] Those skilled in the art will therefore recognize that viscosity-modifying compounds in the sense of this invention must have a certain degree of water solubility and / or water emulsification and / or water dispersibility in order to produce a synergistic effect with bio-binding.

[0637] The removal of the cation source (in this article: calcium source) from the above mixture showed comparable results to dust suppression.

[0638] Example 7: Other applications of the mixture of the invention

[0639] Materials and methods:

[0640] Granulation

[0641] The experiment was conducted in a laboratory granulator. For this purpose, 100g of iron ore (hematite powder) was introduced, and a liquid bio-cementing mixture was used for dust suppression / granulation. This mixture consisted of the following components at the following concentrations:

[0642] Reference mixture 3 (R3):

[0643] 48g / L urea

[0644] 44g / L calcium chloride

[0645] 4 x 10^8 cells / mL *S. pasteurii*

[0646] Mixture 7 (M7):

[0647] 5g / L yeast extract

[0648] 21g / L calcium acetate

[0649] 34.9 g / L calcium chloride

[0650] 46.2 g / L calcium lactate

[0651] 25g / L calcium lignosulfonate

[0652] 4 x 10^8 cells / mL halodurans

[0653] Mixture 8 (M8):

[0654] 36g / L urea

[0655] 35g / L calcium chloride

[0656] 10g / L yeast extract

[0657] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0658] Mixture 9 (M9):

[0659] 27g / L urea

[0660] 17g / L calcium chloride

[0661] 31g / L albumin

[0662] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0663] Mixture 22 (M22):

[0664] 27g / L urea

[0665] 47g / L calcium lignosulfonate

[0666] 12g / L calcium chloride

[0667] 4 x 10^8 cells / mL *S. pasteurii*

[0668] In addition, the mixture contains trace elements and, for example, trace amounts of salt and sugar (less than 1 wt%). Urea in mixtures M8, M9, and M22 is primarily used as a carbonate source. In mixture M7, calcium lactate is used as a carbonate source.

[0669] The calcium lignosulfonate, yeast extract, and albumin in mixtures M7, M8, M9, and M22 are (water-soluble and / or water-dispersible and / or water-emulsifiable) viscosity-modifying compounds.

[0670] Apart from the bacteria, all bio-binding components in the invented mixture are in solid form. The bacteria are present in the form described in Examples 1 to 6. The solid components and the bacteria in the liquid culture are mixed immediately before use, and the solid components dissolve.

[0671] The experiment was also conducted using wood chips to investigate the ability of the bio-bonded mixture to bond the wood chips.

[0672] Spray 20 mL of the corresponding bio-cementing agent mixture onto 100 g of iron ore (hematite powder) and react at a speed of 30 revolutions per minute for 5 minutes.

[0673] Five minutes later, the breaking strength of the obtained particles is determined: for this purpose, a first particle with a similar diameter is selected: the diameter is measured using calipers. The particle diameter is measured at three points on the particle; the deviation of a single measurement is 1 mm. Particles with a diameter corresponding to 11 ± 1 mm are selected. The breaking strength of the particles (maximum force measured) can be determined by the following method: this method is based on the standard test method for determining the strength of binders in DIN EN196-1:2005-05. The breaking strength is measured using a digital (breaking) strength measuring instrument according to the manufacturer's instructions. A cylindrical test plate is mounted on the particle using a crank-operated test bench, and then pressed into the particle (until the breaking point). The applied force is continuously measured. The average breaking strength is calculated from multiple particles (>3). The average breaking strength of the particles is preferably 0.5 N to 500 N, more preferably 1 N to 150 N.

[0674] Evaporation control

[0675] The volume of the experiment in the laboratory is 1000 cm³. 3 The application was carried out in a plastic container. The application area was 29.2 cm² in each case. 2 .

[0676] The soil matrix used in the experiment consisted of silica sand graded 0-2mm. The sand had been washed and dried by the manufacturer and was used directly. Each plastic container used 2200g of silica sand as the soil matrix. The plastic containers were filled.

[0677] As a control, a reference mixture was used, which consisted of the following components at the following concentrations:

[0678] Reference mixture 2 (R2): Water applied.

[0679] A liquid bio-binding mixture is used, which consists of the following components at the following concentrations:

[0680] Reference mixture 3 (R3):

[0681] 48g / L urea

[0682] 44g / L calcium chloride

[0683] 4 x 10^8 cells / mL *S. pasteurii*

[0684] Mixture 11 (M11):

[0685] 48g / L urea

[0686] 44g / L calcium chloride

[0687] 3.9 g / L polyvinyl acetate dispersion

[0688] 4 x 10^8 cells / mL *S. pasteurii*

[0689] Mixture 16 (M16):

[0690] 1.07 g / L ammonium chloride

[0691] 21g / L calcium acetate

[0692] 15g / L L-alanine

[0693] 34.9 g / L calcium chloride

[0694] 0.40 g / L sodium hydroxide

[0695] 1g / L yeast extract

[0696] 46.2 g / L calcium lactate

[0697] 4 x 10^8 cells / mL *A. crystallopoietes*

[0698] Mixture 22 (M22):

[0699] 27g / L urea

[0700] 47g / L calcium lignosulfonate

[0701] 12g / L calcium chloride

[0702] 4 x 10^8 cells / mL *S. pasteurii*

[0703] The mixture also includes trace elements and, for example, trace amounts of salts and sugars (less than 1 wt%). In mixtures R3, M11, and M22, urea is primarily used as a carbonate source. In mixture M16, calcium lactate is used as a carbonate source.

[0704] Yeast extract, L-alanine, polyvinyl acetate dispersion, and calcium lignosulfonate are viscosity-modifying compounds in mixtures M11, M16, and M22.

[0705] Except for strains *A. crystallopoietes*, *L. sphaericus*, and *S. pasteurii*, all components of this bio-gelatinizable mixture are in solid form. The bacteria are present in the culture media described in Examples 1 to 6 in liquid culture form. The solid components and the bacteria in the liquid culture are mixed immediately before use, dissolving the solid components.

[0706] Before applying the respective mixtures, moisten the sand with water so that the sand is completely soaked when the mixture is subsequently applied. Then, apply the respective mixtures three times repeatedly to the test area. The application rate is always 10 liters per square meter per repeat. Apply the completely dissolved sample using a pipette. After application, smooth the surface with a spatula. The reported measurements are the average of the three replicates, typically within 10% of the determined value.

[0707] The applied mass of water was determined by gravity. For this purpose, the mass of the sand-filled sample container was measured before and after the application of water and the corresponding biocementing mixture (mass before application and mass after application, both defined herein). The difference between the mass before and after application, minus the solids contained in the corresponding biocementing mixture (see M11, M16, M22), yielded the amount of water applied (defined herein). The solids present in the corresponding biocementing mixture were given by multiplying the corresponding solids concentration by the corresponding application volume. The sum of the mass before application and the solids contained in the corresponding biocementing mixture was the total solids mass of the beaker (defined herein).

[0708] Following application of the reference mixture and the bio-binding mixture, incubation was performed for 168 days at atmospheric humidity ranging from 20% to 60% with multiple air changes per day throughout the observation period. During this period, the lowest temperature observed was 14.2°C, and the highest was 25.2°C; all mixtures were exposed to identical external conditions. The sample container mass (sample mass) was measured and recorded at different time points. 第xy天 ).

[0709] The relative soil moisture on the corresponding measurement day is determined using the following formula, expressed in % (day xy):

[0710] Relative soil moisture 第xy天 =[(sample mass)] 第xy天 [(Total solid mass of beaker) / Amount of water applied] * 100

[0711] The experiment was also conducted using wood chips, mine tailings, and rural soil. For this purpose, the top 5 cm of the sand layer was replaced with wood chips, mine tailings, or rural soil, and as described above, it was treated as a soil matrix using mixtures R3, M11, M16, and M22. The total solid mass was adjusted based on the weight of the wood chips, mine tailings, and rural soil, respectively.

[0712] Stain removal

[0713] A liquid bio-binding mixture is used, which consists of the following components at the following concentrations:

[0714] Reference mixture 9 (R9)

[0715] 25g / L polyvinyl alcohol

[0716] Reference Mixture 25 (R25):

[0717] 48g / L urea

[0718] 4 x 10^8 cells / mL *S. pasteurii*

[0719] Mixture 20 (M20):

[0720] 27g / L urea

[0721] 47g / L calcium lignosulfonate

[0722] 4 x 10^8 cells / mL *S. pasteurii*

[0723] Mixture 29 (M29):

[0724] 1g / L yeast extract

[0725] 25g / L calcium lignosulfonate

[0726] 21g / L sodium acetate

[0727] 46.2 g / L sodium lactate

[0728] 4 x 10^8 cells / mL Bacillus cohnii

[0729] Mixture 30 (M30):

[0730] 48g / L urea

[0731] 50g / L humic acid

[0732] 4 x 10^8 cells / mL *S. pasteurii*

[0733] Mixture 31 (M31):

[0734] 27g / L urea

[0735] 47g / L sodium lignosulfonate

[0736] 4 x 10^8 cells / mL *S. pasteurii*

[0737] Mixture 32 (M32):

[0738] 48g / L urea

[0739] 25g / L polyvinyl alcohol

[0740] 4 x 10^8 cells / mL of spherical lysine-containing Bacillus (L. sphaericus)

[0741] In addition, mixtures R25, M20, M29, M30, M31, and M32 contain trace elements and, for example, trace amounts of salts and sugars (less than 1 wt%). Urea in mixtures M20, M30, M31, and M32 is primarily used as a carbonate source; sodium acetate and sodium lactate in mixture M29 are primarily used as carbonate sources. These mixtures optionally contain one of the following metal salts (0.1 M): nickel(II), ferric(III), or copper(II). If ferric(III) is used, hydrochloric acid (0.1 M) is also present. Each metal salt is mixed with each mixture. The names used are as follows: metal salt + corresponding mixture. For metal salts, the following names are used: ferric(III) = FeCl3, nickel(II) = NiCl2, copper(II) = CuCl2. For example, a mixture that includes copper(II) in addition to reagent 20 is listed as CuCl2 + M2O (see [link to reagent 20]). Figure 8 Each metal salt solution was also treated accordingly without the addition of the corresponding mixture.

[0742] All components, including the corresponding bacteria, are in solid form. In the case of powdered bacteria, the powder is a professionally dried powder. Except for the corresponding bacterial powder, all components are mixed directly before use, and the solid components dissolve. Once the components are completely dissolved, the corresponding bacterial powder is added and dissolved.

[0743] The mixture was combined with bacterial powder and stirred for 5 minutes, then reacted for 24 hours. The resulting precipitate was then separated by centrifugation (3000 g, 10 minutes) and decanted. The mass of the wet heavy metal-containing precipitate was determined – the wet mass of the heavy metal-containing precipitate (as defined herein). The wet heavy metal-containing precipitate was then dried in a nitrogen stream, and the mass of the heavy metal-containing precipitate (as defined herein) was determined. The presence of the corresponding heavy metal ions was quantitatively confirmed by atomic spectroscopy. As a control, the corresponding mixtures were prepared in the absence of the corresponding metal salts (M20, M29, M30, M31, and M32) and treated according to the exact same procedure (5 minutes stirring, 24 hours reaction, centrifugation, decantation, drying). The mass of the precipitate after decantation was the wet mass of the control precipitate (as defined herein). After drying, the mass of the control precipitate (as defined herein) was determined. The presence of heavy metal ions in the corresponding supernatant was also investigated by absorption spectroscopy and / or atomic spectroscopy. In this case, appropriate wavelengths with suitable sensitivity were used. The concentration of the corresponding heavy metal ions obtained from this analysis is the residual heavy metal ion concentration (as defined in this paper). The residual heavy metal ion concentration divided by 0.1 mol / L and multiplied by 100 is the percentage of residual heavy metal ions in the supernatant (as defined in this paper). Similarly, the corresponding metal salt solution is stirred for 5 minutes, incubated for 24 hours, centrifuged at 3000g for 10 minutes, and then the residual heavy metal ion concentration is determined.

[0744] result:

[0745] Further unexpected observations were obtained when the invented bio-cementing mixture was tested on various matrices, which will be elaborated further below. Therefore, further potential applications arise, which are further illustrated in this embodiment:

[0746] Granulation

[0747] When the invented formulation was applied to mobile ion mineral samples, small agglomerates or particles were observed to form after application to prevent dust formation. This observation was made to form particles in laboratory granulation plates using bio-binding mixtures M7, M8, M9, and M22.

[0748] In addition to reducing emissions during the production process, these particles also exhibit greater strength than the reference formulation (R3). Figure 8 The image above shows particles produced using different reagents M7, M8, and M9 (from left to right) in each case.

[0749] For various reagents, the breaking strength of the particles is as follows: M7 = 28 N, M8 = 29 N, M9 = 30 N, M22 = 27 N, thus higher than R3. Particles produced using R3 have a breaking strength of 3 N 5 minutes after production. It has been shown that the particles produced using reference mixture 3 are difficult to process because the particles produced using R3 are very easily broken. This is likely due to the lack of viscosity-modifying compounds. Therefore, viscosity-modifying substances also allow for the production of bio-binders that can be used for granulation.

[0750] The mixture according to the invention can also bind and aggregate wood chips.

[0751] Similar results in emission reduction were also found when a mixture in which all components (including bacteria) were in powder form was used. For this purpose, all powdered components were mixed and the aforementioned amount of water (data not shown) was added in a laboratory granulator. Comparable effects were also obtained when the bacterial strains were replaced with *L. sphaericus*, *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* at the same cell count per milliliter (data not shown). When *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* were used in a similar manner at the same cell count per milliliter in the formulation, the basic components were further adapted to the requirements of specific bacterial strains. In this paper, those skilled in the art will recognize that, in the case of these non-urine-soluble biogelatinizing bacterial strains, the basal culture medium must be adjusted in accordance with the components listed in Example 2, particularly regarding suitable metabolic starting materials. In this case, it was found that all bacterial strains achieved efficient granulation (data not shown).

[0752] Surprisingly, it turned out that some matrices dried more slowly in the laboratory granulator after the formation of the bio-binding agent, thus allowing for a deeper investigation into the effect of the bio-binding mixture on evaporation.

[0753] Evaporation control

[0754] The effective cambium layer reduced the drying rate of the sand. This is evident from the higher relative soil moisture content of samples from mixtures M11, M16, and M22 compared to the applied water (R2). Figure 8(middle). Compared with the application of the reference formulation R3, which is capable of biocementing, the relative soil moisture of mixtures M11, M16, and M22 was significantly higher. This is because the resulting biocementing layer has a viscosity barrier to the downward-flowing water. It can be hypothesized that the porosity of this layer is altered in some way due to the presence of viscosity-modifying substances, thereby allowing water to evaporate more slowly.

[0755] The altered porosity may also be relevant to other applications where porosity plays a role. This is especially true in applications such as insulating materials, catalyst beds, and / or battery materials. Due to the reduced porosity, the material is also suitable for use as a sealing material.

[0756] Similar results in emission reduction were also found when a mixture in which all components (including bacteria) were in powder form was used. For this purpose, all powdered components were mixed and incorporated into the top layer. The corresponding liquid volume was then applied (data not shown). Comparable effects were also obtained when the bacterial strains were replaced with *L. sphaericus*, *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* at the same cell count per milliliter (data not shown). When *B. cohnii*, *B. halodurans*, *B. pseudofirmus*, and *A. crystalloietes* were used in a similar manner at the same cell count per milliliter in the formulation, the basic components were further adapted to the requirements of specific bacterial strains. In this paper, those skilled in the art will recognize that, in the case of these non-urine-soluble bio-gelling bacterial strains, the basal culture medium must be adjusted in accordance with the components listed in Example 2, particularly regarding suitable metabolic starting materials. In this case, it was found that all bacterial strains effectively reduced evaporation (data not shown).

[0757] When using various mine tailings, soils with high copper(II), iron(II), iron(III), and nickel(II) ion loadings were found to form layers significantly faster. Similar results were obtained when tailings / sand stratification was used, as with pure sand samples (data not shown). Similarly, mixtures with viscosity modifiers dried more slowly in this study.

[0758] Stain removal

[0759] Those skilled in the art will recognize that, for example, carbonate ions produced by bacteria from urea can be used to precipitate metal ions (Phillips et al., Engineering applications of urea decomposition biomineralization: a review, Biodeposition, 2013, Vol. 29, No. 6, 715-733). This may explain the observation in the previous embodiment (Example 7, evaporation control) that layer formation began more rapidly when heavy metal-loaded soil was used. Therefore, tests were conducted to determine whether viscosity modifiers were also suitable for improving heavy metal ion precipitation.

[0760] In each mixture used, the mass of the control precipitate was less than the mass of the heavy metal-containing precipitate. This indicates that the mixtures according to the invention can bind and precipitate heavy metal ions. The presence of the corresponding metal salts was confirmed by atomic spectroscopy.

[0761] When ferric chloride (III) was precipitated using R9, R25, and R32, the wet mass of the heavy metal-containing precipitate was FeCl3+R9 = 0.00 g, FeCl3+R20 = 0.75 g, and FeCl3+M32 = 12.4 g (before drying). In the case of FeCl3+M32, bacterial activity led to the formation of a large-volume gel. This gel was difficult to dry in a nitrogen stream. The mass of the heavy metal-containing precipitate was 0.00 g for FeCl3+R9, 0.05 g for FeCl3+R25, and 6.53 g for FeCl3+M32 (this paper assumes that the gelling properties lead to incomplete drying). The residual heavy metal ion content of FeCl3+M32 was 50% lower than that of FeCl3+R25.

[0762] When using M2O to precipitate FeCl3, NiCl2, and CuCl2, it was found that the addition of viscosity-modifying compounds led to an increase in metal salt precipitation: the wet mass of M2O precipitation was 0.02 g. The wet mass of heavy metal-containing precipitates from FeCl3 + M2O was 0.40 g, NiCl2 + M2O was 1.44 g, and CuCl2 + M2O was 0.24 g. The residual heavy metal ion content in the supernatant was as follows: Figure 8 As shown below.

[0763] Surprisingly, the presence of viscosity modifiers also showed an increase in precipitation in solution. The use of mixtures not explicitly mentioned in the results demonstrated comparable results related to precipitation efficiency (data not shown). Bacteria from the liquid culture media used in Examples 1 through 6 also exhibited very good precipitation efficiency (data not shown).

[0764] A key characteristic of viscosity-modifying compounds is their synergistic effect with bio-binding agents, resulting in high-viscosity, low-emission bio-binding agents. Unexpectedly, this also occurs in solution, leading to particularly efficient precipitation of heavy metal ions. This is surprising, especially since polymers have a particular tendency to bind polyvalent ions, particularly divalent metal cations such as Ca(II), Cu(II), Mg(II), and Ni(II), to disperse them in solution, thereby increasing their solubility. Based on this affinity for divalent metal cations, it is expected that, especially if the aqueous and / or water-dispersible viscosity-modifying compounds are polymers, they will stabilize polyvalent metal cations and their aggregates and agglomerates in solution, leading to reduced metal ion precipitation efficiency (see Tadros TF 2016, Nanodispersions, ISBN-978-3-11-029033-2, especially section p. 25ff Stereostabilization).

[0765] The removal of the cation source (in this paper: calcium source) from the above mixture showed results comparable to those studied.

Claims

1. Use of a mixture suitable for bio-cementing, said mixture having the following components: (i) at least one organism capable of forming carbonates or capable of inducing and / or catalyzing carbonate formation, and / or at least one enzyme capable of forming carbonates or capable of inducing and / or catalyzing carbonate formation. (ii) at least one substance used to form carbonates, (iii) at least one water-soluble, water-dispersible, and / or water-emulsifiable viscosity-modifying compound, (iv) Optionally: at least one cation source; (v) Optionally: at least one adjuvant; The purpose of the mixture is to reduce dust formation and / or erosion. Used for granulation. Used to reduce evaporation As a component of sealing or insulating materials, or as a sealing or insulating material, and / or Used for heavy metal precipitation. The viscosity-modifying compound mentioned above is selected from the group consisting of the following substances: Compounds with calcium affinity; and Compounds with carbonate affinity; Furthermore, the compounds having calcium affinity and carbonate affinity mentioned therein are selected from the group consisting of: (iii-1) Polymers, selected from the group consisting of: Lignin and its derivatives Polyhydroxybutyrate, polylactide, polyacrylic acid, polyacrylate, polyvinyl alcohol, polyvinyl acetate, polyvinylpyrrolidone, polystyrene, styrene-butadiene, styrene-acrylate, ethylene-vinyl acetate; isocyanate, polyamino acid; Cellulose and its derivatives, starch and its derivatives, humic acid and its derivatives; Chitosan and its derivatives, cyclodextrin and its derivatives, dextrin and its derivatives Protein sources and / or peptides containing at least one of the following amino acids: alanine, glycine, lysine, asparagine, glutamine, and glutamic acid; Yeast extract; (iii-2) Sugars, wherein the sugars include lactose, sucrose, glucose, glucosamine, fructose, inulin and combinations thereof, or constitute the sugars thereof; (iii-3) Carboxylic acids, selected from the group consisting of fruit acids, short-chain and medium-chain fatty acids, and lactic acid, and in each case their salts and their esters; (iii-4) Inorganic binders, minerals and salts selected from the group consisting of CEM I, CEM II, CEM III, CEM IV, CEM V, CEMVI, alumina binders, magnesium oxide binders, gypsum, sodium silicate, potassium silicate, calcium carbonate, aluminum hydroxide, calcium sulfate, calcium hydroxide, magnesium sulfate, microsilica and kaolin. (iii-5) Amino acids selected from the group consisting of alanine, glycine, lysine, glutamine, and glutamic acid, and in each case their salts, esters, and amides; and (iii-6) Gum arabic, xanthan gum, alginate and agar.

2. The use according to claim 1, wherein in (iii-3), the carboxylic acid is selected from citric acid, formic acid, propionic acid, acetic acid, and in each case its salts and esters.

3. The use according to claim 1, wherein in (iii-5), the salt is a carboxylate.

4. The use according to claim 1, wherein the effect produced by said components (i), (ii) and (iii) is greater than the sum of the effects produced by said components (i) and (ii) and the effect produced by said component (iii).

5. The use according to claim 4, wherein the effect is a dust reduction effect.

6. The use according to claim 1, wherein the component (iii) is present in an amount of at least 0.5 wt% based on the total mass of the components (i), (ii) and (iii), and / or Based on the total mass of components (i), (ii) and (iii), component (iii) is present in an amount of up to 85 wt%.

7. The use according to claim 1, wherein the component (iii) is selected from the group consisting of: lignin sulfonates, humic acid and its salts, lignin sulfates, gum arabic, xanthan gum, alginate, and agar. Casein, albumin, peptone, caseinate, calcium caseinate, milk powder Polymethacrylate, poly(2-hydroxyethyl methacrylate). Corn extract, lactose mother liquor, protein lysate, molasses, and protein waste.

8. The use according to claim 7, wherein the protein waste is selected from the group consisting of protein waste derived from yeast, protein waste derived from meat, protein waste derived from fruit, protein waste derived from vegetables, and protein waste derived from the dairy industry.

9. The use according to claim 1, wherein the component (iii) is selected from the group consisting of: Calcium lignosulfonate, sodium lignosulfonate, potassium lignosulfonate, magnesium lignosulfonate, ammonium lignosulfonate, yeast extract, albumin, starch ether, alanine, lysine, styrene-acrylate dispersion, magnesium sulfate, polyvinyl acetate dispersion, styrene-butadiene dispersion, humic acid, sodium silicate, potassium silicate, starch ester, and combinations thereof.

10. The use according to claim 9, wherein the component (iii) is selected from calcium lignosulfonate.

11. The use according to claim 1, wherein the component (ii) is selected from the group consisting of: Urea and its salts; organic acids and their salts and esters; peptides containing non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine and / or glutamic acid; amino acids and their salts and esters; plant and animal composite substrates; industrial residue streams; anaerobic substances.

12. The use according to claim 1, wherein the component (ii) is selected from the group consisting of: lactic acid and its salts and esters; gluconic acid and its salts and esters; acetic acid and its salts and esters; formic acid and its salts and esters; propionic acid and its salts and esters; butyric acid and its salts and esters; valeric acid and its salts and esters; maleic acid and its salts and esters; succinic acid and its salts and esters; pyruvic acid and its salts and esters; acetoacetic acid and its salts and esters; acetopropionic acid and its salts and esters; oxaloacetic acid and its salts and esters; and fruit acids and their salts and esters.

13. The use according to claim 12, wherein the fruit acid is citric acid.

14. The use according to claim 1, wherein the component (ii) is selected from the group consisting of: malic acid and its salts and esters; citric acid and its salts and esters; fumaric acid and its salts and esters; gluconic acid and its salts and esters; glycolic acid and its salts and esters; mandelic acid and its salts and esters; oxalic acid and its salts and esters; salicylic acid and its salts and esters; α-hydroxyoctanoic acid and its salts and esters; and tartaric acid and its salts and esters.

15. The use according to claim 11, wherein the amino acid is selected from the group consisting of: non-proteinogenic amino acids, asparagine, alanine, glycine, lysine, glutamine, and glutamic acid.

16. The use according to claim 11, wherein the salt is a carboxylate.

17. The use according to claim 11, wherein the plant and animal composite matrix is ​​selected from peptone, yeast extract, meat extract, nutrient broth and casein amino acids.

18. The use according to claim 11, wherein the industrial residue stream is selected from corn extract, lactose mother liquor, and protein lysate.

19. The use according to claim 11, wherein the industrial residue stream is derived from peas, meat, potatoes, or tomatoes.

20. The use according to claim 11, wherein the anaerobic substance is carbon dioxide and methane.

21. The use according to claim 1, wherein the mixture is present in liquid form, gel, paste or powder.

22. The use according to claim 1, wherein the component (i) is selected from the group consisting of microorganisms.

23. The use according to claim 22, wherein the microorganism is selected from the group consisting of microorganisms of Firmicutes, Proteobacteria, Actinobacteria, and Cyanobacteria; and aerobic bacteria, anaerobic bacteria, and facultative anaerobic bacteria.

24. The use according to claim 1, wherein the component (i) is selected from metabolic enzymes.

25. The use according to claim 1, wherein the component (i) is selected from the group consisting of urease, asparaginase, and carbonic anhydrase.

26. The use according to claim 1, wherein the component (v) is selected from the group consisting of: Natural and chemical herbicides; fungicides, molluscicides; insecticides; hydrophobic agents and wax emulsions; stabilizers, dispersants; emulsifying agents, surfactants; amines; thixotropic agents; propellants; free-flowing agents, seed crystals and crystallization modifiers; complexing agents; minerals and trace elements; salts; rocks; rubber debris, rubber granules and other thermoplastic elastomers; aggregates; spores; plants and their parts; fertilizers; bacteria capable of forming polymers; and modified bio-binding substances.

27. The use according to claim 26, wherein the amine is ethanolamine and the plant and a portion thereof are plant seeds.

28. The use according to claim 26, wherein the surfactant is a cationic, anionic, or uncharged surfactant.

29. The use according to claim 26, wherein the complexing agent is selected from phosphonates, phosphates, polyphosphates, and fatty acids.

30. The use according to claim 26, wherein the salt is selected from halides, silicates, phosphates and sulfates.

31. The use according to claim 26, wherein the rock is selected from pumice, sand, gravel and slate powder.

32. The use according to claim 26, wherein the rubber debris, rubber particles and other thermoplastic elastomers are derived from the tire industry.

33. The use according to claim 26, wherein the aggregate is selected from amorphous and crystalline aggregates.

34. The use according to claim 26, wherein the aggregate is selected from hydraulic, non-hydraulic and hardening materials.

35. The use according to claim 27, wherein the plant seeds are derived from monocotyledonous and dicotyledonous plants.

36. The use according to claim 26, wherein the spores are bryozoans.

37. The use according to claim 26, wherein the plant and its parts are roots, bulbs, and wood.

38. The use according to claim 26, wherein the plant and a portion thereof are wood chips.

39. The use according to claim 1, wherein the component (i) is present in the matrix to be treated by the mixture and / or a portion of the matrix, and wherein components (ii) and (iii), and optionally components (iv) and / or (v), are applied separately from component (i), or The component (i) is separated from the matrix to be treated by the mixture, cultured in situ, and then either in combination with components (ii) and (iii) and optionally with components (iv) and / or (v), or separately from components (ii) and (iii) and optionally with components (iv) and / or (v), and introduced onto and / or into the matrix to be treated.

40. The use according to claim 39, wherein the substrate is soil.

41. Methods for reducing dust formation and / or erosion, including the following steps: (a) Identify the substrate to be treated and reduce dust formation and / or erosion on the substrate. (b) Provide a mixture or all components of a mixture as defined in any one of claims 1 to 40, (c) Apply the mixture or all components of the mixture provided in step (b) to the substrate to be treated in an amount sufficient to achieve bio-binding, and (d) Allows the formation of a bio-binding agent layer, thereby reducing dust formation and / or erosion on the substrate.

42. Methods for reducing dust formation and / or erosion, including the following steps: (a) Identify the substrate to be treated, and reduce dust formation and / or erosion within the substrate. (b) Provide a mixture or all components of a mixture as defined in any one of claims 1 to 40, (c) Apply the mixture or all components of the mixture provided in step (b) to the matrix to be treated in an amount sufficient to achieve bio-binding, and (d) Allows the formation of a bio-binding agent layer, thereby reducing dust formation and / or erosion within the matrix.

43. The method according to any one of claims 41 to 42, wherein the matrix is ​​selected from the group consisting of organic and inorganic materials, their derivatives and mixtures, and combinations thereof.

44. The method of claim 43, wherein the matrix is ​​selected from biological and / or anthropogenic materials.

45. The method of claim 43, wherein the matrix is ​​selected from metamorphic rocks, sedimentary rocks, and igneous rocks.

46. ​​The method according to any one of claims 41 to 42, wherein step (c) is performed once or repeatedly, and the total amount of said component (iii) applied is at least 20 g based on an application area of ​​one square meter, and / or Based on an application area of ​​one square meter, the total amount of said component (iii) applied is at most 2000 g, and / or The component (i) provided in step (b) is a native organism present in the matrix to be treated.

47. A biocementable mixture as defined in claim 1, wherein component (iii) is selected from the group consisting of: Albumin; starch ether; alanine; lysine; styrene-acrylate; ethylene-vinyl acetate; polyvinyl alcohol; magnesium sulfate; polyvinyl acetate; styrene-butadiene; humic acid and combinations thereof Component (ii) is selected from the group consisting of the following substances: Urea and its salts; organic acids and their salts and esters; peptides containing asparagine, alanine, glycine, lysine, glutamine and / or glutamic acid; amino acids and their salts and esters; plant and animal composite substrates; industrial residue streams; anaerobic substances.

48. The bio-cementable mixture according to claim 47, wherein the styrene-acrylate is a styrene-acrylate dispersion.

49. The bio-cementable mixture according to claim 47, wherein the ethylene-vinyl acetate is an ethylene-vinyl acetate dispersion.

50. The bio-binding mixture according to claim 47, wherein the polyvinyl acetate is a polyvinyl acetate dispersion.

51. The bio-cementable mixture according to claim 47, wherein the styrene-butadiene is a styrene-butadiene dispersion.

52. The bio-cementing mixture according to claim 47, wherein the component (ii) is selected from the group consisting of: lactic acid and its salts and esters; gluconic acid and its salts and esters; acetic acid and its salts and esters; formic acid and its salts and esters; propionic acid and its salts and esters; butyric acid and its salts and esters; valeric acid and its salts and esters.

53. The bio-cementable mixture according to any one of claims 47 to 52, wherein the amino acid is selected from asparagine, alanine, glycine, lysine, glutamine, and glutamic acid.

54. The bio-binding mixture according to any one of claims 47 to 52, wherein the plant and animal composite matrix is ​​peptone, yeast extract, meat extract, nutrient broth, and casein amino acids.

55. The bio-cementable mixture according to any one of claims 47 to 52, wherein the industrial residue stream is corn extract, lactose mother liquor, or protein lysate.

56. The bio-binding mixture according to any one of claims 47 to 52, wherein the industrial residue stream is derived from peas, meat, potatoes, or tomatoes.

57. The biocementable mixture according to any one of claims 47 to 52, wherein the anaerobic substance is carbon dioxide and methane.

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