Formulation based on bacteria of the bacillus genus, method for obtaining same and use thereof as a bioremediation agent in wastewater and fat traps

Abstract

The present invention provides a formulation based on bacteria of the Bacillus genus that are capable of generating active sludges that degrade molecules present in water / soil and which can adapt to the environment to which they are exposed. The bacteria are inoculated in an organic substrate for the growth thereof, through an aerobic fermentation process in solid medium. The formulation can reduce the lipid, fat and protein content in wastewater, as well as eliminate bad odours and coliforms from same. It is therefore proposed for use as a bioremediation agent for wastewater from nejayote, blackwater and fat traps.

Description

[0001] Formulation based on bacteria of the genus Bacillus, its production process and its use as a bioremediation agent in wastewater and grease traps

[0002] DESCRIPTION

[0003] OBJECT OF THE INVENTION

[0004] The object of the present invention is to provide a bacterial formulation containing six different strains of the genus Bacillus, which are capable of generating activated sludge that degrades molecules present in water / soil and are able to adapt to the environment to which they are exposed. The bacteria are inoculated onto an organic substrate for growth through an aerobic fermentation process in a solid medium. The formulation is capable of reducing the lipid, fat, and protein content in wastewater, as well as eliminating odors and coliform bacteria. Therefore, its use is proposed as a bioremediation agent, particularly for wastewater from the nej ayote (a type of squash), sewage, and grease traps.

[0005] BACKGROUND

[0006] The goal of wastewater treatment is to separate the organic content from wastewater in order to minimize the amount of waste and pollutants present. Nejayote, a byproduct of nixtamalization, produces wastewater with an alkaline pH (pH > 10) and high concentrations of both dissolved and suspended organic matter, making it one of the most polluted effluents discharged into bodies of water. This causes increased turbidity and decreased dissolved oxygen, negatively impacting the survival of various aquatic organisms.

[0007] In hotels, restaurants, and hospitals, grease production is often high, which is why grease traps are commonly used as wastewater pretreatment. Thanks to modern technological advancements, effluent quality and pretreatment performance can be improved by using grease traps containing enzymes such as lipases, proteases, and amylase. The purpose of these enzymes is to break down and shorten the chains of oils and fats so they can be consumed by non-pathogenic bacteria and removed from the waste stream.

[0008] Among the technologies currently used that employ a microbial formulation are Micro-Blaze® FOG, ACF-32, ECOBAC EBG, ECOBACBEOK, ECOBAC CLA, Bio-Green WT, Bio-Green HC, Bio-Green GT, Maintain® L, Munox 1, Munox 5, Bacilox XL OE, Bioalkim CO, ROETECH 106 PS, Bioseptic Master, ACBiol, BIODEGRADER, and BIODEGREASER. Those that utilize a microbial formulation supplemented with enzymes include Bioalkim TGD and MicroCat®. Finally, the one that uses a formulation using only micronutrients is Biologic® GTT (AMEREX - Bioaugmentation. (n.d.). http: / / www.labamerex.com / novedad019.htm; Micro Blaze. (2022b, January 1). Micro Blaze, https: / / www.micro-blaze.com / ; Industrial-commercial-water-filtration-products-and-services-company-puerto-rico-and-the-caribbean. (n.d.). https: / / watertechgroup.com. https: / / www.watertechgroup.com / es / ).

[0009] Products using a microbial formulation share the common characteristic of employing a mixture of wetting agents, nutrients, and various strains of safe, non-pathogenic Bacillus bacteria or non-pathogenic Pseudomonas strains (Munox 1, Munox 5, and ROETECH 106 PS). The purpose of these microorganisms is to work alongside the existing microbial community to enhance its ability to break down carbohydrates, cellulose, proteins, and fats. As a result, the microorganisms rapidly produce a wide range of enzymes that liquefy the target waste into digestible pieces for the microbes to consume, leading to a more efficient and reliable wastewater treatment process (Bacteria for Wastewater and Waste Treatment, (n.d.). Manufacturing Microbes for Crops, Livestock Waste, Aquaculture and Wastewater, https: / / www.biogp.com / tratamiento-de-aguas-residuos8203.html; Welcome to StepCleaner. (n.d.). https: / / www.stepcleaner.com / productos.php; Química Ecológica de México SA de CV (February 16, 2024). Products - Ecological Chemistry. Química Ecológica. https: / / quimicaecologica.com / productos / ; Wastewater Treatment - Products | American Biosystems. (August 7, 2019). American Biosystems. https: / / www.americanbiosystems.com / productos / tratamiento-de-aguas-residuales / ?lang=es).

[0010] Products that use a microbial formulation with added enzymes are microbial formulas applied to initiate, restore, or improve biological degradation processes. These formulas are typically composed of microorganisms (bacteria and fungi), enzymes, wetting agents, nutrients, adsorbents, and buffers (Municipal Wastewater - QM Environmental Services. (2024, March 1). QM Environmental Services, https: / / qmes.eu / producto / aguas-residuals-municipal / ?lang=es; Bioalkim Biotechnology - Bacteria for Water and Fat Treatment, (n.d.). BIOALKIM. https: / / www.bioalkim.net / ).

[0011] Finally, Biologic® GTT, which uses a formulation using only micronutrients, contains no bacteria, enzymes, or chemical additives. It is a concentrated liquid for treating grease traps and drain pipes made with micronutrients, which accelerate the digestion of organic solids, fats, oils, and tallow, eliminate odors, and improve the operation and effluent quality of grease traps (Biologics Micronutrients, bacteria for bad odors, (n.d.). It's Over 9000! https: / / biologics.com.mx / es / micronutrientes).

[0012] Regarding the prior art of the present invention, we find document number CA2313110A1, which relates to a microbiological process based on the use of novel, non-genetically modified, naturally occurring, non-pathogenic rhizobia strains that degrade or transform contaminants, suitable for the bioremediation or biorestoration of soils or water contaminated with compounds as simple as nitrate fertilizer and as complex as substituted aromatic hydrocarbons. The products obtained through the described process can also be used as biofertilizers or bioherbicides to improve the quality of agricultural and recreational sites.

[0013] There are also other documents, such as number VS6248234B1, which comprises a bioremediation device including a water-soluble material that can be maintained in a suspension network. The material preferably contains surfactants with hydrophilic and / or hydrophobic tails that can readily form emulsions with the wastewater material present through critical micell formation. These surfactants preferably include, among others, non-ionic surfactants that carry no charge. The hydrophilic portion preferably contains numerous polar ether linkages derived from the polymerization of ethylene oxide and / or propylene oxide with the hydrophobic component. The preferred material comprises biodegradable alkanolamine surfactants that remove fat and organic matter from wastewater by biological means through the induction of bacteria and enzymes as the material dissolves in the presence of wastewater.Variations in the size, shape, and quantities of bacteria in the soluble solid affect the nature of the degradation in relation to the flow, type of waste, and retention time to decrease the wastewater bioremediation time.

[0014] Patent number US6325934B1 describes a system that incorporates bacteria and digestive enzymes from wastewater into a slow-release material and delivers it to the waste site to digest solid waste. The slow-release material is made heavy so the enzymes and bacteria are delivered to the sludge at the bottom of a wastewater digestion chamber and is made fat-soluble so the enzymes and bacteria are delivered to the fat in the waste system for digestion. This selective delivery system prevents the enzymes and bacteria from being diluted in greywater, which would make them less effective and cause them to be discharged from the sewer system undigested.

[0015] Document number W02005108309A1 describes a biological treatment that can be used for bacterial bioremediation in aquatic ecosystems and urban and industrial wastewater to ensure adequate digestion of organic matter and nutrients. The treatment comprises a method for preparing and applying a bacterial inoculum to water from the aquatic environment to be treated. The inventive treatment comprises the following steps: taking water from the environment to be treated using an opaque, non-metallic reactor vessel and adding a protein hydrolysate; recirculating the water for 10 minutes; subsequently adding trace elements of iron, cobalt, zinc, nickel, and molybdenum; and finally, adding a lyophilized biological product of aerobic and facultative anaerobic bacteria at a concentration of 7 x 10⁻⁶ 9CFU / gram, along with its enzymes, and recirculate the mixture for at least 30 minutes, so that it is ready to be applied to the substrate to be treated and contains the bacterial inoculum prepared in it.

[0016] Invention number W02005042406A2 is a bioremediation product comprising a biodegradable carrier and a tablet or powder consisting essentially of microorganisms capable of digesting hydrocarbons, an inert material and, optionally, traces of oil in a sufficient quantity to maintain the microorganisms in a dormant state, said microorganisms being located entirely within said biodegradable carrier, and wherein the biodegradable carrier is in direct contact with the microorganisms.

[0017] The invention described in document number US7166221B1 is a system and method for the bioremediation of hydrocarbon and organic pollution in fresh and salt water. Microorganisms that digest the organic and hydrocarbon pollution are placed on a floating support where the microorganisms are exposed to and digest the pollution. The floating element can be a block of polymer foam. The microbes can be supported on powder, such as clay minerals, and the powder can be formed into granules held in grooves in the foam.

[0018] Document number MX351319 uses a mixture for the bioremediation of domestic wastewater comprising a granule with high bacteriological and enzymatic content which in turn comprises i) a biodegradable and porous organic substrate, ii) saprophytic bacteria of the genus Bacillus, preferably in a vegetative state and iii) lytic enzymes; and a substrate with a high starch content as the only source of nutrients for the bacteria it supports.

[0019] WO2012137220A2 provides a bioaugmentation composition for improving the hydrocarbon degradation efficiency of a wastewater treatment plant for the degradation of hydrocarbons in wastewater generated in the hydrocarbon processing industry, and a method for such degradation. The composition comprises a synergistic combination of selective microorganisms to form a consortium that enables the effective degradation of hydrocarbons present in the wastewater and their conversion into harmless and environmentally friendly substances. The invention also provides these microorganisms and their isolates.

[0020] Document number WO2014135735 describes a biological process for reducing the hydrocarbon content in an aqueous medium comprising a hydrocarbon mixture containing at least one aliphatic alkane; at least one polycyclic aromatic hydrocarbon (PAH); and at least one aromatic hydrocarbon selected from the group consisting of benzene, toluene, ethylbenzene, o-xylene, m-xylene, p-xylene, and combinations thereof. The process involves contacting the aqueous medium contaminated with the hydrocarbon mixture with a bacterial mixture comprising at least one bacterium that degrades an aliphatic alkane; at least one bacterium that degrades a PAH; and at least one bacterium that degrades the aromatic hydrocarbon. The process may include a chemical or photochemical treatment of the contaminated aqueous medium prior to the biological treatment. Bacteria capable of degrading the hydrocarbons are also described.

[0021] Document number WO2013022332 presents for protection the process of preparing a bioremediation mixture containing nixtamal flour and a composition based on bacteria of the genus Bacillus. This mixture is used for bioremediation and bioaugmentation purposes, which involve the degradation and decontamination of compounds; it also eliminates bad odors and fecal coliforms, and reduces the content of lipids, starch, and proteins present in contaminated water or solid surfaces exposed to air and moisture, as well as for the degradation of cellulose in soils and as a fertilizer additive.

[0022] The invention MX2017013555 has as its final product the generation of specialized anaerobic granules capable of removing contaminants and pathogenic biological agents from wastewater requiring treatment. These specialized anaerobic granules have the ability to function optimally in short periods of time (3-6 days) compared to what is reported in the literature (up to 30 days), due to the generation and specialization methodology used to obtain them.

[0023] Invention JP2017534245 relates to a composition comprising a bacterial spore and a germinative compound, wherein said components are maintained in an inactive form. This invention also relates to the use of the compositions in wastewater treatment, environmental remediation, oil recovery, aquaculture systems, and direct-feed microbes.

[0024] Document number US20170320763 comprises a soil and groundwater remediation composition containing halogenated compounds. The remediation composition includes an elemental iron-based composition, which may include activated carbon capable of adsorbing the halogenated compounds through numerous pores impregnated with elemental iron. It also includes a mixture of one to many organisms capable of degrading the halogenated compounds. The remediation composition includes an organic compound or polymeric substance and a second bioremediation material, which is a mixture of one to many organisms capable of degrading the organic compound or polymeric substance over time (e.g., 20 to 365 days or more to provide a release time) into smaller molecules or compounds used by the organisms in the first bioremediation material as they degrade the halogenated compounds.The organic compound can be a complex carbohydrate such as food-grade starch, chitin, or another complex carbohydrate such as one with low water solubility.

[0025] The invention RU2617953C1 is a formulation for treating soil and water from petroleum and petroleum products. It contains oil, biomass, hydrocarbons, the bacterial strain Bacillus subtilis SNBS-1, deposited in the Russian National Collection of Industrial Microorganisms under accession number VKPM B-12239; an astringent component used as an organomineral substrate enhancer; and carboxymethylcellulose, a mixture composed of sawdust prepared with clinoptilolite and humus components in a predetermined ratio. The effect of the invention is to improve the efficiency of purifying soil and water from crude oil and petroleum products.

[0026] Invention number US2017008814A1 is a biocatalytic composition for use in agricultural fields, in the animal husbandry area, and in environmental restoration areas, to transform substrates with which it reacts into non-polluting substances; this composition comprises a component with coenzymatic activity that includes vitamin A, vitamin D3, vitamin E, propyl gallate, crude fats, crude proteins; a component with enzymatic activity that includes Bacillus licheniformis, Bacillus subtilis, Bacillus thuringiensis, Bacillus spp., Aspergillus oryz.ae, Aspergillus niger, Lactobacillus bifidus, Lactobacillus acidophilus, amylase, protease, lipase, cellulase, gumase; and a component that includes pH-regulating substances, comprising humic acids, fulvic acids, choline acids, apocrenic acids, vitamin A, vitamin D3, vitamin PP, Arthrospira maxima, among others.

[0027] Document number RU2691317C1 falls within the field of biotechnology, as this invention increases the efficiency of purifying water containing fat. This invention comprises the use of a consortium based on strains of the bacteria Bacillus sp. VKM B-3288D and Pseudomonas VKM B-3287D, taken in a weight ratio of 1:1, with a final concentration of colony-forming units of bacteria in concentrated suspension of not less than 1 x 10 CFU / g.

[0028] Document number US2020102236A1 is a process for the treatment of hydrocarbon refinery wastewater that produces a low level of biosludge, including the process the utilization of microbial consortia comprising at least one species of Pseudomonas and at least one species of Bacillus in a ratio of 10:1 to 1:10. The Pseudomonas species and the Bacillus species have constitutive expression of at least one hydrocarbon-degrading gene. Pseudomonas species are selected from the group consisting of Pseudomonas stutzeri (MTCC 25027), Pseudomonas aeruginosa (MTCC 5389), Pseudomonas aeruginosa strain IOC DHT (MTCC, 5385), Pseudomonas putida IOCR1 (MTCC 5387), Pseudomonas putida IOC5al (MTCC 5388) and a mutant of the same. The Bacillus species are selected from the group consisting of Bacillus subtilis (MTCC 25026), Bacillus subtilis (MTCC 5386), Bacillus thermoleovorans (MTCC 25023), Bacillus stearothermophilus (MTCC 25030), Lysinibacillus sp.(MTCC 25029), Lysinibacillus sp. (MTCC 5666) and a mutant thereof. The microbial consortia are used at a concentration of at least 102 CFU / ml.

[0029] Invention US2020369849A1 relates to the treatment of waste with one or more microorganisms for the purpose of, among other things, degrading waste, bioremediating waste, improving waste stabilization, reducing contaminants in waste as well as its odor, reducing organic compounds in waste, and combinations thereof. More particularly, the invention relates to isolated strains of Bacillus and strains having all the identifying characteristics of these strains, and combinations thereof, for uses comprising the uses mentioned above.

[0030] The invention CN112251368A belongs to the field of environmental biotechnology and provides an immobilized microbial cell for degrading fats in highly concentrated kitchen wastewater, along with a method for its preparation and application. The microbial cell is Kluyvera cryocrescens LYC50-la. During immobilization, the microbial cell is immobilized using a sodium alginate / polyvinyl alcohol / activated carbon embedding method. The immobilized cell can be stored for a long time, reused 6 to 7 times, and can degrade peanut, used cooking, soybean, rapeseed, sesame, linseed, mixed, and sunflower seed oils. The immobilized cell has the best degradation effect on mixed oils, with a degradation rate of 95%. The immobilized microbial cell has a fat degradation rate of over 90% in kitchen wastewater.

[0031] The technology described in patent number US10981818B2 relates to an apparatus, methods, and applications for cultivating microorganisms in situ to treat contaminated environments. The apparatus is designed to operate under a wide range of environmental conditions, including extreme cold, extreme heat, and direct sunlight. These environments typically reduce the lifespan of the organisms in the storage chamber that feeds the heater where they are cultivated. These environments can also reduce the growth rate of the organisms in the fermenter, leading to decreased cell production. Often, the optimal application site for the organisms is outdoors and too far from structures with adequate protection against the sun's ultraviolet radiation or excessive cold or heat.

[0032] Document number ES2933285A1 is an application for a patent of invention comprising a purification procedure to be applied in the facilities where the discharges themselves are generated, which uses a biological treatment based on adding a mixture of bacteria, with which to digest and reduce the organic matter and nutrients, which are found in the untreated discharges, in the water or accumulated in the river sediments.

[0033] BRIEF DESCRIPTION OF THE FIGURES

[0034] Figure 1. Shows a flow diagram of the process for obtaining the formulation that is the subject of the present invention.

[0035] Figure 2 shows a graph of the consortium's capacity to reduce BOD and nitrogenous pollutants in an industrial wastewater treatment plant over a 12-week period. The results are compared with those of the clarifier in the treatment plant that does not use the consortium. The treated water data shows up to a 96% improvement in pollution reduction.

[0036] Figure 3. Shows a photograph of the appearance of the water treated with the invention, in a wastewater treatment plant in Tecomán, showing the clarity obtained in the treatment oxidation lagoon of this municipality of Colima.

[0037] Figure 4. Shows a photograph with a top view of a grease trap prior to the application of the formulation that is the subject of the present invention.

[0038] Figure 5. Shows a photograph with a top view of the trap, shown in Figure 4, three months after the weekly application of the invention, where 100 grams of the formulation were applied per week.

[0039] Figure 6. Shows a photograph of a 1000 m³ oxidation pond 3 , where the invention was applied. DETAILED DESCRIPTION OF THE INVENTION

[0040] The present invention provides a formulation based on bacteria of the genus Bacillus, its process of obtaining it, and its use as a bioremediation agent in wastewater and grease traps, in particular.

[0041] As shown in Table 1, the formulation of the present invention comprises 19 to 38% w / w of a primary mixture based on spores and thermostable enzymes derived from six species of the genus Bacillus, 31 to 69% w / w of a diluent, and 12% vegetable soil; wherein said primary mixture based on spores and thermostable enzymes derived from bacteria of the genus Bacillus comprises a wheat bran granule with IxlO 9spores / g of wheat bran, said diluent being selected from wheat flour, agricultural cellulose residues and a mixture thereof in a 5:1 ratio; and the topsoil being selected from soil, humus or a mixture thereof in a 0.2:1 ratio i.e. one tenth to one of the formulation; wherein the species of the genus Bacillus comprise 5 to 20% of Bacillus subtilis, 5 to 20% of Bacillus megaterium, 5 to 20% of Bacillus polymyxa, 5 to 20% of Bacillus cercus, Bacillus licheniformis, 5 to 20% of Bacillus sp, preferably Bacillus cereus, as shown in Table 2, and the thermostable enzymes comprise proteases, lipases, amylases, cellulases, of different characteristics.

[0042] Table 1. Percentage included of each element of the formulation of the present invention

[0043]

[0044] Table 2. Composition of the primary mixture present in the invention.

[0045] The formulation of the present invention utilizes six Bacillus strains with different metabolic strategies and capabilities. These strains are exposed to highly contaminated environments, and depending on the source of contamination, only those strains possessing the metabolic capabilities for the biological niche to which they are exposed grow and degrade the organic pollutants. This microbiological consortium has the capacity to generate activated sludge that continues to degrade the molecules present in the water / soil and adapts to the environment to which it is exposed. The formulation of the present invention is capable of reducing the lipid, fat, and protein content in wastewater, as well as eliminating unpleasant odors and coliform bacteria. The bacteria are inoculated onto an organic substrate for growth, preferably wheat bran, through an aerobic fermentation process in a solid medium.

[0046] The process for obtaining the formulation based on bacteria of the genus Bacillus, the subject of the present invention, comprises the steps of: a) Sterilizing wheat bran in an autoclave for 15 min at 120°C. b) Aerobically fermenting the wheat bran sterilized in step a) in a solid medium and inoculating it with spores of six species of the genus Bacillus, inoculating 1X10 3 1X10 5spores / g of bran, in a tray reactor, in a layer with a height less than or equal to 1.5 cm, at a temperature between 37 to 38° C for 7 to 12 hours to obtain a premix, wherein the six species of the genus Bacillus comprise 5 to 20% of Bacillus subtilis, 5 to 20% of Bacillus megaterium, 5 to 20% of Bacillus polymyxa, 5 to 20% of Bacillus cereus, Bacillus licheniformis, 5 to 20% of Bacillus sp. preferably Bacillus cereus.c) Dry the primary premix obtained in step b) at 90°C for 24 hours to obtain a primary mixture based on spores and heat-stable enzymes derived from six species of the genus Bacillus; and d) Add topsoil and a pest-free organic diluent to obtain a formulation based on bacteria of the genus Bacillus used as a bioremediation agent, with a proportion of 19 to 38% w / w of a primary mixture based on spores and heat-stable enzymes derived from six species of the genus Bacillus, 31 to 69% w / w of a pest-free, heat-dryable organic diluent, and 12% topsoil, wherein the pest-free, heat-dryable organic diluent is selected from wheat flour, agricultural cellulose residues, or a mixture thereof in a 1:1 ratio, and the topsoil is selected from soil, humus, or a mixture thereof. a 1:1 ratio.

[0047] EXAMPLE 1. PROCESS FOR OBTAINING THE FORMULATION BASED ON A MICROBIAL CONSORTIUM

[0048] Solid-state fermentation (SSF) processes have a wide range of applications in the growth of aerobic organisms, bioremediation, and degradation of organic solid waste, as well as in the production of products such as enzymes, antibiotics, or pigments. They involve the growth of microorganisms on moist solid particles and, in contrast to liquid fermentation systems, use a minimal amount of water (Mitchell, D., Krieger, N., & Bevorick, M. (2006). Solid-State Fermentation Bioreactors. Springer-Verlag. Germany).

[0049] However, it also has disadvantages that must be taken into account, such as operation at temperatures higher than the optimum due to metabolic heat, temperature gradients due to poor mixing, inadequate mixing to minimize the shear stress to which the microorganism is subjected, and substrate dehydration caused by the effect of increased temperature and airflow. Therefore, this type of system is used when the product is in solid form (as in the present invention), when production is higher than in liquid systems, when socioeconomic conditions allow it, or when a solid residue is used to minimize environmental impacts. Thanks to previous studies on wheat bran, it is possible to conclude that solid-state fermentation is suitable for the purposes of the present invention (Aguirre, R. (2008).Confidential report of the experiences at UAM-I that supported the patent of Viniegra-González, et al. 2011; Viniegra-González, G., Favela-Torres, E., Aguirre-Gamboa, R., Aréchiga-Carvajal, E. & Adame-Rodríguez, J. (2011). Microorganism-based mixture for bioremediation, manufacturing process and use. IMPI Patent Registration).

[0050] To obtain 1 kg of the formulation, 1 kg of wheat bran is sterilized and then placed in trays with a height less than or equal to 1.5 cm. The wheat bran is fermented with strains of the genus Bacillus at a temperature between 37° and 38° C in an oven.

[0051] It is dried at 90°C for 24 hours to obtain thermostable spores and enzymes, resulting in a primary mixture based on spores and thermostable enzymes derived from six species of the genus Bacillus. Humus and an organic diluent are added; this organic diluent is selected from flours and cellulose residues of agricultural origin, which allows the spores to enter a vegetative state during the first 12 hours before the product is applied.

[0052] It must be verified that the diluent does not attract pests and can be heat-dried to extend shelf life for obtaining the formulation with the active ingredient. The fermented product is produced in solid phase. After fermentation, a growth curve for the microorganisms is generated. Figure 1 shows a flow diagram of the process for obtaining the formulation based on a microbial consortium. EXAMPLE 2. USE OF THE FORMULATION FOR BIOREMEDIATION OF WASTEWATER FROM NEJA VOTE

[0053] For the treatment of wastewater from nej ayote, the formulation is supplemented with twice the amount of the primary mixture based on spores and thermostable enzymes derived from six species of the genus Bacillus, and that amount is subtracted from the diluent, as shown in Table 3, to obtain this modality of the invention:

[0054] Table 3. Percentage included of each element of the formulation when used for the bioremediation of wastewater from Nejayote.

[0055] For the treatment of nejayote water, a concentration of 1g of component per liter of nejayote water was used, and 72 hours after application, another gram per liter was added. The following results were obtained, shown in Table 4, where the percentage of degradation achieved with the present invention is 36% in 72 hours.

[0056] Table 4. Results of the nejayote water treatment. S / T: Untreated EXAMPLE 3. USE OF THE FORMULATION IN GREASE TRAPS

[0057] Using 100 grams of formulation each week in 40m grease traps 3For three months, the grease trap remains clean without the need for manual cleaning. Figure 4 shows the trap before the application of the invention, and Figure 5 shows the same trap clean after weekly application of the invention for three months.

[0058] EXAMPLE 4. USE OF THE FORMULATION IN WASTEWATER

[0059] The invention was used in a wastewater treatment plant in Tacomán, Colima. The results are detailed in Table 4, where a considerable decrease in the influent and effluent values ​​for Total Suspended Solids (TSS), Volatile Suspended Solids (VSS), and Chemical Oxygen Demand (COD) is observed after the use of the present invention. The appearance of the water is shown in Figure 3.

[0060] Table 4. Results of the use of the present invention in a wastewater treatment plant in Tacoman, Colima

[0061] The formula, which is the subject of the present invention, was used for three months in a 1000 m³ oxidation pond. 3 , shown in figure 6, (at a rate of 1 kg per m 3 Add the formulation to the water each week to replace the water lost through carryover, eliminating odor and improving the overall condition of the water.

[0062] EJEMPLO 5. DEGRADACIÓN DE NITRÓGENO AMONIACAL

[0063] The formulation degrades ammonia nitrogen to 25 ppm. This test was followed for 12 weeks in a test bioreactor with a minimum retention time of 24 hours, of 100m 3 Using a shock dose of 1 kg and daily replenishment of 100 g of the formulated solution. Nitrogen concentrations and BOD were determined at least three days a week; the results of this application of the invention are shown in Table 5.

[0064]

[0065] Table 5. Results of the application of the invention to degrade ammoniacal nitrogen

Claims

CLAIMS Having sufficiently described my invention, I consider as a novelty, and therefore claim as my exclusive property, the contents of the following clauses:

1. A formulation based on bacteria of the genus Bacillus characterized in that it comprises 19 to 38% w / w of a primary mixture based on thermostable spores and enzymes derived from six species of the genus Bacillus, 31 to 69% w / w of a diluent of organic origin that is pest-free and capable of being heat-dried, and 12% topsoil; wherein said primary mixture based on spores and thermostable enzymes derived from six species of the genus Bacillus comprises a wheat bran granule with IxlO 9spores / g of wheat bran, the diluent of organic origin is chosen from wheat flour, cellulose residues of agricultural origin or a mixture thereof, in a 1:1 ratio, and the topsoil is chosen from soil, humus or a mixture thereof in a 1:1 ratio; wherein the species of the genus Bacillus comprise 5 to 20% of Bacillus subtilis, 5 to 20% of Bacillus megaterium, 5 to 20% of Bacillus polymyxa, 5 to 20% of Bacillus cercus, Bacillus licheniformis, 5 to 20% of Bacillus sp, preferably Bacillus cereus, and the thermostable enzymes comprise proteases, lipases, amylases, cellulases, of different characteristics.

2. The formulation based on bacteria of the genus Bacillus, according to claim 1, for use as a bioremediation agent.

3. The formulation based on bacteria of the genus Bacillus, according to claim 2, wherein the bioremediation agent is for wastewater from nejayote.

4. The formulation based on bacteria of the genus Bacillus, according to claim 2, wherein the bioremediation agent is for sewage.

5. The formulation based on bacteria of the genus Bacillus, according to claim 2, wherein the bioremediation agent is for grease traps.

6. The formulation based on bacteria of the genus Bacillus, according to claim 2, wherein the bioremediation agent degrades ammonia nitrogen to 25 ppm.

7. The formulation based on bacteria of the genus Bacillus, according to claim 3, wherein when the bioremediation agent is for wastewater from nej ayote the formulation comprises 38% w / w of the primary mixture based on thermostable spores and enzymes derived from six species of the genus Bacillus, 50% w / w of the diluent and 12% w / w of vegetable soil.

8. The formulation based on bacteria of the genus Bacillus, according to claim 3, wherein when the bioremediation agent is applied to wastewater from nej ayote, a 36% degradation occurs in 72 hours.

9. A process for obtaining the formulation based on bacteria of the genus Bacillus, according to claim 1, characterized in that it comprises the steps of a) Sterilizing wheat bran in an autoclave for 15 min at 120 °C. b) Aerobically fermenting the wheat bran sterilized in step a) in a solid medium and inoculating it with six species of the genus Bacillus, 1X10 3 1X10 5spores / g of bran, in a tray reactor, in a layer with a height less than or equal to 1.5 cm, at a temperature between 37 to 38° C for 7 to 12 hours to obtain a premix, wherein the six species of the genus Bacillus comprise 5 to 20% of Bacillus subtilis, 5 to 20% of Bacillus megaterium, 5 to 20% of Bacillus polymyxa, 5 to 20% of Bacillus cereus, Bacillus licheniformis, 5 to 20% of Bacillus sp. preferably Bacillus cereus c) Dry the primary premix obtained in step b) at 90°C for 24 hours to obtain a primary mixture based on spores and thermostable enzymes derived from six species of the genus Bacillus', d) Add topsoil and a pest-free organic diluent to obtain a formulation based on Bacillus bacteria used as a bioremediation agent, with a proportion of 19 to 38% w / w of a primary mixture based on thermostable spores and enzymes derived from six species of the genus Bacillus, 31 to 69% w / w of a pest-free organic diluent capable of being heat-dried, and 12% topsoil.

10. The process for obtaining the formulation based on bacteria of the genus Bacillus, according to claim 9, characterized in that the diluent of organic origin, pest-free and capable of being heat-dried, is selected from wheat flour, agricultural cellulose residues, or a mixture thereof in a 1:1 ratio.

11. The process of obtaining the formulation based on bacteria of the genus Bacillus, in accordance with claim 9, characterized in that the topsoil is chosen from soil, humus or a mixture thereof in a 1:1 ratio.

12. The process of obtaining the formulation based on bacteria of the genus Bacillus, in accordance with claim 9, characterized in that when the formulation based on a microbial consortium is used for the treatment of wastewater from nej ayote, twice the amount of active ingredient is added and less of the diluent is subtracted.

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