ECOLOGICAL RELEASE OF ELEMENTS AND DEGRADATION OF ORGANICS USING HETEROTROPHIC MICROORGANISMS OUT OF MULTIPLE CARRIER MATERIALS

Abstract

The present invention relates to a process of releasing metallics, metalloids and/or non-metallics and/or degradating organics by bioleaching out of a carrier material, such as minerals, rocks, wastes, soil, and sediments. Furthermore, the present invention relates to a leached formulation, obtainable form said process, and to the use of said leached formulation as a biofertilizer or biostimulant or biopestizide.

Claims

1-15. (canceled)

16: A process of releasing metallics, metalloids and/or non-metallics and/or degradating organics out of a carrier material, the process comprising: a. providing and multiplying a solution of natural consortia of heterotrophic bacteria; b. providing a carrier material in which at least one of metallic, metalloid, nonmetallic and organic components is found, c. inoculating the carrier material with the solution of natural consortia provided in step a; and d. bioleaching the at least one of metallic, metalloid, non-metallic and organic components out of the carrier material.

17: The process according to claim 16, wherein the carrier material is selected from the group consisting of minerals, rocks, wastes, soil, and sediments.

18: The process of claim 16, wherein the solution used for bioleaching is a mixture of at least two of Lactobacillus, Leuconostoc, Lactococcus, Bacillus, Clostridium, Bifidobacterium, Streptococcus, Acetobacter, Pseudomonas, Enterobacter, Oenococcus, Pediococcus, Enterococcus, Citrobacter, Acinetobacter, Azotobacter and Arthrobacter, whereby one genus of them predominates in an amount of at least 10.sup.3 CFU/g in solid sample.

19: The process of claim 17, wherein the solution used for bioleaching is a mixture of at least two of Lactobacillus, Leuconostoc, Lactococcus, Bacillus, Clostridium, Bifidobacterium, Streptococcus, Acetobacter, Pseudomonas, Enterobacter, Oenococcus, Pediococcus, Enterococcus, Citrobacter, Acinetobacter, Azotobacter and Arthrobacter, whereby one genus of them predominates in an amount of at least 10.sup.3 CFU/g in solid sample.

20: The process of claim 18, characterized in that the genus Lactobacillus, Leuconostoc, Lactococcus, Bacillus, Clostridium, Bifidobacterium, Streptococcus, Acetobacter, Pseudomonas, Enterobacter, Oenococcus, Pediococcus, Enterococcus, Citrobacter, Acinetobacter, Azothobacter or Arthrobacter used in the bioleaching step originates from natural sources from the manifestation of weathering processes of minerals and organic materials, such as grass, leaves, wood, and compost, and obtaining biogenic silicate elements.

21: The process of claim 16, wherein bioleaching is carried out at a temperature of at least 5° C.

22: The process of claim 16, wherein the bioleaching is carried out in nonsterile conditions ex-situ or in-situ.

23: The process of claim 16, wherein the process is carried out under industrial conditions in unsterile basins and heaps.

24: The process of claim 16, wherein the bioleaching is carried out under weathering or non-weathering conditions, such as a closed hall.

25: The process of claim 24, wherein the carrier material contains organic components, and said organic components produce by fermentation during bioleaching at least one of acetic acid, butyric acid, pyruvic acid, lactic acid, formic acid, acetoacetic acid, succinic acid, valeric acid, fumaric acid, malic acid, glycolic acid, citric acid, oxalic acid, formic acid and/or propionic acid

26: The process of claim 16, wherein the volume of the solution of natural consortia of heterotrophic bacteria is at least 0.01% of the volume of the carrier material.

27: The process of claim 16, wherein the solution of natural consortia of heterotrophic bacteria has a pH value of between 3.5 and 9.5.

28: The process of claim 16, wherein the carrier material in which at least one of metallic, metalloid, non-metallic and organic components is found does not contain organic material, and the solution of natural consortia of heterotrophic bacteria comprises organic components selected from the group consisting of molasses, sugar, glucose, sucrose, dextrose, fucose, galactose, fructose, mannitol, maltose, glycerol. cellulose, compost, sewage sludge and digestate.

29: The process of claim 17, wherein the carrier material in which at least one of metallic, metalloid, non-metallic and organic components is found does not contain organic material, and the solution of natural consortia of heterotrophic bacteria comprises organic components selected from the group consisting of molasses, sugar, glucose, sucrose, dextrose, fucose, galactose, fructose, mannitol, maltose, glycerol. cellulose, compost, sewage sludge and digestate.

30: The process of claim 18, wherein the carrier material in which at least one of metallic, metalloid, non-metallic and organic components is found does not contain organic material, and the solution of natural consortia of heterotrophic bacteria comprises organic components selected from the group consisting of molasses, sugar, glucose, sucrose, dextrose, fucose, galactose, fructose, mannitol, maltose, glycerol. cellulose, compost, sewage sludge and digestate.

31: The process of claim 16, wherein the solution of natural consortia further comprises at least one chelate, selected from the group consisting of ethylene diamine disuccinic acid, tri-sodium salt of (S,S)-ethylene diamine disuccinic acid, ethylene diamine tetraacetic acid (EDTA), calcium disodium EDTA, diammonium EDTA, dipotassium EDTA, disodium EDTA, triethanolamine salt of EDTA, tripotassium EDTA, trisodium EDTA, trisodium EDTA, tetrasodium EDTA, diethylene triaminepentaacetic acid, N-(hydroxyethyl)-ethylene diaminetetraacetic acid, L-glutamic acid-N,N-diacetic acid, nitrilotriacetic acid, methyl glycinediacetic acid, hydroxy ethyliminodiacetic acid, iminodisuccinic acid, ethylenediamine-N,N′-diglutaric acid, ethylenediamine-N,N′-dimalonic acid, 3-hydroxy-2,2-iminodisuccinic acid, pyridine-2,6-dicarboxylic acid, nitrilotrimethylenephosphonic acid, glucoheptonate, diethylenetriamine penta(methylene phosphonic acid), hydroxyethylidenediphosphonic acid, and nitrilotrimethylenephosphonic acid.

32: The process of claim 18, wherein the solution of natural consortia further comprises at least one chelate, selected from the group consisting of ethylene diamine disuccinic acid, tri-sodium salt of (S,S)-ethylene diamine disuccinic acid, ethylene diamine tetraacetic acid (EDTA), calcium disodium EDTA, diammonium EDTA, dipotassium EDTA, disodium EDTA, triethanolamine salt of EDTA, tripotassium EDTA, trisodium EDTA, trisodium EDTA, tetrasodium EDTA, diethylene triaminepentaacetic acid, N-(hydroxyethyl)-ethylene diaminetetraacetic acid, L-glutamic acid-N,N-diacetic acid, nitrilotriacetic acid, methyl glycinediacetic acid, hydroxy ethyliminodiacetic acid, iminodisuccinic acid, ethylenediamine-N, N′-diglutaric acid, ethylenediamine-N,N′-dimalonic acid, 3-hydroxy-2,2-iminodisuccinic acid, pyridine-2,6-dicarboxylic acid, nitrilotrimethylenephosphonic acid, glucoheptonate, diethylenetriamine penta(methylene phosphonic acid), hydroxyethylidenediphosphonic acid, and nitrilotrimethylenephosphonic acid.

33: The process of claim 25, wherein the solution of natural consortia further comprises at least one chelate, selected from the group consisting of ethylene diamine disuccinic acid, tri-sodium salt of (S,S)-ethylene diamine disuccinic acid, ethylene diamine tetraacetic acid (EDTA), calcium disodium EDTA, diammonium EDTA, dipotassium EDTA, disodium EDTA, triethanolamine salt of EDTA, tripotassium EDTA, trisodium EDTA, trisodium EDTA, tetrasodium EDTA, diethylene triaminepentaacetic acid, N-(hydroxyethyl)-ethylene diaminetetraacetic acid, L-glutamic acid-N,N-diacetic acid, nitrilotriacetic acid, methyl glycinediacetic acid, hydroxy ethyliminodiacetic acid, iminodisuccinic acid, ethylenediamine-N, N′-diglutaric acid, ethylenediamine-N,N′-dimalonic acid, 3-hydroxy-2,2-iminodisuccinic acid, pyridine-2,6-dicarboxylic acid, nitrilotrimethylenephosphonic acid, glucoheptonate, diethylenetriamine penta(methylene phosphonic acid), hydroxyethylidenediphosphonic acid, and nitrilotrimethylenephosphonic acid.

34: A leached carrier material, obtainable form the process according to claim 16.

35: A process of promoting plant growth comprising, applying to or near a plant, the leached carrier material of claim 33.

Description

EXAMPLE 1

[0065] 300 tons of silica sand product MAP1 of Termit d.d., Drtija 51, 1251 Moravc̆e/Slovenia was used.

[0066] The process according to the present invention was carried out in 3 basins excavated in sand, covered by clay layer and additional folia to secure impermeability. The basins have been filled with 300 tons of the silica sand product MAP1 first. 300 m.sup.3 of water from nearby creek used in sand processing was added. In parallel, a consortium of allochthonous bacteria were multiplicated up to an amount of 0.01% related to the raw material to be treated. The active mixture of isolated heterotrophic bacteria were belonging to the genus Lactobacillus, Leuconostoc, Lactococcus, Bacillus, Clostridium, Bifidobacterium, Streptococcus, Acetobacter, Enterobacter, Oenococcus, Pediococcus, Enterococcus, Citrobacter and Pseudomonas. They were identified by means of the Becton-Dickinson microbiology system (Becton Dickinson, Cockeysville, USA).

[0067] After 4 days of bioleaching, the bacteria were inoculated in the basins. As next step all nutrition components for the bacteria were added (with a total amount of 7.2 t). The nutrition components were as follows:

TABLE-US-00001 element content (mg/l) N 392.2 P 28.8 K 211 Ca 49.1 Cl 295.8 Mg 49.3 Na 674.9 S 516.5 Na.sub.4EDTA 760 Sugar 30000

[0068] Outside temperature along the whole procedure was in average 17° C. (the highest 30° C., the lowest 6° C.). The pH value decreased from 7 to 4 after 10 days of bioleaching. The production of gases with bubble formation proved evidence of the fermentation of organic source and suitable bioleaching. As the water supply was available only during weekends (due to running processing in the plant), the leachate was pumped out and replaced by new water and nutrition after 14 days. The process was conducted under weathering and non-sterile conditions for 1 month. The iron concentration in the leachate after the first exchange of media was 350 mg/l and on 150 mg/l after the second removal of leachate. All iron oxides originally covering every quartz grain (responsible for the brown colour of the sand) were dissolved during the bioleaching process decreasing the Fe.sub.2O.sub.3 content from 0.13 wt % to 0.06 wt % (53.8% removal). For separating the remaining black resistant magnetic minerals, a combination with magnetic separation was used which decreased the iron content bellow 0.03% (300 ppm), fulfilling the request of the interested end-customer (a flint glass producer).

EXAMPLE 2

[0069] To compare the effectivity of the mixed bacteria consortia, a smaller test was conducted at the same conditions at Termit d.d., on 100 tons of MAP1 by only one genus—Bacillus spp. — obtained by heating the sample at 80° C. for 15 minutes. Under the same conditions fewer effective results in Fe.sub.2O.sub.3 removal from 0.13 wt % to 0.10 wt % (23% removal), and the brown colour was removed only partially.

[0070] Comparison of example 1 with the prior art I. Styriakova et al., “Dissolution of iron from quartz sands by basin bioleaching under static in-situ conditions”, HYDROMETALLURGY, ELSEVIER SCIENTIFIC PUBLISHING CY. AMSTERDAM, NL., vol. 104, no. 3-4, 1 Oct. 2010, pp. 443-447 (2010):

[0071] In the process of the prior art, 49.5% of Fe.sub.2O.sub.3 was removed in the bioleaching process only with the Bacillus spp. In very contrast to this, example 1 shows a removal of 53% Fe.sub.2O.sub.3 with the mixture autochthonous and allochthonous heterotrophic bacteria and, thus, a superior efficiency.

EXAMPLE 3

[0072] The experiment of the effectiveness of autochthonous, allochthonous and a mixture of symbiosis of autochthonous and allochthonous bacteria was verified on three types of sand (Q1, Q2, Q3) from three localities (S̆ajdikové Humence, Slovakia; Moravc̆e, Slovenia; and Petrijevc̆i, Croatia) with iron content and the presence of autochthonous bacteria.

[0073] In the first case of testing autochthonous bacteria in parallel samples without the addition of active allochthonous bacteria and only by stimulating iron removal by adding medium during 1 month bioleaching, Fe reduction was achieved at Q1—20%, at Q2—28%, at Q3—27%.

[0074] In the second case, autochthonous bacteria was suppressed by sterilizing the sample and water to prepare the medium in parallel tests by: a) 5% H.sub.2O.sub.2 b) 3% NaClO and c) 30 minutes heating to 80° C. The 0.01% allochthonous mixture heterotrophic bacteria was inoculated in the concentration 10.sup.8 CFJ/ml. Fe removal increased after 1 month of bioleaching in various cases of tests a) Q1—32%, Q2—45%, Q3—42%, b) Q1—28%, Q2—39%, Q3—32%, c) Q1—36%, Q2—49%, Q3—46%.

[0075] In the third case of bioleaching without sterilization of sample and water for media preparation, the stimulating effect of using media for autochthonous and inoculated active allochthonous bacteria in parallel samples was achieved with the highest Fe removal efficiency Q1—53%, Q2—54%, Q3—52%.

Comparative Examples with Regard to the Prior Art

[0076] To compare the present invention with U.S. Pat. No. 6,395,061, a test on laterite ore with mixed autochthonous and allochthonous bacteria was carried out, wherein 45% extraction of Ni was achieved after 1 month bioleaching. In addition, we compared the bioleaching of iron oxides by Aspergillus niger which resulted in removal of 47.7% of the total iron in the quartz sand. The iron content decreased from 0.315% Fe.sub.2O.sub.3 to 0.164% Fe.sub.2O.sub.3 in the bioleached quartz sand by this fungi. In very contrast to this, 53% efficiency was achieved by the process according to the present invention.

[0077] In prior art reference RU 2 603 934 C1 no data on efficiency is provided. To compare the present invention with this prior art, an experiment on silica sand from S̆ajdikové Humence (Slovakia) with mixed culture of Bacillus spp. and Saccharomyces spp. was carried out, whereby Fe.sub.2O.sub.3 decreased by 38% (from 0,317% to 0,196%) after 1 month bioleaching (in RU 2 603 934 C1). In example 3 (third case Q1), an efficiency of 53% was achieved.

[0078] According to R. Matlakowska et al., “The culturable bacteria isolated from organic-rich black shale potentially useful in biometallurgical procedures,” JOURNAL OF APPLIED MICROBIOLOGY, vol. 107, no. 3, 30 Sep. 2009, pp. 858-866 (2009), the bioleaching is carried out by using single isolates (strains) of autochthonous bacteria (LM1-LM8). The concentrations of copper and arsenic in liquid solution during 42 days of growth of bacterial isolates on mineral medium supplemented with black shale were in range 0.7 to 2 mg/l of Cu and 0.25 to 1.25 mg/l of As after 9 days and 0.75 to 2.45 mg/l of Cu and 2.5 to 4.2 mg/l of As after 42 days. In very contrast to this, experiments according to the present invention with mixture of autochthonous and allochthonous bacteria in the frame of RIS CuRE project on copper tailings from Serbia (bor mine) achieved a much higher efficiency in Cu concentration in leachate (1369 mg/l) and 1.5 mg/l of As already after 9 day of bioleaching. Moreover, high concentrations of Fe (2829 mg/I) and other elements Zn (9.6 mg/l), Sr (9.9 mg/l), Zr (5.2 mg/l), Mo (3.1 mg/l) and Hg (11.6 mg/I) were measured in the leachates after 9 days of bioleaching.