CA2973961A1 - Soil inoculant and method of producing same - Google Patents
Soil inoculant and method of producing same Download PDFInfo
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- CA2973961A1 CA2973961A1 CA2973961A CA2973961A CA2973961A1 CA 2973961 A1 CA2973961 A1 CA 2973961A1 CA 2973961 A CA2973961 A CA 2973961A CA 2973961 A CA2973961 A CA 2973961A CA 2973961 A1 CA2973961 A1 CA 2973961A1
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/22—Improving land use; Improving water use or availability; Controlling erosion
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
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Abstract
A method of producing an environmental modification product, the method comprising: preparing an aqueous mixture comprising a starting material including one or more microorganisms, at least one starch and water; placing the aqueous mixture in a microorganism growth environment for a first time interval; adding at least one sugar to the mixture; placing the mixture in a fermentation environment for a second time interval; adding a carrier, and a volume of oxygen to the aqueous mixture for a third time interval; separating the carrier from the mixture; and drying the carrier to obtain the environmental modification product. Where a soil inoculant is being produced, the carrier may include a zeolite and the one or more microorganisms may include mycorrhizal fungi. Where a remediation product is being produced, the one or more microorganisms may include one or more bacteria.
Description
SOIL INOCULANT AND METHOD OF PRODUCING SAME
Field:
The present disclosure relates to environmental modification products, including soil inoculants and soil remediation products, for inoculating or remediating soil and methods for producing same; in particular, the present disclosure relates to environmental modification products for introducing fungus or bacteria to a soil or other environment.
Background:
In cultivating plants or trees, it is known that soil additives, commonly referred to as soil inoculants, may be used to introduce beneficial moulds to the soil or growing medium of the plants, thereby promoting the growth and health of the plants being cultivated in that soil or growing medium.
In particular, a class of fungi known as mycorrhizal fungi, is a fungi which colonizes the plant's root tissues and exists by receiving sugar from the plants in exchange for moisture and nutrients gathered from the soil or growing medium by the fungal strands. Advantageously for the plant, the mycorrhizal fungi form a symbiotic relationship with the roots of the plant and effectively extends the surface area of the plant's root systems, thereby increasing the absorption of water and nutrients from the soil. As well, once the plant's roots are colonized by the mycorrhizal fungi, the plant and mycorrhizal fungi may work together to modify the pH of the surrounding soil so as to become more favourable for the colonized plant. Additionally, for unfavourable growing mediums such as acidic or contaminated soils, the colonization of the plant's roots by another class of fungi known as Trichoderma fungi may provide a protective function of the plant's roots by forming a protective barrier between the roots of the plant and the contaminants in the soil.
The beneficial mycorrhizal and/or Trichoderma fungi may be naturally present in certain growing media. However, when it is not present in a particular growing media, it may be desirable to amend that growing media so as to introduce the mycorrhizal and/or Trichoderma fungi to the growing media, for example by using a soil inoculant containing mycorrhizal and/or Trichoderma fungi.
Various different types of soil inoculants are available in the market, produced by various processes, some of which include first cultivating mycorrhizal fungi on a host plant, and then harvesting the mycorrhizal fungi from the host plant and processing it to thereby incorporate it into a soil inoculant C3203834.DOCX;1 1 product. For example, in patent application number PCT/ZA2008/000060 by inventor Venter (the' 060 patent application), a method of producing endomycorrhiza inoculum involves cultivating endomycorrhiza spores on roots of a host plant in a growing medium and sequences of steps to separate the spores and producing a spore concentrate on a carrier.
Specifically, the process embodiments described in the '060 patent application include removing a host plant from a growth medium and processing the growth medium containing spores and hyphae by washing it thoroughly with water or aqueous liquid, separating any root matter and coarse debris from the spore-containing liquid, separating the spores from the liquid, and mixing the spores with a fine absorbent carrier powder to obtain an nearly dry powder mixture. The carrier powder may include a fine zeolite powder and/or one or more clay minerals.
In patent application number PCT/ZA02/00046 by inventors Hilditch et al (the '046 patent application), a method of preparing a growth substrate for use in plant inoculation includes the steps of obtaining a viable source of microorganism, growing the microorganism in proximity to one or more propagative host plant root systems so as to encourage development of a symbiotic association between the microorganism and the propagative host plant root systems, supporting the microorganism and the propagative host plant root systems in a support medium being a suitable zeolite such as capezeo, allowing the microorganism to sporulate, terminating the growth of the propagative host plant root systems, allowing the microorganism tempter state of dormancy in response to the termination of growth of the propagative host plant root systems, and separating the microorganism, associated propagative host plant root systems and supports medium from the remainder of the propagative host plant.
In US patent number 8,728,460 by inventor Spittle (the '460 patent), a soil treatment composition is described which includes combining beneficial soil fungi and bacteria in a growth promoting nutrient medium and embedding it in an organic porous ceramic particle for direct delivery during soil aerification. The process of manufacturing the soil treatment composition, as described in the '460 patent, includes spraying the porous carrier particles with a biological soil treatment composition. Carbohydrates and other food sources for the dormant bacteria and fungi are included to increase the colony forming of the organisms.
In European patent application number 93913523.2 by inventor Sakai et al (the '523 patent application), a process for producing a substance inoculated with a vesicular arbuscular mycorrhizae (VAM) fungus is disclosed, which comprises cultivating a plant infected with a VAM fungus belonging to C3203834.DOCX;1 2 the genus Gigaspora by using a base material comprising calcined amber loam, or a mixture of calcined amber loam and calcined attapulgite, to thereby proliferate the VAM fungus.
The process prepares a substance inoculated with a VAM fungus. Examples of carriers may include, for example, zeolite, foam clay, talc, pearlite, vermiculite, calcined amber loam, pumice, limestone, soil, sand, coke and peat moss.
So as to prevent contamination of the carrier with indigenous microbes, the carrier is subjected to sterilization (including calcination) prior to its use. A sterilized soil or a calcined amber loam is preferably used. After the spore density has come to be sufficiently high, the use calcined amber loam or other carrier is recovered to isolate the formed VAM inoculant, which is optionally dried if desired to obtain the final inoculant product. In the examples given in the '523 patent application, the host plants are grown for a period of 16 weeks after transplantation before the soil inoculants were produced from the used calcined amber loam or other carriers.
Summary:
In accordance with the present disclosure, in one embodiment, a method for producing a soil inoculant containing mycorrhizal fungi as well as other micronutrients includes using a starting material, for example a forest humus having sufficient amounts of mycorrhizal fungi and, optionally, a desired mixture of micronutrients. The starting material is mixed together with water and an appropriate starchy carbohydrate so as to facilitate the growth of the mycorrhizal fungi.
The aqueous starting material/carbohydrate mixture is left in the dark for approximately one week.
Sugar or molasses are then added to the mixture and the mixture is again placed in the dark for another week allowing it to ferment, which turns the mixture into a water soluble, viscous mixture. A
carrier, such as zeolite, is then added along with oxygen for approximately 36 hours, causing the mycorrhizal fungi and/or Trichoderma fungi, as well as any available micronutrients, to become absorbed and/or adsorbed into the pores of the zeolite, and then the zeolite is separated from the aqueous mixture water and the zeolite is dried, preferably in the dark, to produce the soil inoculant product.
Generally speaking, soil amendment products may be used to add beneficial microorganisms, such as mycorrhizal fungi or Trichoderma fungi or yeast, and/or micronutrients, to a soil or growth medium (the terms growth medium and soil are used interchangeably herein), while soil remediation products or remediation products may be used to remove contaminants from a soil, growing medium or other environment, including but not limited to bodies of water or oil wells or gas wells.
C3203834.DOCX;1 3 In another aspect of the present disclosure, customized environmental modification products (a generic term encompassing various types of products, including but not limited to amendment products, soil inoculants, soil amendment products, soil remediation products or remediation products as referred to herein) may be produced for particular applications. For example, certain species of mycorrhizal fungi may be particularly beneficial to certain types of plants, and so a soil inoculant product may be customized to provide those particular mycorrhizal fungi species, in some embodiments also producing a particular ratio of those species, as well as customizing the particular types of micronutrients in accordance with the particular plant to be grown in the inoculated growth medium or soil.
In other aspects of the present disclosure, customized remediation products may be produced using the processes described herein. Remediation products may be produced which contain various types of bacteria which are capable of consuming various types of contaminants in a given soil, water, or wellhead or well environment. For example, soil inoculants containing E. Coli bacteria may be used to remediate soil which is contaminated from mine tailings or other industrial processes. Other examples include inoculants or remediation products which contain oleispira antarctica bacteria for consuming oil spills in cold water, alcaninvorax borkumensis bacteria for consuming oil spills in warm water, or nitrate-reducing bacteria which may consume hydrogen sulphide in sour gas wellheads.
In some embodiments of the present disclosure, a method of producing an environmental modification product is provided, the method comprising: preparing an aqueous mixture comprising a starting material including one or more microorganisms, at least one starch and water; placing the mixture in a microorganism growth environment for a first time interval;
adding at least one sugar to the mixture; placing the mixture in a fermentation environment for a second time interval; adding a carrier and a volume of oxygen to the aqueous mixture for a third time interval;
separating the carrier from the mixture; and drying the carrier to obtain the remediation product.
Where a soil inoculant is being produced, the carrier may include a zeolite and the one or more microorganisms may include mycrorrhizae. Where a remediation product is being produced, the carrier may include other types of carriers, including but not limited to zeolite, clay, talc, charcoal, or any combination of these or other suitable carriers.
C3203834.DOCX4 4 Detailed Description The methods and processes disclosed herein provide for the preparation and customization of soil inoculants or remediation products for various different applications. In one aspect of the present disclosure, a soil inoculant may be prepared containing one or more species of mycorrhizal fungi which .. promotes the growth of plants and trees by forming a symbiotic relationship between the fungi and the roots of the plants or trees underneath the soil. Optionally, other types of beneficial microorganisms for promoting plant growth may also be incorporated into the soil inoculant. For example, Trichoderma fungi or yeast may be incorporated into the soil inoculant, or other types of beneficial fungi or yeasts may be incorporated.
As is known, mycorrhizal fungi promotes the healthy growth and development of plants and trees in soil, or other growth media, and is naturally present in soils with high organic material content.
(The terms soil and growth medium are used interchangeably in the present disclosure). However, in agricultural and other environments the soil may be depleted of the naturally occurring mycorrhizal fungi over time, thereby requiring reintroduction of the mycorrhizal fungi to the soil so as to promote healthy plant growth. Furthermore, micronutrients are also essential to the healthy growth and development of various plants and trees. Similarly, while such micronutrients are often found in the natural environment, they may be depleted in certain areas over time where the soil has been used many times for growing plants or trees, for example on agricultural plots or in residential areas, and also thereby may require the addition or re-introduction of such micronutrients into the soil being used for growing plants.
In other prior art processes for producing soil inoculants containing mycorrhizal fungi, of which the applicant is aware, such processes usually involve cultivating mycorrhizal fungi on a host plant, typically for a period of several weeks, and then harvesting the mycorrhizal spores and other propagules from the host plant and processing the mycorrhizal propagules to produce the soil inoculant. In the processes and methods disclosed herein, by identifying a starting material which contains the mycorrhizal fungi and which is preferably void of contaminants, such as harmful bacteria, in the starting material, the applicant has discovered that there are processes by which the mycorrhizal fungi may be cultivated directly from the starting material, without having to use host plants to cultivate the mycorrhizal fungi over a period of weeks. Advantageously, the processes disclosed herein may therefore cost less and take less time to produce the soil inoculants with desired concentrations of mycorrhizal fungi for use in soil mycorrhizal reintroduction. Furthermore, the applicant has found that C3203834.DOCX;1 5 the processes disclosed herein require fewer steps, in comparison to other processes, because it is not necessary to first separate spores from the host plants before continuing to process the fungus propogules in order to produce the soil inoculant. Further advantageously, in some embodiments of the present disclosure, it may be the case that the mycorrhizal fungi concentration of the soil inoculants produced in the present processes disclosed herein may exceed the concentrations produced using other methods.
In other aspects of the present disclosure, where it is desired to add micronutrients to the soil inoculants, the careful selection of an appropriate starting material which contain those micronutrients may result in soil inoculants including concentrations of those desired micronutrients. Therefore, in some embodiments of the present disclosure, it is not necessary to provide an additional source of micronutrients in the manufacture of the soil inoculant because an appropriately selected starting material may already include those desired micronutrients. In other embodiments of the present disclosure, where the starting material does not contain the desired micronutrients that need to be added to the soil inoculant, or does not include the desired micronutrients in the required quantities, such micronutrients may simply be introduced into the mixture during the manufacture process of the soil inoculants, or may also be included in the selected zeolite to be added during the process. The addition of micronutrients may occur at any step in the process of producing the soil inoculant.
In other aspects of the present disclosure, in addition to creating soil inoculants for introducing mycorrhizal fungi and optionally, micronutrients to the soil, other types of remediation products may be produced for different purposes, such as for remediation purposes, using the same processes described herein. Again, this would involve the appropriate selection of the correct starting material required for creating such remediation products. As an example, without intending to be limiting, remediation products containing E. coil bacteria may be produced for the purpose of remediating soil having oil or petroleum contaminants which may be consumed by the E. coil bacteria. In that instance, an appropriate startihg material may include excrement, manure or other organic material containing E.
coli bacteria.
Other possible applications for the processes described herein include loading a carrier, such as zeolite, with other types of bacteria for remediating contaminated water, or for removing sour gas from sour gas wells or other types of wellheads. For example, without intending to be limiting, remediation products which contain oleispira antarctica bacteria may be used for consuming oil spills in cold water, alcaninvorax borkumensis bacteria for consuming oil spills in warm water, or nitrate-reducing bacteria C3203834.DOCX;1 6 which may consume hydrogen sulphide in sour gas wellheads or other types of wellheads. Without intending to be limiting, appropriate starting materials for producing a remediation product containing nitrate-reducing bacteria may include, for example, chicken manure. It will be appreciated by a person skilled in the art that various types of organic materials containing specific types of bacteria may be used as starting materials to produce soil inoculants or remediation products.
As mentioned, the processes disclosed herein for producing soil inoculants include identifying appropriate starting materials. Without intending to be limiting, in some embodiments of the present disclosure it is desirable to select starting materials which do not contain undesirable contaminants, for example harmful bacteria which may enter plants being grown for food and thereby enter the human .. food supply chain.
An example of an appropriate starting material, for soil inoculants designed to reintroduce mycorrhizal fungi into the soil for promoting plant growth, may include a compost or humus which contains the desired species of mycorrhizal fungi. Additionally, such starting materials may also include certain micronutrients. Compost or humus may often contain different mixtures of micronutrients which are desirable for promoting healthy plant growth for particular types of plants.
For example, without intending to be limiting, the applicant has sourced forest humus which contains a desirable mixture of different types of mycorrhizal fungi, and also includes various different micronutrients including in particular calcium, magnesium, copper, zinc, manganese and iron in quantities which are optimal for promoting healthy plant growth. In addition, the forest humus starting material identified by the applicant further includes smaller quantities of available nitrogen, phosphorus, potassium, and boron which again support the healthy growth of plants. Importantly, the forest humus source identified by the applicant for producing mycorrhizal fungi soil inoculant does not contain harmful bacteria or plant disease pathogens, according to analyses conducted on the starting material. In another analysis conducted on the starting material identified by the applicant, measurement of the fungal biomass indicates that the ratio of fungus to bacteria is approximately 2:1, indicating that the starting material is well structured with ample room for oxygen to circulate and for water to be stored. The presence of fungi in this starting material also indicates the minerals and nutrients are able to be stored and locked into the soil.
While the above description of the specific starting material, comprising forest humus identified by the applicant ,is an example of an appropriate starting material for the soil inoculant production C3203834.DOCX;1 7 disclosed herein, it will be appreciated by a person skilled in the art that other appropriate starting materials may not contain these exact components, may include other components, and are intended to be included in the scope of the present disclosure. For example, other appropriate starting materials may contain different mixtures of micronutrients, or may not have any micronutrients or negligible amounts of micronutrients. Other examples of compounds or substances in a starting material that may be useful include hunnic acid, which may increase the absorption of the microorganisms by the zeolite, in the processes described below.
Once an appropriate starting material has been identified, the process for producing a soil inoculant, in one aspect of the present disclosure, includes preparing an aqueous mixture comprising the identified starting material, at least one starch, and water. The aqueous mixture is then placed into a microorganism growth environment for a given time interval so as to facilitate the growth and multiplication of the microorganisms which exist in the starting material. For example, for producing a soil inoculant which includes mycorrhizal fungi, the aqueous mixture would include a starting material which has mycorrhizal fungi, such as the forest humus described above, and the volume of forest humus is then mixed with water and a starch so as to create an aqueous mixture that will promote the growth of the mycorrhizal fungi. Without intending to be limiting, an example for the ratio of components in the aqueous mixture would be approximately 80% humus (or other starting material), 10% water and 10% starchy carbohydrates by volume.
The starch provides a food source for the mycorrhizal fungi (or other microorganisms, in the .. case of producing remediation products) to grow. Appropriate starches may include, for example, oats, rice, but may include any other type of starch, including barley, grains, potato meal, cornstarch, coconut husks, peat, wood chips, corn or any other appropriate starch for promoting the growth of mycorrhizal fungi or other targeted microorganism, such as bacteria. Another starch source may include brewery waste, otherwise referred to as spent grain or leftover beer mash. Optionally, the starch may be ground up so as to increase the surface area of the starch in the aqueous mixture for the mycorrhizal fungi to feed upon. In other embodiments in which a remediation product is produced, the food source for a remediation product containing nitrate-reducing bacteria may include chicken manure or other appropriate materials containing nitrate for the nitrate-reducing bacteria to feed upon.
Once the aqueous mixture has been prepared, it is placed in a microorganism growth environment which promotes the growth of the mycorrhizal fungi or other microorganism that is desired to be produced. For example, in the case of mycorrhizal fungi, the applicant has found that C3203834.DOCX;1 8 placing the aqueous mixture into a dark environment in which there is little or no light, combined with maintaining a temperature in that environment of approximately 10 C for a first time interval of approximately one week effectively enables the mycorrhizal fungi to multiply to the required concentrations for producing the soil inoculant. However, it will be appreciated by a person skilled in .. the art that other microorganism growth environments may also be appropriate for promoting the growth of mycorrhizal fungi or other targeted microorganisms. For example, the temperature may vary, in some embodiments for promoting growth of mycorrhizal fungi or other fungi, between 5 C and 20 C, depending on the particular types of fungus being grown. In other embodiments, for example for producing remedial products containing bacteria, the temperature range may be between 15 C and 40 C, depending on the type of bacteria being grown.
As another example, the time interval for the microorganism's growth need not be limited to seven days, and for example may be approximately in the range of four to seven days. The specific growing conditions for promoting microorganism growth may vary depending on the type of microorganisms which are being cultivated for the soil inoculant or remediation product. However, in .. the applicant's experience, in the embodiments for producing a soil inoculant containing mycorrhizal fungi, leaving the aqueous mixture in a dark environment at a temperature of approximately 10 C for approximately seven days has produced sufficient mycorrhizal fungi growth for further processing to produce a mycorrhizal soil inoculant.
Once the aqueous solution contains sufficient amounts of mycorrhizal fungi, or other targeted .. microorganism, at least one sugar is added to the aqueous mixture, and then the aqueous mixture is replaced into an environment adjusted for fermentation purposes for a second time interval. For example, without intending to be limiting, sugar may be introduced to the aqueous mixture at a ratio of one part sugar to two parts aqueous solution, by volume. The addition of sugar to the aqueous mixture enables fermentation of the mixture. The applicant has found that this process produces a substantially .. homogenous, viscous mixture, without chunks of humus or other starting material. Any type of sugar may be used and is intended to be included in the scope of the present disclosure. Without intending to be limiting, different types of sugars which may be added include cane sugar, beet sugar, molasses, or other appropriate types of sugar for encouraging fermentation. The fermentation promoting environment may include, for an example, placing the aqueous mixture with the sugar added into a dark .. environment in which there is little or no light, and maintaining the temperature of the environment in the range of 5 C to 20 C, and preferably, in the range of approximately 10 C, for embodiments to C3203834.DOCX;1 9 produce soil inoculants containing mycorrhizal fungi. The second time interval, for example without intending to be limiting, may be approximately one to two weeks. In other embodiments for producing remedial products containing bacteria, the environment may be maintained at a temperature in the range of 15 C to 40 C.
In addition to facilitating fermentation, the applicant suspects the addition of sugar at this stage in the process may play a further role in extending the shelf life of the final soil inoculant product, by providing a food source for the mycorrhizal fungi (or other targeted microorganism). For example, without intending to be limiting, the shelf life of the final soil inoculant product may be in the approximate range of one to two years.
Once the fermentation stage has completed, a carrier, such as a zeolite, and a volume of oxygen is added to the aqueous mixture, the oxygen being added for example by means of a bubbler. The oxygen promotes the absorption of the mycorrhizal fungi and of the micronutrients into the pores of the zeolite. The applicant has found, in some aspects of the present disclosure, that the selection of an appropriate zeolite will affect the final characteristics of the soil inoculant product. Zeolites are alumina silica structures which provide a rigid crystalline network, whereby zeolite particles include pores and therefore have a large surface area compared to other types of crystalline structures. Advantageously, the zeolites having a large surface area are able to absorb mycorrhizal spores and other mycorrhizal propagules, as well as micronutrients, from the aqueous solution. An additional benefit of zeolites is that they also absorb water molecules into their pores and release the water molecules overtime, which improves the water retention properties of the soil being treated with the soil inoculant.
There are many different types of zeolites having different crystalline structures and varying pore sizes, which may make certain zeolites more useful for this application than others zeolites given their particular affinities for certain ionic micronutrients and/or the mycorrhizal propagules.
Furthermore, certain zeolites may further advantageously have certain micronutrients, such as iron, already absorbed into the zeolite structure prior to processing in the methods described herein.
Without intending to be limiting, two examples of zeolites that may be appropriate for creating mycorrhizal-containing soil inoculants include chabazite, which contains available iron, and clinoptilolite.
Advantageously, both chabazite and clinoptilolite are capable of absorbing or adsorbing microorganisms, micronutrients and/or water molecules. However, it will be appreciated by person .. skilled in the art that other types of zeolites may also work, depending on the desired characteristics of C3203834.DOCX4 10 the final end product of the soil inoculant or remediation product, and that such types of zeolites are intended to be included in the scope of the present disclosure.
Depending on the particular environment to be remediated, such as a body of water, carriers other than zeolite, or in addition to zeolite, may be used in the production of a remediation product.
For example, for remediating a body of water, a carrier which floats or which may be suspended in water may be selected. Examples of carriers other than zeolite may include different types of clays, talc powders, charcoals or a combination of any of these carriers.
During this stage of the process, the zeolite is combined with the aqueous mixture at a ratio, for example, of approximately one part aqueous mixture, ten parts zeolite and ten parts water by volume.
The addition of oxygen to the mixture may assist in facilitating the absorption of the microorganisms, such as the mycorrhizal fungi, and the micronutrients into the zeolites pores.
The oxygen may also promote anaerobic fungal and bacterial growth. In some embodiments, the oxygen may be bubbled through the mixture or a period of substantially 36 hours. Preferably, this oxygenation step in the process will occur in a dark and temperature controlled environment; for example, in some embodiments the environment will be dark with little or no UV light in the environment, and the temperature may be maintained in the range of 5 C to 20 C, for example at a temperature of substantially 10 C. However, it will be appreciated by person skilled in the art that other time intervals for the oxygenation process and other environments may also be appropriate and are included within the present scope of this disclosure.
Once the zeolite has absorbed and/or adsorbed a sufficient amount of mycorrhizal fungi and micronutrients, the zeolite may be separated from the aqueous mixture, for example by filtering zeolite mixture through a sieve or screen. The recovered zeolite is then dried so as to remove excess water and obtain the final soil inoculant product. The drying process may occur, for example, in the dark (or in the absence of light in the UV spectrum), with dehumidifiers, for a period of time, which may take up to three days for example. Preferably, the drying process occurs in a temperature controlled environment, for example in the range of 15 C. It will be appreciated by a person skilled in the art that other drying procedures may be used and are intended to be included in the present scope of this disclosure. The aqueous mixture that is separated from the zeolite may optionally be used as a liquid soil inoculant product, as a certain amount of micronutrients and/or mycorrhizal fungi propagules may remain within the aqueous solution.
C3203834.DOCX;1 11 As discussed above, other types of environmental modification products, such as remediation products, may also be produced by the procedures described above. In some embodiments, where it is desirable to produce a remediation product containing bacteria for use in soil reclamation projects, the remediation product may be prepared by selecting an appropriate starting material which contains the target bacteria to be incorporated into the remediation product. Examples stated above include cultivating an E. coli bacteria by using a starting material which is an organic material containing an amount of E. coli bacteria. Examples of such starting materials might include for example excrement or manure, or any other organic materials that are contaminated by E. coli. These examples are not intended to be limiting, and it will be appreciated by person skilled in the art that other types of bacteria may also be produced in certain remediation products by selecting other starting materials which contain the desired targeted bacteria.
Example 1 To illustrate the process of producing a soil inoculant in accordance with the present disclosure, without intending to be limiting, the applicant utilizes a starting material that is forest humus containing various types of mycorrhizal fungi, as well as micronutrients including calcium, magnesium, copper, zinc, manganese, boron, sulphate, nitrogen, phosphorus, and potassium. In this example, one metric tonne of starting material may be used to produce approximately 20 metric tonnes of soil inoculant.
To begin the process, an aqueous solution was prepared by mixing approximately eight parts of the humus starting material with one part water and one part oats by volume.
Once thoroughly mixed, the aqueous solution was placed in a dark, temperature controlled environment, where the temperature was maintained at approximately 10 C for a period of approximately one week. At the end of this first time interval, a volume of brown sugar was mixed into the aqueous solution at a ratio of one part sugar to two parts aqueous solution, and then the solution was then replaced in the temperature controlled, darkened environment wherein the temperature is maintained at approximately 10 C for a second time interval of approximately one week. At the end of the second time interval, during which fermentation of the aqueous solution has occurred, a volume of zeolite and water were mixed into the aqueous solution, at a ratio of one part aqueous solution to ten parts clinoptilolite zeolite and ten parts water, and then an oxygen bubbler was used to introduce oxygen to the mixture for a period of approximately 36 hours. Again, this process takes place in a dark, temperature controlled environment C3203834.DOCX;1 12 wherein the temperature was maintained at approximately 10 C for a period of 36 hours. Once the oxygenation process was completed, the mixture was poured through a sieve to separate the zeolite from the aqueous solution. In this example, the aqueous solution was retained to be used as a liquid soil inoculant. The zeolite separated from the aqueous solution was then dried by spreading it out on a .. clean surface and running a series of fans and dehumidifiers for approximately three days to obtain the final soil inoculant product. The zeolite was frequently turned to facilitate the drying process.
In use, the soil inoculant is typically mixed with a soil or growth medium and then applied to the area of interest. The ratios of soil inoculant to growth medium for the mixture may vary, depending on the application. For example, without intending to be limiting, for a raised garden bed application, the applicant would recommend using the final soil inoculant product by mixing one part (5%) inoculant to nineteen parts (95%) other soil or growth medium components, such as compost or coco fiber, whereby the mixture ratios are by volume. In the case of top dressing crops or gardens, one can mix the inoculant in a ratio of up to one part (20%) soil inoculant to four parts (80%) other soil ingredients, the soil inoculant mixture intended to be spread evenly over the area of interest.
Advantageously, using a soil inoculant produced in accordance with Example 1 above, the applicant would be able to inoculate approximately 200 metric tons of soil for a raised garden bed application or 80 metric tons of soil for crops or gardens, utilizing a soil inoculant produced from one metric ton of starting material.
In an analysis of the soil inoculant produced in Example 1 above, surprisingly high levels of yeast and mould, including the mycorrhizal fungi, were identified in the sample.
Specifically, one gram of soil inoculant was found to contain 1.0 x 107 colony forming units (cfu) of mould and yeast.
C3203834.DOCX;1 13
Field:
The present disclosure relates to environmental modification products, including soil inoculants and soil remediation products, for inoculating or remediating soil and methods for producing same; in particular, the present disclosure relates to environmental modification products for introducing fungus or bacteria to a soil or other environment.
Background:
In cultivating plants or trees, it is known that soil additives, commonly referred to as soil inoculants, may be used to introduce beneficial moulds to the soil or growing medium of the plants, thereby promoting the growth and health of the plants being cultivated in that soil or growing medium.
In particular, a class of fungi known as mycorrhizal fungi, is a fungi which colonizes the plant's root tissues and exists by receiving sugar from the plants in exchange for moisture and nutrients gathered from the soil or growing medium by the fungal strands. Advantageously for the plant, the mycorrhizal fungi form a symbiotic relationship with the roots of the plant and effectively extends the surface area of the plant's root systems, thereby increasing the absorption of water and nutrients from the soil. As well, once the plant's roots are colonized by the mycorrhizal fungi, the plant and mycorrhizal fungi may work together to modify the pH of the surrounding soil so as to become more favourable for the colonized plant. Additionally, for unfavourable growing mediums such as acidic or contaminated soils, the colonization of the plant's roots by another class of fungi known as Trichoderma fungi may provide a protective function of the plant's roots by forming a protective barrier between the roots of the plant and the contaminants in the soil.
The beneficial mycorrhizal and/or Trichoderma fungi may be naturally present in certain growing media. However, when it is not present in a particular growing media, it may be desirable to amend that growing media so as to introduce the mycorrhizal and/or Trichoderma fungi to the growing media, for example by using a soil inoculant containing mycorrhizal and/or Trichoderma fungi.
Various different types of soil inoculants are available in the market, produced by various processes, some of which include first cultivating mycorrhizal fungi on a host plant, and then harvesting the mycorrhizal fungi from the host plant and processing it to thereby incorporate it into a soil inoculant C3203834.DOCX;1 1 product. For example, in patent application number PCT/ZA2008/000060 by inventor Venter (the' 060 patent application), a method of producing endomycorrhiza inoculum involves cultivating endomycorrhiza spores on roots of a host plant in a growing medium and sequences of steps to separate the spores and producing a spore concentrate on a carrier.
Specifically, the process embodiments described in the '060 patent application include removing a host plant from a growth medium and processing the growth medium containing spores and hyphae by washing it thoroughly with water or aqueous liquid, separating any root matter and coarse debris from the spore-containing liquid, separating the spores from the liquid, and mixing the spores with a fine absorbent carrier powder to obtain an nearly dry powder mixture. The carrier powder may include a fine zeolite powder and/or one or more clay minerals.
In patent application number PCT/ZA02/00046 by inventors Hilditch et al (the '046 patent application), a method of preparing a growth substrate for use in plant inoculation includes the steps of obtaining a viable source of microorganism, growing the microorganism in proximity to one or more propagative host plant root systems so as to encourage development of a symbiotic association between the microorganism and the propagative host plant root systems, supporting the microorganism and the propagative host plant root systems in a support medium being a suitable zeolite such as capezeo, allowing the microorganism to sporulate, terminating the growth of the propagative host plant root systems, allowing the microorganism tempter state of dormancy in response to the termination of growth of the propagative host plant root systems, and separating the microorganism, associated propagative host plant root systems and supports medium from the remainder of the propagative host plant.
In US patent number 8,728,460 by inventor Spittle (the '460 patent), a soil treatment composition is described which includes combining beneficial soil fungi and bacteria in a growth promoting nutrient medium and embedding it in an organic porous ceramic particle for direct delivery during soil aerification. The process of manufacturing the soil treatment composition, as described in the '460 patent, includes spraying the porous carrier particles with a biological soil treatment composition. Carbohydrates and other food sources for the dormant bacteria and fungi are included to increase the colony forming of the organisms.
In European patent application number 93913523.2 by inventor Sakai et al (the '523 patent application), a process for producing a substance inoculated with a vesicular arbuscular mycorrhizae (VAM) fungus is disclosed, which comprises cultivating a plant infected with a VAM fungus belonging to C3203834.DOCX;1 2 the genus Gigaspora by using a base material comprising calcined amber loam, or a mixture of calcined amber loam and calcined attapulgite, to thereby proliferate the VAM fungus.
The process prepares a substance inoculated with a VAM fungus. Examples of carriers may include, for example, zeolite, foam clay, talc, pearlite, vermiculite, calcined amber loam, pumice, limestone, soil, sand, coke and peat moss.
So as to prevent contamination of the carrier with indigenous microbes, the carrier is subjected to sterilization (including calcination) prior to its use. A sterilized soil or a calcined amber loam is preferably used. After the spore density has come to be sufficiently high, the use calcined amber loam or other carrier is recovered to isolate the formed VAM inoculant, which is optionally dried if desired to obtain the final inoculant product. In the examples given in the '523 patent application, the host plants are grown for a period of 16 weeks after transplantation before the soil inoculants were produced from the used calcined amber loam or other carriers.
Summary:
In accordance with the present disclosure, in one embodiment, a method for producing a soil inoculant containing mycorrhizal fungi as well as other micronutrients includes using a starting material, for example a forest humus having sufficient amounts of mycorrhizal fungi and, optionally, a desired mixture of micronutrients. The starting material is mixed together with water and an appropriate starchy carbohydrate so as to facilitate the growth of the mycorrhizal fungi.
The aqueous starting material/carbohydrate mixture is left in the dark for approximately one week.
Sugar or molasses are then added to the mixture and the mixture is again placed in the dark for another week allowing it to ferment, which turns the mixture into a water soluble, viscous mixture. A
carrier, such as zeolite, is then added along with oxygen for approximately 36 hours, causing the mycorrhizal fungi and/or Trichoderma fungi, as well as any available micronutrients, to become absorbed and/or adsorbed into the pores of the zeolite, and then the zeolite is separated from the aqueous mixture water and the zeolite is dried, preferably in the dark, to produce the soil inoculant product.
Generally speaking, soil amendment products may be used to add beneficial microorganisms, such as mycorrhizal fungi or Trichoderma fungi or yeast, and/or micronutrients, to a soil or growth medium (the terms growth medium and soil are used interchangeably herein), while soil remediation products or remediation products may be used to remove contaminants from a soil, growing medium or other environment, including but not limited to bodies of water or oil wells or gas wells.
C3203834.DOCX;1 3 In another aspect of the present disclosure, customized environmental modification products (a generic term encompassing various types of products, including but not limited to amendment products, soil inoculants, soil amendment products, soil remediation products or remediation products as referred to herein) may be produced for particular applications. For example, certain species of mycorrhizal fungi may be particularly beneficial to certain types of plants, and so a soil inoculant product may be customized to provide those particular mycorrhizal fungi species, in some embodiments also producing a particular ratio of those species, as well as customizing the particular types of micronutrients in accordance with the particular plant to be grown in the inoculated growth medium or soil.
In other aspects of the present disclosure, customized remediation products may be produced using the processes described herein. Remediation products may be produced which contain various types of bacteria which are capable of consuming various types of contaminants in a given soil, water, or wellhead or well environment. For example, soil inoculants containing E. Coli bacteria may be used to remediate soil which is contaminated from mine tailings or other industrial processes. Other examples include inoculants or remediation products which contain oleispira antarctica bacteria for consuming oil spills in cold water, alcaninvorax borkumensis bacteria for consuming oil spills in warm water, or nitrate-reducing bacteria which may consume hydrogen sulphide in sour gas wellheads.
In some embodiments of the present disclosure, a method of producing an environmental modification product is provided, the method comprising: preparing an aqueous mixture comprising a starting material including one or more microorganisms, at least one starch and water; placing the mixture in a microorganism growth environment for a first time interval;
adding at least one sugar to the mixture; placing the mixture in a fermentation environment for a second time interval; adding a carrier and a volume of oxygen to the aqueous mixture for a third time interval;
separating the carrier from the mixture; and drying the carrier to obtain the remediation product.
Where a soil inoculant is being produced, the carrier may include a zeolite and the one or more microorganisms may include mycrorrhizae. Where a remediation product is being produced, the carrier may include other types of carriers, including but not limited to zeolite, clay, talc, charcoal, or any combination of these or other suitable carriers.
C3203834.DOCX4 4 Detailed Description The methods and processes disclosed herein provide for the preparation and customization of soil inoculants or remediation products for various different applications. In one aspect of the present disclosure, a soil inoculant may be prepared containing one or more species of mycorrhizal fungi which .. promotes the growth of plants and trees by forming a symbiotic relationship between the fungi and the roots of the plants or trees underneath the soil. Optionally, other types of beneficial microorganisms for promoting plant growth may also be incorporated into the soil inoculant. For example, Trichoderma fungi or yeast may be incorporated into the soil inoculant, or other types of beneficial fungi or yeasts may be incorporated.
As is known, mycorrhizal fungi promotes the healthy growth and development of plants and trees in soil, or other growth media, and is naturally present in soils with high organic material content.
(The terms soil and growth medium are used interchangeably in the present disclosure). However, in agricultural and other environments the soil may be depleted of the naturally occurring mycorrhizal fungi over time, thereby requiring reintroduction of the mycorrhizal fungi to the soil so as to promote healthy plant growth. Furthermore, micronutrients are also essential to the healthy growth and development of various plants and trees. Similarly, while such micronutrients are often found in the natural environment, they may be depleted in certain areas over time where the soil has been used many times for growing plants or trees, for example on agricultural plots or in residential areas, and also thereby may require the addition or re-introduction of such micronutrients into the soil being used for growing plants.
In other prior art processes for producing soil inoculants containing mycorrhizal fungi, of which the applicant is aware, such processes usually involve cultivating mycorrhizal fungi on a host plant, typically for a period of several weeks, and then harvesting the mycorrhizal spores and other propagules from the host plant and processing the mycorrhizal propagules to produce the soil inoculant. In the processes and methods disclosed herein, by identifying a starting material which contains the mycorrhizal fungi and which is preferably void of contaminants, such as harmful bacteria, in the starting material, the applicant has discovered that there are processes by which the mycorrhizal fungi may be cultivated directly from the starting material, without having to use host plants to cultivate the mycorrhizal fungi over a period of weeks. Advantageously, the processes disclosed herein may therefore cost less and take less time to produce the soil inoculants with desired concentrations of mycorrhizal fungi for use in soil mycorrhizal reintroduction. Furthermore, the applicant has found that C3203834.DOCX;1 5 the processes disclosed herein require fewer steps, in comparison to other processes, because it is not necessary to first separate spores from the host plants before continuing to process the fungus propogules in order to produce the soil inoculant. Further advantageously, in some embodiments of the present disclosure, it may be the case that the mycorrhizal fungi concentration of the soil inoculants produced in the present processes disclosed herein may exceed the concentrations produced using other methods.
In other aspects of the present disclosure, where it is desired to add micronutrients to the soil inoculants, the careful selection of an appropriate starting material which contain those micronutrients may result in soil inoculants including concentrations of those desired micronutrients. Therefore, in some embodiments of the present disclosure, it is not necessary to provide an additional source of micronutrients in the manufacture of the soil inoculant because an appropriately selected starting material may already include those desired micronutrients. In other embodiments of the present disclosure, where the starting material does not contain the desired micronutrients that need to be added to the soil inoculant, or does not include the desired micronutrients in the required quantities, such micronutrients may simply be introduced into the mixture during the manufacture process of the soil inoculants, or may also be included in the selected zeolite to be added during the process. The addition of micronutrients may occur at any step in the process of producing the soil inoculant.
In other aspects of the present disclosure, in addition to creating soil inoculants for introducing mycorrhizal fungi and optionally, micronutrients to the soil, other types of remediation products may be produced for different purposes, such as for remediation purposes, using the same processes described herein. Again, this would involve the appropriate selection of the correct starting material required for creating such remediation products. As an example, without intending to be limiting, remediation products containing E. coil bacteria may be produced for the purpose of remediating soil having oil or petroleum contaminants which may be consumed by the E. coil bacteria. In that instance, an appropriate startihg material may include excrement, manure or other organic material containing E.
coli bacteria.
Other possible applications for the processes described herein include loading a carrier, such as zeolite, with other types of bacteria for remediating contaminated water, or for removing sour gas from sour gas wells or other types of wellheads. For example, without intending to be limiting, remediation products which contain oleispira antarctica bacteria may be used for consuming oil spills in cold water, alcaninvorax borkumensis bacteria for consuming oil spills in warm water, or nitrate-reducing bacteria C3203834.DOCX;1 6 which may consume hydrogen sulphide in sour gas wellheads or other types of wellheads. Without intending to be limiting, appropriate starting materials for producing a remediation product containing nitrate-reducing bacteria may include, for example, chicken manure. It will be appreciated by a person skilled in the art that various types of organic materials containing specific types of bacteria may be used as starting materials to produce soil inoculants or remediation products.
As mentioned, the processes disclosed herein for producing soil inoculants include identifying appropriate starting materials. Without intending to be limiting, in some embodiments of the present disclosure it is desirable to select starting materials which do not contain undesirable contaminants, for example harmful bacteria which may enter plants being grown for food and thereby enter the human .. food supply chain.
An example of an appropriate starting material, for soil inoculants designed to reintroduce mycorrhizal fungi into the soil for promoting plant growth, may include a compost or humus which contains the desired species of mycorrhizal fungi. Additionally, such starting materials may also include certain micronutrients. Compost or humus may often contain different mixtures of micronutrients which are desirable for promoting healthy plant growth for particular types of plants.
For example, without intending to be limiting, the applicant has sourced forest humus which contains a desirable mixture of different types of mycorrhizal fungi, and also includes various different micronutrients including in particular calcium, magnesium, copper, zinc, manganese and iron in quantities which are optimal for promoting healthy plant growth. In addition, the forest humus starting material identified by the applicant further includes smaller quantities of available nitrogen, phosphorus, potassium, and boron which again support the healthy growth of plants. Importantly, the forest humus source identified by the applicant for producing mycorrhizal fungi soil inoculant does not contain harmful bacteria or plant disease pathogens, according to analyses conducted on the starting material. In another analysis conducted on the starting material identified by the applicant, measurement of the fungal biomass indicates that the ratio of fungus to bacteria is approximately 2:1, indicating that the starting material is well structured with ample room for oxygen to circulate and for water to be stored. The presence of fungi in this starting material also indicates the minerals and nutrients are able to be stored and locked into the soil.
While the above description of the specific starting material, comprising forest humus identified by the applicant ,is an example of an appropriate starting material for the soil inoculant production C3203834.DOCX;1 7 disclosed herein, it will be appreciated by a person skilled in the art that other appropriate starting materials may not contain these exact components, may include other components, and are intended to be included in the scope of the present disclosure. For example, other appropriate starting materials may contain different mixtures of micronutrients, or may not have any micronutrients or negligible amounts of micronutrients. Other examples of compounds or substances in a starting material that may be useful include hunnic acid, which may increase the absorption of the microorganisms by the zeolite, in the processes described below.
Once an appropriate starting material has been identified, the process for producing a soil inoculant, in one aspect of the present disclosure, includes preparing an aqueous mixture comprising the identified starting material, at least one starch, and water. The aqueous mixture is then placed into a microorganism growth environment for a given time interval so as to facilitate the growth and multiplication of the microorganisms which exist in the starting material. For example, for producing a soil inoculant which includes mycorrhizal fungi, the aqueous mixture would include a starting material which has mycorrhizal fungi, such as the forest humus described above, and the volume of forest humus is then mixed with water and a starch so as to create an aqueous mixture that will promote the growth of the mycorrhizal fungi. Without intending to be limiting, an example for the ratio of components in the aqueous mixture would be approximately 80% humus (or other starting material), 10% water and 10% starchy carbohydrates by volume.
The starch provides a food source for the mycorrhizal fungi (or other microorganisms, in the .. case of producing remediation products) to grow. Appropriate starches may include, for example, oats, rice, but may include any other type of starch, including barley, grains, potato meal, cornstarch, coconut husks, peat, wood chips, corn or any other appropriate starch for promoting the growth of mycorrhizal fungi or other targeted microorganism, such as bacteria. Another starch source may include brewery waste, otherwise referred to as spent grain or leftover beer mash. Optionally, the starch may be ground up so as to increase the surface area of the starch in the aqueous mixture for the mycorrhizal fungi to feed upon. In other embodiments in which a remediation product is produced, the food source for a remediation product containing nitrate-reducing bacteria may include chicken manure or other appropriate materials containing nitrate for the nitrate-reducing bacteria to feed upon.
Once the aqueous mixture has been prepared, it is placed in a microorganism growth environment which promotes the growth of the mycorrhizal fungi or other microorganism that is desired to be produced. For example, in the case of mycorrhizal fungi, the applicant has found that C3203834.DOCX;1 8 placing the aqueous mixture into a dark environment in which there is little or no light, combined with maintaining a temperature in that environment of approximately 10 C for a first time interval of approximately one week effectively enables the mycorrhizal fungi to multiply to the required concentrations for producing the soil inoculant. However, it will be appreciated by a person skilled in .. the art that other microorganism growth environments may also be appropriate for promoting the growth of mycorrhizal fungi or other targeted microorganisms. For example, the temperature may vary, in some embodiments for promoting growth of mycorrhizal fungi or other fungi, between 5 C and 20 C, depending on the particular types of fungus being grown. In other embodiments, for example for producing remedial products containing bacteria, the temperature range may be between 15 C and 40 C, depending on the type of bacteria being grown.
As another example, the time interval for the microorganism's growth need not be limited to seven days, and for example may be approximately in the range of four to seven days. The specific growing conditions for promoting microorganism growth may vary depending on the type of microorganisms which are being cultivated for the soil inoculant or remediation product. However, in .. the applicant's experience, in the embodiments for producing a soil inoculant containing mycorrhizal fungi, leaving the aqueous mixture in a dark environment at a temperature of approximately 10 C for approximately seven days has produced sufficient mycorrhizal fungi growth for further processing to produce a mycorrhizal soil inoculant.
Once the aqueous solution contains sufficient amounts of mycorrhizal fungi, or other targeted .. microorganism, at least one sugar is added to the aqueous mixture, and then the aqueous mixture is replaced into an environment adjusted for fermentation purposes for a second time interval. For example, without intending to be limiting, sugar may be introduced to the aqueous mixture at a ratio of one part sugar to two parts aqueous solution, by volume. The addition of sugar to the aqueous mixture enables fermentation of the mixture. The applicant has found that this process produces a substantially .. homogenous, viscous mixture, without chunks of humus or other starting material. Any type of sugar may be used and is intended to be included in the scope of the present disclosure. Without intending to be limiting, different types of sugars which may be added include cane sugar, beet sugar, molasses, or other appropriate types of sugar for encouraging fermentation. The fermentation promoting environment may include, for an example, placing the aqueous mixture with the sugar added into a dark .. environment in which there is little or no light, and maintaining the temperature of the environment in the range of 5 C to 20 C, and preferably, in the range of approximately 10 C, for embodiments to C3203834.DOCX;1 9 produce soil inoculants containing mycorrhizal fungi. The second time interval, for example without intending to be limiting, may be approximately one to two weeks. In other embodiments for producing remedial products containing bacteria, the environment may be maintained at a temperature in the range of 15 C to 40 C.
In addition to facilitating fermentation, the applicant suspects the addition of sugar at this stage in the process may play a further role in extending the shelf life of the final soil inoculant product, by providing a food source for the mycorrhizal fungi (or other targeted microorganism). For example, without intending to be limiting, the shelf life of the final soil inoculant product may be in the approximate range of one to two years.
Once the fermentation stage has completed, a carrier, such as a zeolite, and a volume of oxygen is added to the aqueous mixture, the oxygen being added for example by means of a bubbler. The oxygen promotes the absorption of the mycorrhizal fungi and of the micronutrients into the pores of the zeolite. The applicant has found, in some aspects of the present disclosure, that the selection of an appropriate zeolite will affect the final characteristics of the soil inoculant product. Zeolites are alumina silica structures which provide a rigid crystalline network, whereby zeolite particles include pores and therefore have a large surface area compared to other types of crystalline structures. Advantageously, the zeolites having a large surface area are able to absorb mycorrhizal spores and other mycorrhizal propagules, as well as micronutrients, from the aqueous solution. An additional benefit of zeolites is that they also absorb water molecules into their pores and release the water molecules overtime, which improves the water retention properties of the soil being treated with the soil inoculant.
There are many different types of zeolites having different crystalline structures and varying pore sizes, which may make certain zeolites more useful for this application than others zeolites given their particular affinities for certain ionic micronutrients and/or the mycorrhizal propagules.
Furthermore, certain zeolites may further advantageously have certain micronutrients, such as iron, already absorbed into the zeolite structure prior to processing in the methods described herein.
Without intending to be limiting, two examples of zeolites that may be appropriate for creating mycorrhizal-containing soil inoculants include chabazite, which contains available iron, and clinoptilolite.
Advantageously, both chabazite and clinoptilolite are capable of absorbing or adsorbing microorganisms, micronutrients and/or water molecules. However, it will be appreciated by person .. skilled in the art that other types of zeolites may also work, depending on the desired characteristics of C3203834.DOCX4 10 the final end product of the soil inoculant or remediation product, and that such types of zeolites are intended to be included in the scope of the present disclosure.
Depending on the particular environment to be remediated, such as a body of water, carriers other than zeolite, or in addition to zeolite, may be used in the production of a remediation product.
For example, for remediating a body of water, a carrier which floats or which may be suspended in water may be selected. Examples of carriers other than zeolite may include different types of clays, talc powders, charcoals or a combination of any of these carriers.
During this stage of the process, the zeolite is combined with the aqueous mixture at a ratio, for example, of approximately one part aqueous mixture, ten parts zeolite and ten parts water by volume.
The addition of oxygen to the mixture may assist in facilitating the absorption of the microorganisms, such as the mycorrhizal fungi, and the micronutrients into the zeolites pores.
The oxygen may also promote anaerobic fungal and bacterial growth. In some embodiments, the oxygen may be bubbled through the mixture or a period of substantially 36 hours. Preferably, this oxygenation step in the process will occur in a dark and temperature controlled environment; for example, in some embodiments the environment will be dark with little or no UV light in the environment, and the temperature may be maintained in the range of 5 C to 20 C, for example at a temperature of substantially 10 C. However, it will be appreciated by person skilled in the art that other time intervals for the oxygenation process and other environments may also be appropriate and are included within the present scope of this disclosure.
Once the zeolite has absorbed and/or adsorbed a sufficient amount of mycorrhizal fungi and micronutrients, the zeolite may be separated from the aqueous mixture, for example by filtering zeolite mixture through a sieve or screen. The recovered zeolite is then dried so as to remove excess water and obtain the final soil inoculant product. The drying process may occur, for example, in the dark (or in the absence of light in the UV spectrum), with dehumidifiers, for a period of time, which may take up to three days for example. Preferably, the drying process occurs in a temperature controlled environment, for example in the range of 15 C. It will be appreciated by a person skilled in the art that other drying procedures may be used and are intended to be included in the present scope of this disclosure. The aqueous mixture that is separated from the zeolite may optionally be used as a liquid soil inoculant product, as a certain amount of micronutrients and/or mycorrhizal fungi propagules may remain within the aqueous solution.
C3203834.DOCX;1 11 As discussed above, other types of environmental modification products, such as remediation products, may also be produced by the procedures described above. In some embodiments, where it is desirable to produce a remediation product containing bacteria for use in soil reclamation projects, the remediation product may be prepared by selecting an appropriate starting material which contains the target bacteria to be incorporated into the remediation product. Examples stated above include cultivating an E. coli bacteria by using a starting material which is an organic material containing an amount of E. coli bacteria. Examples of such starting materials might include for example excrement or manure, or any other organic materials that are contaminated by E. coli. These examples are not intended to be limiting, and it will be appreciated by person skilled in the art that other types of bacteria may also be produced in certain remediation products by selecting other starting materials which contain the desired targeted bacteria.
Example 1 To illustrate the process of producing a soil inoculant in accordance with the present disclosure, without intending to be limiting, the applicant utilizes a starting material that is forest humus containing various types of mycorrhizal fungi, as well as micronutrients including calcium, magnesium, copper, zinc, manganese, boron, sulphate, nitrogen, phosphorus, and potassium. In this example, one metric tonne of starting material may be used to produce approximately 20 metric tonnes of soil inoculant.
To begin the process, an aqueous solution was prepared by mixing approximately eight parts of the humus starting material with one part water and one part oats by volume.
Once thoroughly mixed, the aqueous solution was placed in a dark, temperature controlled environment, where the temperature was maintained at approximately 10 C for a period of approximately one week. At the end of this first time interval, a volume of brown sugar was mixed into the aqueous solution at a ratio of one part sugar to two parts aqueous solution, and then the solution was then replaced in the temperature controlled, darkened environment wherein the temperature is maintained at approximately 10 C for a second time interval of approximately one week. At the end of the second time interval, during which fermentation of the aqueous solution has occurred, a volume of zeolite and water were mixed into the aqueous solution, at a ratio of one part aqueous solution to ten parts clinoptilolite zeolite and ten parts water, and then an oxygen bubbler was used to introduce oxygen to the mixture for a period of approximately 36 hours. Again, this process takes place in a dark, temperature controlled environment C3203834.DOCX;1 12 wherein the temperature was maintained at approximately 10 C for a period of 36 hours. Once the oxygenation process was completed, the mixture was poured through a sieve to separate the zeolite from the aqueous solution. In this example, the aqueous solution was retained to be used as a liquid soil inoculant. The zeolite separated from the aqueous solution was then dried by spreading it out on a .. clean surface and running a series of fans and dehumidifiers for approximately three days to obtain the final soil inoculant product. The zeolite was frequently turned to facilitate the drying process.
In use, the soil inoculant is typically mixed with a soil or growth medium and then applied to the area of interest. The ratios of soil inoculant to growth medium for the mixture may vary, depending on the application. For example, without intending to be limiting, for a raised garden bed application, the applicant would recommend using the final soil inoculant product by mixing one part (5%) inoculant to nineteen parts (95%) other soil or growth medium components, such as compost or coco fiber, whereby the mixture ratios are by volume. In the case of top dressing crops or gardens, one can mix the inoculant in a ratio of up to one part (20%) soil inoculant to four parts (80%) other soil ingredients, the soil inoculant mixture intended to be spread evenly over the area of interest.
Advantageously, using a soil inoculant produced in accordance with Example 1 above, the applicant would be able to inoculate approximately 200 metric tons of soil for a raised garden bed application or 80 metric tons of soil for crops or gardens, utilizing a soil inoculant produced from one metric ton of starting material.
In an analysis of the soil inoculant produced in Example 1 above, surprisingly high levels of yeast and mould, including the mycorrhizal fungi, were identified in the sample.
Specifically, one gram of soil inoculant was found to contain 1.0 x 107 colony forming units (cfu) of mould and yeast.
C3203834.DOCX;1 13
Claims (27)
1. A method of producing an environmental modification product, the method comprising:
preparing an aqueous mixture comprising a starting material including one or more microorganisms, at least one starch and water, placing the mixture in a microorganism growth environment for a first time interval so as to cultivate the one or more microorganisms, adding at least one sugar to the mixture, placing the mixture in a fermentation environment for a second time interval, adding a carrier and a volume of oxygen to the aqueous mixture for a third time interval, separating the carrier from the aqueous mixture, drying the carrier to obtain the environmental modification product.
preparing an aqueous mixture comprising a starting material including one or more microorganisms, at least one starch and water, placing the mixture in a microorganism growth environment for a first time interval so as to cultivate the one or more microorganisms, adding at least one sugar to the mixture, placing the mixture in a fermentation environment for a second time interval, adding a carrier and a volume of oxygen to the aqueous mixture for a third time interval, separating the carrier from the aqueous mixture, drying the carrier to obtain the environmental modification product.
2. The method of claim 1 wherein the environmental modification product is a soil inoculant for amending a soil and wherein the carrier includes a zeolite and the one or more microorganisms includes mycorrhizal fungi.
3. The method of claim 2 wherein the starting material includes humus.
4. The method of claim 3 wherein the starting material includes micronutrients.
5. The method of claim 1 wherein the one or more microorganisms includes one or more bacteria for consuming one or more contaminants.
6. The method of claim 5 wherein the starting material includes organic material containing E. Coli.
7. The method of claim 1 wherein the at least one starch is selected from a group comprising: rice, oats, barley, grains, coconut husks, peat, wood chips, corn, brewery waste.
8. The method of claim 1 wherein the at least one sugar is selected from a group comprising: cane sugar, brown sugar, molasses, beet sugar.
9. The method of claim 1 wherein the microorganism growth environment includes a dark environment having a temperature in the range of 5°C to 20°C.
10. The method of claim 9 wherein the temperature is substantially 10°C.
11. The method of claim 1 wherein the fermentation environment includes a dark environment having a temperature in the range of 5°C to 20°C.
12. The method of claim 11 wherein the temperature is substantially 10°C.
13. The method of claim 2 wherein the zeolite is selected from a group comprising: chabazite, clinoptilolite.
14. The method of claim 13 wherein the zeolite is characterized by having available micronutrients.
15. The method of claim 14 wherein the available micronutrients are selected from a group comprising: iron, calcium, boron, magnesium, zinc, manganese.
16. The method of claim 1 wherein the one or more microorganisms include bacteria selected from a group comprising: E. Coli, Oleispira Antarctica, Alcaninvorax Borkumensis, nitrate-reducing bacteria.
17. The method of claim 1 wherein the first time interval is selected from a range comprising substantially four days to seven days.
18. The method of claim 17 wherein the second time interval is selected from a range comprising seven days to fourteen days.
19. The method of claim 18 wherein the first and second time intervals are each substantially seven days.
20. The method of claim 19 wherein the third time interval is substantially 36 hours.
21. The method of claim 2, further comprising the step of using the aqueous mixture remaining after the zeolite has been separated from the mixture as a liquid soil inoculant.
22. An environmental modification product produced by the method of claim 1.
23. A soil inoculant produced by the method of claim 2.
24. The soil inoculant product of claim 23 wherein one gram of the the soil inoculant contains at least 1.0 x 10 7 colony forming units of a combination of yeast and fungi.
25. A method of producing an environmental modification product, the method consisting of the following steps:
preparing an aqueous mixture comprising a starting material including one or more microorganisms, at least one starch and water, placing the mixture in a microorganism growth environment for a first time interval so as to cultivate the one or more microorganisms, adding at least one sugar to the mixture, placing the mixture in a fermentation environment for a second time interval, adding a carrier and a volume of oxygen to the aqueous mixture for a third time interval, separating the carrier from the aqueous mixture, drying the carrier to obtain the environmental modification product.
preparing an aqueous mixture comprising a starting material including one or more microorganisms, at least one starch and water, placing the mixture in a microorganism growth environment for a first time interval so as to cultivate the one or more microorganisms, adding at least one sugar to the mixture, placing the mixture in a fermentation environment for a second time interval, adding a carrier and a volume of oxygen to the aqueous mixture for a third time interval, separating the carrier from the aqueous mixture, drying the carrier to obtain the environmental modification product.
26. The method of claim 25 wherein the environmental modification product is a soil inoculant for amending a soil and wherein the carrier includes a zeolite and the one or more microorganisms includes mycorrhizal fungi.
27. A soil inoculant produced by the method of claim 26.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2973961A CA2973961A1 (en) | 2017-07-20 | 2017-07-20 | Soil inoculant and method of producing same |
| CA2996590A CA2996590A1 (en) | 2017-07-20 | 2018-02-27 | Bio-stimulant and method of producing same |
| US16/040,048 US20190021339A1 (en) | 2017-07-20 | 2018-07-19 | Bio-stimulant and method of producing same |
| CA3011849A CA3011849C (en) | 2017-07-20 | 2018-07-19 | Bio-stimulant and method of producing same |
| US17/585,952 US20220144717A1 (en) | 2017-07-20 | 2022-01-27 | Bio-stimulant and method of producing same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2973961A CA2973961A1 (en) | 2017-07-20 | 2017-07-20 | Soil inoculant and method of producing same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2973961A1 true CA2973961A1 (en) | 2019-01-20 |
Family
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2973961A Abandoned CA2973961A1 (en) | 2017-07-20 | 2017-07-20 | Soil inoculant and method of producing same |
| CA2996590A Abandoned CA2996590A1 (en) | 2017-07-20 | 2018-02-27 | Bio-stimulant and method of producing same |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2996590A Abandoned CA2996590A1 (en) | 2017-07-20 | 2018-02-27 | Bio-stimulant and method of producing same |
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| CA (2) | CA2973961A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110903830A (en) * | 2019-12-23 | 2020-03-24 | 内蒙古科技大学 | Saline-alkali soil modifier and preparation method and application thereof |
| EP4111864A1 (en) | 2021-06-28 | 2023-01-04 | International Chemical Company S.A. | A bioformulation based on granulated oats hull and a method for its preparation |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110195028B (en) * | 2019-03-08 | 2022-03-01 | 大连理工大学 | Preparation method and application of compost low-temperature compound fermentation inoculant |
-
2017
- 2017-07-20 CA CA2973961A patent/CA2973961A1/en not_active Abandoned
-
2018
- 2018-02-27 CA CA2996590A patent/CA2996590A1/en not_active Abandoned
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110903830A (en) * | 2019-12-23 | 2020-03-24 | 内蒙古科技大学 | Saline-alkali soil modifier and preparation method and application thereof |
| EP4111864A1 (en) | 2021-06-28 | 2023-01-04 | International Chemical Company S.A. | A bioformulation based on granulated oats hull and a method for its preparation |
Also Published As
| Publication number | Publication date |
|---|---|
| CA2996590A1 (en) | 2019-01-20 |
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