MICROORGANISM MIXTURE, COMPOSITION, PROCESS FOR THE PREPARATION THEREOF, METHOD AND USE IN TREATING SOLID ORGANIC WASTE

Abstract

The present invention relates to a microorganism mixture comprising one or more microorganisms selected from the Bacillus, Paenibacillus, lactobacillus, Pseudomonas, Trichoderma, aspergillus, and Saccharomyces genera, and which is useful in treating of solid organic waste. The present invention also relates to a composition comprising said microorganism mixture, a process for preparing said composition, a method for treating solid organic waste, and the use of the microorganism mixture or the composition of the invention in treating solid organic waste.

Claims

1. A microorganism mixture comprising two or more microorganisms selected from the group consisting of Bacillus, Paenibacillus, lactobacillus, Pseudomonas, Trichoderma, aspergillus, and Saccharomyces genera.

2. The microorganism mixture according to claim 1, comprising two or more microorganisms selected from the group consisting of Bacillus subtilis, Bacillus licheniformis, Paenibacillus polymyxa, Lactobacillus acidophilus, Pseudomonas putida, Bacillus pumilus, Trichoderma harzianum, Aspergillus brasiliensis, Bacillus megaterium, Lactobacillus plantarum, and Saccharomyces cerevisiae species.

3. The microorganism mixture according to claim 1, comprising microorganisms of Bacillus subtilis, Bacillus licheniformis, Paenibacillus polymyxa, Lactobacillus acidophilus, Pseudomonas putida, Bacillus pumilus, and Trichoderma harzianum species.

4. The microorganism mixture according to claim 1, comprising microorganisms of Bacillus subtilis, Bacillus licheniformis, Paenibacillus polymyxa, Lactobacillus acidophilus, Pseudomonas putida, Bacillus pumilus, and Aspergillus brasiliensis species.

5. The microorganism mixture according to claim 1, characterized in that it is for use in treating solid organic waste.

6. A composition comprising the microorganism mixture.

7. The composition according to claim 6, further comprising one or more ingredients selected from the group consisting of a drier, a preservative, and a dispersant.

8. The composition according to claim 7, wherein the drier is selected from the group consisting of kaolin, silicon dioxide, diatomaceous earth, bentonite, agalmatolite, calcium carbonate, magnesium carbonate, calcium silicate, autoclaved bone meal, and talc.

9. The composition according to claim 7, wherein the preservative is selected from the group consisting of NaCl, sodium lactate, potassium lactate, citric acid, seaweed extract, and silicon dioxide.

10. The composition according to claim 7, wherein the dispersant is selected from the group consisting of a cereal bran, carboxymethyl cellulose (CMC), hydroxymethyl cellulose (HMC), bentonite, aluminum silicate, and magnesium silicate.

11. The composition according to claim 6, comprising: 5×10.sup.7 CFU/g of Bacillus subtilis; 5×10.sup.7 CFU/g of Bacillus licheniformis; 4×10.sup.7 CFU/g of Paenibacillus polymyxa; 3×10.sup.7 CFU/g of Lactobacillus acidophilus; 3×10.sup.7 CFU/g of Pseudomonas putida; 3×10.sup.7 CFU/g of Bacillus pumilus; and 3×10.sup.7 CFU/g of Trichoderma harzianum or Aspergillus brasiliensis.

12. The composition according to claim 6, intended for use in treating solid organic waste.

13. A process for preparing the composition according to claim 6, comprising the following steps (c) to (g): (c) adding each microorganism to a culture medium (broth); (d) preparing the inoculum with the product obtained from step (c); (e) fermenting the inoculum obtained from step (d); (f) drying the fermented material obtained from step (e), thus obtaining each dried microorganism active ingredient; and (g) mixing the dry active ingredients obtained from step (f) and optionally a preservative and/or a dispersant until a homogeneous composition is obtained.

14. The process according to claim 13, wherein: i) in step (d), inoculum preparation is performed in an incubator chamber with orbital agitation (shaker) according to the following parameters: rotation from 70 to 90 rpm, preferably from 78 to 82 rpm, more preferably 80 rpm; incubation temperature from 34 to 38° C., preferably 35.5 to 36.5° C., more preferably 36° C.; fermentation time from 44 to 54 hours, preferably from 48 to 50 hours, more preferably 49 hours; and pH from 6.5 to 7.9, preferably from 6.9 to 7.5, more preferably 7.2; ii) in step (e), fermentation is performed by transferring each inoculum to an industrial fermentation tank filled with mash and by proceeding with batch fermentation, according to the following parameters: rotation from 80 rpm to 120 rpm, preferably 95 rpm to 105 rpm, more preferably 100 rpm; incubation temperature from 34 to 38° C., preferably 35.5 to 36.5° C., more preferably 36° C.; fermentation time from 161 to 181 hours, preferably from 168 to 174 hours, more preferably 171 hours; and pH from 6.5 to 7.9, preferably from 6.9 to 7.5, more preferably 7.2; iii) in step (f), drying is performed by contact with a drier; and/or iv) in step (g), mixing is performed in a Ribbon Blender mixer.

15. The process according to claim 13, further comprising the following steps (h) to (j): (h) performing analysis of microorganism concentration in CFU/g of the composition obtained from step (g), assessing if the following parameters are met: heterotrophic bacteria: 2.0×10.sup.8 to 3.5×10.sup.8 CFU/g; viable spores: 1.0×10.sup.7 to 2.5×10.sup.7 CFU/g; and conidia count: about 1.0×10.sup.7 conidia/g; (i) performing analysis for pathogenic contaminating microorganisms, such as Escherichia coli, Pseudomonas aeruginosa, salmonella spp., and Staphylococcus aureus in each batch of finished product, wherein sampling from each batch of finished product follows the formula √{square root over (N)}+1, wherein N is the amount of finished product, in kilograms or units, per batch; and (j) packing the product obtained from step (g), preferably with packing made from virgin plastic.

16. The process according to claim 13, further comprising the following steps (a) and (b): (a) performing analysis for pathogenic contaminating microorganisms, such as Escherichia coli, Pseudomonas aeruginosa, salmonella spp., and Staphylococcus aureus in each batch of raw material (dryer, preservative, and dispersant); and (b) performing moisture analysis on each batch of raw material, wherein step (b) is preferably performed by means of oven drying, Thermogravimetric analysis (TGA) or Karl Fischer Titration.

17. (canceled)

18. A method for treating solid organic waste, comprising placing said waste in contact with the microorganism mixture according to claim 1 or the composition according to claim 6.

19. (canceled)

20. The method according to claim 18, wherein the solid organic waste is food waste.

21. The method according to claim 18, further comprising allowing accelerated digestion of the solid organic waste, wherein said accelerated digestion is an aerobic process.

22. The method according to claim 18, wherein placing said waste is performed in an equipment equipped with controlled aeration and mixing resources.

23. (canceled)

Description

DETAILED DESCRIPTION

[0019] The present invention relates to a microorganism mixture comprising two or more microorganisms selected from the Bacillus, Paenibacillus, Lactobacillus, Pseudomonas, Trichoderma, aspergillus, and Saccharomyces genera.

[0020] In one embodiment, the microorganism mixture comprises two or more microorganisms selected from the Bacillus subtilis, Bacillus licheniformis, Paenibacillus polymyxa, Lactobacillus acidophilus, Pseudomonas putida, Bacillus pumilus, Trichoderma harzianum, Aspergillus brasiliensis, Bacillus megaterium, Lactobacillus plantarum, and Saccharomyces cerevisiae species.

[0021] In one specific embodiment, the microorganism mixture comprises microorganisms from the Bacillus subtilis, Bacillus licheniformis, Paenibacillus polymyxa, Lactobacillus acidophilus, Pseudomonas putida, Bacillus pumilus, and Trichoderma harzianum species.

[0022] In another specific embodiment, the microorganism mixture comprises microorganisms from the Bacillus subtilis, Bacillus licheniformis, Paenibacillus polymyxa, Lactobacillus acidophilus, Pseudomonas putida, Bacillus pumilus, and Aspergillus brasiliensis species.

[0023] The present invention further relates to a microorganism mixture defined herein for use in treating solid organic waste.

[0024] In one embodiment, the solid organic waste is food waste.

[0025] In one embodiment, treating solid organic waste consists of accelerated waste digestion, wherein said accelerated digestion is an aerobic process, optionally with controlled aeration.

[0026] In one embodiment, treating solid organic waste is performed in equipment equipped with controlled aeration and mixing resources.

[0027] The present invention further relates to a composition comprising a microorganism mixture defined herein.

[0028] In one embodiment, the composition further comprises one or more ingredients selected from a drier, a preservative, and a dispersant.

[0029] In one embodiment, the drier is selected from kaolin, silicon dioxide, diatomaceous earth, bentonite, agalmatolite, calcium carbonate, magnesium carbonate, calcium silicate, autoclaved bone meal. and talc. In one specific embodiment, the drier is kaolin.

[0030] In one embodiment, the preservative is selected from NaCl, sodium lactate, potassium lactate, citric acid, algae extract, and silicon dioxide. In one specific embodiment, the preservative is NaCl.

[0031] In one embodiment, the dispersant is selected from a cereal bran (including wheat bran, corn bran, and rice bran), carboxymethyl cellulose (CMC), hydroxymethyl cellulose (HMC), bentonite, aluminum silicate, and magnesium silicate. In a specific embodiment, the dispersant is food-grade wheat bran.

[0032] In one embodiment, the composition comprises, in % by weight, 31.6% to 33.2%, preferably 32.4%, of dryer, 13% to 17%, preferably 15%, of preservative, and 43% to 45%, preferably 44%, of dispersant.

[0033] In one embodiment, the composition comprises:

[0034] 5×10.sup.7 CFU/g of Bacillus subtilis;

[0035] 5×10.sup.7 CFU/g of Bacillus licheniformis;

[0036] 4×10.sup.7 CFU/g of Paenibacillus polymyxa;

[0037] 3×10.sup.7 CFU/g of Lactobacillus acidophilus;

[0038] 3×10.sup.7 CFU/g of Pseudomonas putida;

[0039] 3×10.sup.7 CFU/g of Bacillus pumilus;

[0040] 3×10.sup.7 CFU/g of Trichoderma harzianum or Aspergillus brasiliensis.

[0041] The present invention further relates to a composition defined herein for use in treating solid organic waste.

[0042] In one embodiment, treating solid organic waste consists of accelerated waste digestion, wherein said accelerated digestion is an aerobic process, optionally with controlled aeration.

[0043] In one embodiment, treating solid organic waste is performed in equipment equipped with controlled aeration and mixing resources.

[0044] The present invention also relates to a process for preparing the composition defined herein, wherein the process comprises the following steps (c) to (g):

[0045] (c) adding each microorganism to a culture medium (broth);

[0046] (d) preparing the inoculum with the product obtained from step (c);

[0047] (e) fermenting the inoculum obtained from step (d);

[0048] (f) drying the fermented material obtained in step (e), thus obtaining each dried microorganism active ingredient; and

[0049] (g) mixing the dry active ingredients obtained from step (f) and optionally a preservative and/or a dispersant until a homogeneous composition is obtained.

[0050] In one embodiment, in step (c) of the process of the present invention, the culture medium comprises 0.2% to 0.4%, preferably 0.3% v/v casein, 0.4% to 0.6%, preferably 0.5% v/v yeast extract, 0.005% v/v manganese sulfate tetrahydrate, 0.005% v/v magnesium sulfate heptahydrate, 0.005% v/v calcium chloride hexahydrate, 0.005% v/v potassium phosphate dibasic, 0.3% to 0, 5% v/v agar (depending on medium's degree of gelation) and distilled water enough for 1 L and pH=7.0±0.2.

[0051] In one embodiment, in step (d) of the process of the present invention, inoculum preparation is performed in an incubator chamber with orbital agitation (shaker) according to the following parameters:

[0052] rotation from 70 to 90 rpm, preferably from 78 to 82 rpm, more preferably 80 rpm;

[0053] incubation temperature from 34 to 38° C., preferably 35.5 to 36.5° C., more preferably 36° C.;

[0054] fermentation time from 44 to 54 hours, preferably from 48 to 50 hours, more preferably 49 hours; and

[0055] pH from 6.5 to 7.9, preferably from 6.9 to 7.5, more preferably 7.2.

[0056] In one embodiment, in step (e) of the process of the present invention, fermentation is performed by transferring each inoculum to an industrial fermentation tank filled with mash and by proceeding with batch fermentation, according to the following parameters:

[0057] rotation from 80 rpm to 120 rpm, preferably 95 rpm to 105 rpm, more preferably 100 rpm;

[0058] incubation temperature from 34 to 38° C., preferably 35.5 to 36.5° C., more preferably 36° C.;

[0059] fermentation time from 161 to 181 hours, preferably from 168 to 174 hours, more preferably 171 hours; and

[0060] pH from 6.5 to 7.9, preferably from 6.9 to 7.5, more preferably 7.2.

[0061] In one embodiment, the mash comprises 0.24% to 0.32%, preferably 0.28% w/w of brown sugar, 0.1% to 0.3%, preferably 0.2% w/w of plasma, 0.1% to 0.3%, preferably 0.2% w/w of starch, 0.02% to 0.06%, preferably 0.04% w/w of fat (lard), 0.01% to 0.03%, preferably 0.02% w/w of cooking salt, 0.1% to 0.3%, preferably 0.2% w/w of soy protein, 0.01% to 0.01%, preferably 0.02% w/w of calcium chloride, 0.010% to 0.014%, preferably 0.012% w/w of zinc sulfate, 0.006% to 0.010%, preferably 0.008% of ferrous sulfate, 0.002% to 0.006%, preferably 0.004% w/w of cobalt sulfate and filtered and dechlorinated water in sufficient amount to complete the volume of the fermentation tank. In one embodiment, the fermentation tank has a nominal capacity of about 250 L.

[0062] In one embodiment, in step (f) of the process of the present invention, drying is performed by contacting a drier selected from kaolin, silicon dioxide, diatomaceous earth, bentonite, agalmatolite, calcium carbonate, magnesium carbonate, silicate of calcium, autoclaved bone meal, and talc. In one specific embodiment, the drier is kaolin. In one specific embodiment, kaolin is used in a ratio of about 21% of fermented material to about 79% of kaolin.

[0063] In one embodiment, in step (g) of the process of the present invention, mixing is performed in a Ribbon Blender mixer.

[0064] In one embodiment, in step (g) of the process of the present invention, the mixture is made with 40% to 42% of dry active ingredients obtained in step (f), 13 to 17% of preservative and 43% a 45% of dispersant, in % by weight.

[0065] In one embodiment, the composition obtained in step (g) comprises, by in % by weight, 31.6% to 33.2%, preferably 32.4% of desiccant, 13% to 17%, preferably 15% of preservative, and 43% to 45%, preferably 44% of dispersant.

[0066] In one embodiment, the dispersant is a cereal bran, preferably wheat bran, corn bran, or rice bran, more preferably food-grade wheat bran.

[0067] In one embodiment, the process according to the present invention further comprises the following steps (h) to (j):

[0068] (h) performing analysis of microorganism concentration in CFU/g of the composition obtained from step (g), assessing if the following parameters are met:

[0069] heterotrophic bacteria: 2.0×10.sup.8 to 3.5×10.sup.8 CFU/g;

[0070] viable spores: 1.0×10.sup.7 to 2.5×10.sup.7 CFU/g;

[0071] conidia count: about 1.0×10.sup.7 conidia/g;

[0072] (i) performing analysis for pathogenic contaminating microorganisms, such as Escherichia coli, Pseudomonas aeruginosa, salmonella spp., and Staphylococcus aureus in each batch of finished product, in which sampling from each batch of finished product follows the formula √{square root over (N)}+1, in which N is the amount of finished product, in kilograms or units, per lot, using a methodology described, for example, in Brazilian Pharmacopoeia, 4.sup.th edition, 1998; Brazilian Pharmacopoeia, 5.sup.th edition, 2010; United States Pharmacopeia USP 36, 2012; United States Pharmacopeia and National Formulary USP 41-NF 36, 2018; United States Pharmacopeia and National Formulary USP 41-NF 36, Method 62, 2018, Cosmetic, Toiletry, and Fragrance Association (CTFA) Microbiology Guidelines, 2007; CTFA Microbiology Guidelines, Section 19 M-2, 2007, incorporated herein by reference; and

[0073] (j) packing the product obtained from step (g), preferably with packing made from virgin plastic.

[0074] In one embodiment, the process according to the present invention further comprises the following steps (a) and (b):

[0075] (a) performing analysis for pathogenic contaminating microorganisms, such as Escherichia coli, Pseudomonas aeruginosa, salmonella spp., and Staphylococcus aureus in each batch of raw material (dryer, preservative, and dispersant), in which the sampling of each batch of raw material follows the formula √{square root over (N)}+1, in which N is the amount of raw material, in kilograms or units, per batch, employing, for example, a methodology described in step (i) above; and

[0076] (b) performing moisture analysis on each batch of raw material, verifying that moisture is between about 8% and about 10% by weight,

[0077] in which step (b) may be performed by means of oven drying, Thermogravimetry analysis (TGA), Karl Fischer Titration or any other suitable method, preferably by means of oven drying.

[0078] The present invention also relates to a composition produced by the process defined herein.

[0079] The present invention also relates to a method for treating solid organic waste which comprises contacting said waste with a microorganism mixture or a composition defined herein.

[0080] In one embodiment, the solid organic waste is food waste.

[0081] In one embodiment, treating solid organic waste consists of accelerated waste digestion, wherein said accelerated digestion is an aerobic process, optionally with controlled aeration.

[0082] In one embodiment, treating solid organic waste is performed in equipment equipped with controlled aeration and mixing resources.

[0083] The present invention also relates to the use of a microorganism mixture or a composition defined herein in treating solid organic waste.

[0084] In one embodiment, the solid organic waste is food waste.

[0085] In one embodiment, treating solid organic waste consists of accelerated waste digestion, wherein said accelerated digestion is an aerobic process, optionally with controlled aeration.

[0086] In one embodiment, treating solid organic waste is performed in equipment equipped with controlled aeration and mixing resources.

[0087] Treating solid organic waste according to the present invention provides a total reduction in the volume of solid organic waste, converting said waste into nutrient-rich gray water, which can be recycled or safely disposed of in sewage systems.

[0088] The microorganism mixture according to the present invention enhances the naturally occurring process of aerobic digestion of organic waste, causing digestion to occur in an accelerated manner.

[0089] The microorganism mixture of the invention is capable of treating most typical solid organic waste, particularly food waste, within 24 hours or less. In addition, waste treatment can be performed at the same site in which it is produced, using appropriate equipment, providing savings when compared to currently existing processes in relation to the storage, collection, transport and disposal of waste in sanitary landfills, in addition to being an environmentally friendly solution.

[0090] The present invention can be used in several places in which solid organic waste is generated, such as restaurants and bars, industrial restaurants, supermarkets, shopping centers, industries, hospitals, schools and universities, clubs and associations, residences, residential complexes, maritime vessels and rivers, or any place in which solid organic waste is generated, especially food waste.

[0091] It is worth noting that, in the currently existing processes, solid organic waste is collected, transported, and sent to landfills, which is harmful to the environment, as these processes (i) generate greenhouse gases, (ii) are anaerobic processes generating odors, (iii) are slow decomposition processes, allowing the attraction and propagation of disease vectors (insects, rodents, etc.), as well as proliferation of pathogenic microorganisms.

[0092] The microorganism mixture according to the present invention provides economic and environmental advantages, thus overcoming the inconveniences of currently existing processes through accelerated aerobic digestion.

[0093] As can be seen, the present invention overcomes the disadvantages and drawbacks of the state of the art. It is worth noting that the following examples should not be considered as limiting of the present invention, since one skilled in the art is fully capable of understanding that modifications can be made within the scope of the invention.

[0094] The entire contents of all references (patent or non-patent) cited throughout this application are hereby incorporated by reference, in particular for the teaching of the invention disclosed herein.

EXAMPLES

[0095] The examples presented herein are non-exhaustive, serving only to illustrate the invention and should not be used as limiting.

Comparative Example 1

[0096] In comparative field tests performed with products available on the market intended to degrade organic matter, such as Enzmax (Klasta Tecnologia Ambiental Ltda), Enzilimp (Millenium Tecnologia Ambiental Ltda.) and Biotrat (RZK Quimica do Brasil Ltda.), the microorganism mixture according to the present invention demonstrated a superior performance in biodegradation of food residues having diverse compositions (varying in carbohydrate, protein, lipid, and fiber contents) with a biodegradation rate such that the entire waste digestion process was performed in less than 24 hours, as well as stable biofilm forming ability and superior sporulation and recolonization ability, delivering longer lasting performance.

Example 1: Composition Preparation

[0097] Each of the following microorganisms was added to an Erlenmeyer flask filled with culture medium: Bacillus subtilis, Bacillus licheniformis, Paenibacillus polymyxa, Lactobacillus acidophilus, Pseudomonas putida, Bacillus pumilus, and Aspergillus brasiliensis, in which the culture medium consisted of 0.3% v/v of casein, 0.5% v/v of yeast extract, 0.005% v/v of manganese sulfate tetrahydrate, 0.005% v/v of sodium sulfate magnesium heptahydrate, 0.005% v/v of calcium chloride hexahydrate, 0.005% v/v of dibasic potassium phosphate, 0.4% of agar and enough distilled water for 1 L and pH=7.0.

[0098] The vials were transferred to a shaker and the preparation of the inoculum proceeded according to the following parameters:

[0099] rotation: 80 rpm;

[0100] incubation temperature: 36° C.;

[0101] fermentation time: 49 hours; and

[0102] pH: 7.2.

[0103] The final counting of microorganisms obtained in each inoculum was 2.0×10.sup.9 CFU/mL. Microorganisms were counted using microbiological sample preparation procedures, seeding techniques and serial decimal dilution counting methodology, based on the Brazilian Pharmacopoeia, 5.sup.th edition, 2010.

[0104] Each inoculum was then transferred to a 250 L nominal capacity industrial fermentation tank filled with mash, in which the mash consisted of 0.28% w/w of brown sugar, 0.2% w/w of plasma, 0.2% w/w of starch, 0.04% w/w of fat (lard), 0.2% w/w of table salt, 0.02% w/w of soy protein, 0.02% w/w of calcium chloride, 0.012% w/w of zinc sulfate, 0.008% w/w of ferrous sulfate, 0.004% of cobalt sulfate, and filtered and dechlorinated water in an amount sufficient to complete the volume of the fermentation tank. Then the fermentation proceeded in batches, according to the following parameters:

[0105] rotation: 100 rpm;

[0106] incubation temperature: 36° C.;

[0107] fermentation time: 171 hours; and

[0108] pH: 7.2.

[0109] The final counting of microorganisms obtained in the fermentation step was 3.5×10.sup.10 CFU/mL.

[0110] Each fermented material was then dried by contact with kaolin, thus obtaining each dry microorganism active ingredient.

[0111] The obtained dry active ingredients were mixed with 15% by weight of NaCl and 44% by weight of food-grade wheat bran in a Ribbon Blender mixer until a homogeneous composition was obtained, in which the percentages by weight are in relation to the total weight of the composition.

Example 2: Food Waste Treatment

[0112] The microorganism mixture obtained according to Example 1 was tested in equipment capable of controlling aeration, humidity, and temperature conditions, treating various food wastes having variable composition, containing 50% to 60% of carbohydrates, 15% 20% of proteins, 20 to 30% of lipids and 2% to 15% of fibers, using 0.1% to 0.4%, in % by weight, of the microorganism mixture.

[0113] In every case, the food waste digestion process began immediately after the equipment's environment was properly colonized and biofilms were created on walls and media by the microorganisms contained in the formulation. An efficient and accelerated food waste digestion was observed in less than 24 hours with a reduction in the volume of organic matter above 99.5%, which was converted into gray water.