CN113661234A

CN113661234A - Soil inoculation system based on microalgae and using method

Info

Publication number: CN113661234A
Application number: CN202080023859.8A
Authority: CN
Inventors: A·D·艾尔斯; M·R·爱德华兹; D·哈格; R·汤普森
Original assignee: Meilande Co ltd
Current assignee: Meilande Co ltd
Priority date: 2019-02-15
Filing date: 2020-02-14
Publication date: 2021-11-16
Also published as: IL285611A; CA3130211A1; MX2021009815A; WO2020168203A2; EP3924460A2; AU2020223331A1; EP3924460A4; PE20212119A1; CL2021002148A1; WO2020168203A3; BR112021016060A2; ECSP21067952A

Abstract

Some embodiments include a microalgae cultivation system including a bioreactor adapted to propagate microalgae in a culture broth using at least one of natural light and artificial light in combination with at least one nutrient including at least one carbon source, wherein the microalgae is freely suspended in and forms a portion of the culture broth. The microalgae feed source is coupled to the bioreactor and a first controller between the water conditioning assembly and the bioreactor. A water conditioning assembly is coupled to the bioreactor as an input for the supply water and is configured to condition the supply water to a specified purity that enables substantially unimpeded growth of microalgae in the broth to a specified concentration, and a first controller is configured to control the supply of the microalgae feed source to the bioreactor.

Description

Soil inoculation system based on microalgae and using method

RELATED APPLICATIONS

The present application claims the benefit and priority of U.S. provisional application No. 62/806,543 entitled "Method of Isolation, Selection, and Use of end microorganisms for agricultural Production Areas" filed on 15.2.2019. The present application claims priority from U.S. application No. 16/534,907, entitled "micro-Based Soil encapsulation System and Methods of Use", filed on 7.8.2019, which is a partial continuation of U.S. patent application No. 16/207,528, entitled "micro-Based Soil encapsulation System and Methods of Use", filed on 3.12.2018, 12.3, 16/207,528, which is a continuation of U.S. patent application No. 14/069,932 (now published as U.S. patent No. 10,172,304), filed on 1.11.2013, entitled "micro-Based Soil encapsulation System and Methods of Use", which U.S. patent No. 10,172,304 is an international patent application No. PCT/US 36293/38964 filed on 3.5.2012, entitled "micro-Based Soil encapsulation System and Methods of Use", the international patent application No. PCT/US12/36293 claims the benefit and priority of a partially-continued US provisional application No. 61/481,998 entitled "micro-Based Soil encapsulation Systems and Methods of Use" filed on 3.5.2011 and also U.S. patent application No. 15/647,005 entitled "Soil enhancement Systems and Methods" filed on 11.7.7.2017, which incorporates the disclosure of all such priority applications by reference. However, the present disclosure will be prioritized as it conflicts with any of the cited applications.

Background

Microorganisms in soil have many well-known beneficial effects. Although there are many references to algae herein, such references are used only as useful examples and do not limit the scope of the invention described and claimed herein, which also relates generally to microorganisms.

Algae have the ability to adapt to their environment. For example, algae found in southwest desert soils have been adapted to high temperatures, alkaline pH levels, and dry periods, while algae found in northern climates have been adapted to much lower temperatures, freeze-thaw cycles, higher soil moisture levels, and more acidic soil pH levels, among others.

The unique algae occupy ecological niches in the field ecosystem. In soil ecosystems, symbiosis with other organisms has been established, resulting in a biochemical environment where compounds produced by the characteristic algae may promote the growth of other desirable microorganisms and inhibit the growth of undesirable or non-beneficial organisms. For example, algae are known to produce biochemical substances such as amino acids, hormones, peptides, and fatty acids that promote the growth of beneficial microorganisms. These beneficial biochemical substances also directly aid the crop plants. Beneficial microorganisms produce biochemicals (e.g., sugars and vitamins) that algae and crops can utilize to grow, resulting in continued growth of algae and crops. At the same time, algae can produce antibacterial, antifungal, algicidal and/or antiprotozoal compounds that prevent the growth of harmful microorganisms in soil and surface water.

When the soil algae die, cellular biochemical is released, which can be directly supplied to the soil biofouling and any crop plants growing in the soil. These biochemical substances are large molecules (e.g., such as proteins, fats, dyes, peptides, nucleic acids, etc.), some or all of which can be absorbed by crop plants, thereby producing crops of greater nutritional value.

If live foreign algae are introduced into the soil, the ecosystem is forced to rebalance. This imbalance can result in the production of one or more undesirable biochemicals (such as toxins), or the lack of important biochemicals needed by the crop plant.

When algae are introduced into soil, the metabolic activity in the soil increases, resulting in more CO₂And (4) generating. This is particularly true for live algae, whose metabolic activity continues after entering the soil. This CO₂The generation of (2) reduces the pH of the soil, leads to the dissolution of calcium carbonate and magnesium carbonate bonds, thereby opening the soil, promoting the penetration of root systems and increasing the flow of water and fertilizer. This increased water movement brings more salt out of the root zone, thereby reducing the osmotic pressure within the root zone and increasing the bioavailability of macro and micronutrients to the crop. The lower pH also releases bound potassium and phosphorus, making it available to plants. Under phosphorus limiting conditions, algae secrete extracellular phosphatases almost immediately. These compounds release phosphates from soil particles, making them available to plants. Green algae also produce polysaccharides on which water can remain until it is needed.

The substantially constant or regular addition of algae can lead to the desired accumulation of organic matter (humus) in the soil, which also has the property of retaining moisture and nutrients that can be released to the plants as needed. Other methods of introducing humus into soil typically require turning the organic matter (compost, various plant cuttings, fertilizers, etc.), preferably between field planting. Humus contributes to the formation of the natural iron chelate (fulvic acid-iron), which prevents the soil from being clogged by calcium and magnesium carbonate, thus avoiding the problem of chlorosis due to the low bioavailability of these nutrients. Chlorosis is a reduction in the green color of plants due to a reduction in the amount of chlorophyll in the leaves due to the lack of bioavailable macro and micronutrients such as nitrogen (N), magnesium (Mg), calcium (Ca) and iron (Fe), even if these nutrients are present in the soil.

Ion exchange capacity is a quantitative means for describing the binding of fertilizer elements to soil particles for storage and release. The humus ion exchange capacity (e.g., 400 to 600 milliequivalents/100 g) is 5 to 10 times the ion exchange capacity (e.g., 50 to 150 milliequivalents/100 g) of the clay. It is this capacity that allows the fertilizer to remain in the soil for use by the plants as needed. When plants utilize nitrogen (N), phosphorus (P) and potassium (K) in soil, the stored elements are released from the humus when needed. By binding to humic substances, copper and other trace elements become less toxic and more readily available to plants.

If combined with microalgae, the fertilizer is more effective. Algal cells treat fertilizer by breaking down certain molecules into a bioavailable form that plants can more readily utilize. The nutrients are more efficiently and completely absorbed by the root system of the plant. For example, ammonium nitrate, a good source of nitrogen, is one of the most common bulk fertilizers (bulk ferizers) used to grow crops. Although the nitrate in such fertilizers can be immediately absorbed by the plant, the ammonium component is not readily available to the plant. Microalgae cells absorb ammonium, naturally convert it to nitrogenous biochemicals, and when they die, release these valuable biochemicals into plants for absorption and utilization. In addition, nutrients in the fertilizer can be combined with microalgae cells or their organic residues and are less likely to be lost to runoff water during rain or irrigation. When they die, algae can also nourish the soil for bacteria that can convert ammonium ions to nitrate ions.

Algae produce growth regulators (e.g., gibberellic acid) that can increase salt tolerance, induce seed germination, increase plant growth rate and fruit yield. Artificial or concentrated growth regulators are expensive, especially when used in large quantities, which makes it impossible for the grower to reproduce this effect by using other products.

Algae play a role in controlling agricultural pests by directly producing antibiotics and antifungal compounds, and by nourishing beneficial microorganisms in the soil that produce other anti-pest compounds. These compounds confer on plants the ability to prevent the invasion of pathogenic species. Diseases and pests are also resisted due to the improvement of plant vigor.

As mentioned above, live microalgae cells can act as catalysts, mining and utilizing all the benefits available from standard fertilizers; and also provides a natural supply of essential compounds and phytochemicals (phytochemicals) while supporting the overall efficacy of the growing environment. These powerful attributes work synergistically to stimulate healthier and faster plant growth; and continue to produce richer, higher quality, and more nutrient rich end products, such as crops. The benefits of the microalgae cell additive are available when the algal cells delivered to the soil are in a healthy living state and at high concentrations. The choice and formulation of the algae additive is critical to its overall impact. If properly formulated, microalgae additive programs are easy to manage and offer breakthrough potential in agricultural production. The effect is likely to be greatest in the most barren soils, such as arid soils with a high accumulation of salt and nitrate (caliche) and minimal organic matter. In addition, by selecting the specific algae for propagation and transporting the algae to an agricultural production area, the survival rate is higher, and the influence on the soil health is larger and faster.

Disclosure of Invention

Some embodiments include a culture system comprising a bioreactor adapted to propagate microalgae in a culture broth using natural and/or artificial light in combination with at least one nutrient comprising at least one carbon source, wherein the microalgae are freely suspended in the culture broth and form a portion of the culture broth. Some embodiments include an algae nutrient supply coupled to the bioreactor and a first controller between the water conditioning assembly and the bioreactor. In some embodiments, the water conditioning component is coupled to the bioreactor as an input for the water supply and is configured to condition the water supply to a specific purity such that microalgae in the culture broth can grow substantially unimpeded to a specified concentration. Additionally, in some embodiments, the first controller can be configured to control delivery of the algae nutrient supply to the bioreactor. In some embodiments, a carbon dioxide source is coupled to the bioreactor where carbon dioxide is injected into the culture fluid as a carbon source.

Some further embodiments include a second controller coupled to the probe and configured to regulate release of carbon dioxide from the carbon dioxide source to the bioreactor based at least in part on one or more measurements from the probe, wherein the carbon dioxide is injected into the culture fluid as the carbon source.

In some embodiments, the probe is a pH probe configured to measure the pH of the culture fluid. In some embodiments, the water conditioning assembly comprises an ozone generator coupled to the ozone contactor, wherein the ozone generator is configured to generate ozone and deliver the ozone to at least partially ozonate the feed water.

Some embodiments include a solids filter located downstream of the outlet of the ozone contactor, wherein the solids filter is configured to remove solids from ozonated feed water exiting the ozone contactor. Some embodiments include a carbon filter and/or an ultraviolet light system downstream of the solids filter, wherein the carbon filter and/or the ultraviolet light system can at least partially deodorize and oxidize the ozonated water supply.

Some embodiments include at least one source of pressurized air coupled to the bioreactor, wherein the at least one source of pressurized air is capable of generating bubbles to at least partially aerate and/or agitate the culture solution. In some embodiments, the gas bubbles comprise CO₂、N₂And/or O₂. Some embodiments further comprise at least one reservoir or tank that provides or is coupled to an input of a water supply.

Some further embodiments include a mobile cart supporting at least the bioreactor, the water conditioning assembly, and the carbon dioxide source. In some embodiments, the microalgae feed source comprises a fertilizer, macronutrients, micronutrients, and at least two different microalgae species.

In some embodiments, the macronutrient is selected from the group consisting of phosphorus, nitrogen, carbon, silicon, calcium salts, magnesium salts, sodium salts, potassium salts, and sulfur; the one or more micronutrients are selected from the group consisting of manganese, copper, zinc, cobalt, molybdenum, vitamins and trace elements. Additionally, in some embodiments, the micronutrients include vitamins and minerals that are added to the regulated water supply.

Some embodiments include a telemetry system configured to remotely monitor and/or control operation of one or more of the at least one component or assembly of the first controller, the second controller, the bioreactor and the water conditioning assembly.

In some embodiments, the artificial light comprises an LED lamp located within and/or near a bioreactor surface and exposing the microalgae to light.

In some embodiments, the carbon dioxide source comprises a tank containing carbon dioxide gas, and/or a carbon dioxide generator, and/or a carbon dioxide sequestration vessel that sequesters and temporarily stores atmospheric carbon dioxide.

In some further embodiments, the microalgae feed source comprises a first algae type and/or a second algae type, and/or bacteria and/or fungi. Some embodiments further include a flow imaging device coupled to an output of the bioreactor, wherein the flow imaging device is configured to generate images of algae, predators, and contaminants in the broth for quality control monitoring. Some embodiments further include a microbial mixer configured to blend algae and/or bacteria and/or fungi with any broth exiting the bioreactor.

Some embodiments include methods comprising preparing one or more microorganism-containing samples from at least one locus of a current or planned plant growth area, and preparing at least one cultured sample by culturing a microorganism from the sample. In addition, some embodiments include selecting at least one microbial target species from at least one culture sample, and propagating the at least one selected microbial target species to increase the concentration of the at least one microbial target species in the at least one culture sample. Some embodiments include providing a bioreactor adapted for propagating at least one selected target species in a culture broth, wherein the at least one selected target species is freely suspended in and forms a part of the culture broth. In addition, some embodiments include a feed source coupled to the bioreactor and a first controller between the water regulating assembly and the bioreactor, wherein the water regulating assembly is coupled to the bioreactor as an input to the water supply and is configured to regulate the water supply to a specified purity that allows for substantially unimpeded growth of at least one selected target species in the culture broth to a specified concentration. Additionally, in some embodiments, the first controller is configured to control the supply of the feed source to the bioreactor. Additionally, in some embodiments of the method, a carbon dioxide source is coupled to the bioreactor.

Some embodiments include a second controller coupled to the probe and configured to regulate release of carbon dioxide from the carbon dioxide source to the bioreactor based at least in part on one or more measurements from the probe, and further, the carbon source enables propagation of at least one selected microbial target species with the carbon dioxide injected into the culture broth. Some embodiments include delivering at least a portion of the at least one target microbial species to at least a portion of the at least one site, wherein the at least a portion of the at least one target microbial species delivered comprises at least one viable microorganism. In some embodiments, at least one viable microorganism is selected to be well-adapted to the proper species.

In some embodiments of the method, the water conditioning assembly comprises an ozone generator coupled to the ozone contactor, wherein the ozone generator is configured to generate ozone and deliver the ozone to at least partially ozonate the feed water.

In some embodiments of the method, a solids filter is placed upstream of the inlet of the ozone contactor.

In some embodiments of the method, a carbon filter and/or an ultraviolet light system is positioned directly downstream of the solids filter, wherein the carbon filter and/or the ultraviolet light system is configured and arranged to at least partially deodorize and oxidize the ozonated water supply. Additionally, at least one source of pressurized air is coupled to the bioreactor, wherein the at least one source of pressurized air can generate bubbles to at least partially aerate and/or agitate the culture solution in the bioreactor.

Some embodiments of the method further comprise transporting at least a portion of the at least one target microbial species to at least a portion of the at least one site. In some embodiments, the at least a portion of the delivered at least one target microbial species comprises at least one viable microorganism. In some embodiments, the at least one viable microorganism is an algal species characteristic of the delivery site. In some further embodiments, the at least one living microorganism is a living species selected to restore a normal soil flora combination in the field. In some other embodiments, the live species of algae are selected for their use in improving certain desired characteristics of the soil at the delivery site.

Some embodiments include methods comprising collecting a sample of an algal lineage from an agricultural locus, and selecting at least one desired algal species for propagation, wherein the at least one desired algal species is present in the agricultural locus at an initial concentration. Some embodiments include propagating at least one desired algae species in at least one bioreactor and delivering the at least one desired species to an agricultural site to increase the concentration of the algae species to a concentration greater than the initial concentration.

In some embodiments of the method, the at least one bioreactor is adapted to propagate at least one desired species in the culture broth using at least one of natural light and artificial light in combination with at least one nutrient comprising at least one carbon source, wherein the at least one desired species is freely suspended in and forms part of the culture broth.

In some embodiments of the method, an algae nutrient supply is coupled to the at least one bioreactor and a controller for controlling flow between the water conditioning assembly and the at least one bioreactor. In some embodiments, the water conditioning component is coupled to the at least one bioreactor as an input to the water supply to condition the water supply to a specified purity, which enables substantially unimpeded growth of microalgae in the culture broth to a specified concentration. Additionally, in some embodiments, the controller is configured to control the supply of algae nutrient to the at least one bioreactor.

In some embodiments of the method, a carbon dioxide source is coupled to the at least one bioreactor, wherein carbon dioxide is injected into the culture broth as the carbon source. In some further implementations of the method, a second controller is coupled to the probe, the second controller configured to regulate release of carbon dioxide from the carbon dioxide source to the bioreactor based at least in part on one or more measurements from the probe.

In some embodiments of the method, the water conditioning assembly comprises an ozone generator coupled to the ozone contactor, wherein the ozone generator generates ozone and delivers the ozone to at least partially ozonate the feed water. In some further embodiments of the method, the solids filter is located upstream of the inlet of the ozone contactor.

In some embodiments of the method, a carbon filter and/or an ultraviolet light system is located downstream of the solids filter, wherein at least one of the carbon filter and the ultraviolet light system at least partially deodorizes and oxidizes the ozonated water supply. In some other embodiments of the method, a pressurized air source is coupled to the bioreactor, wherein the pressurized air source generates air bubbles to at least partially aerate and/or agitate the culture fluid in the at least one bioreactor.

Drawings

Fig. 1 depicts a first embodiment of a microalgae-based soil inoculation system of the present invention.

FIG. 2 depicts a front perspective view of a second embodiment of a microalgae-based soil inoculation system of the present invention.

FIG. 3 depicts a side view of a third embodiment of a microalgae-based soil inoculation system of the present invention.

Fig. 4A depicts a field 5 weeks after planting and processing a melon crop according to the methods and systems of the present invention.

Fig. 4B depicts the same field of fig. 4A 9 weeks after planting and processing a melon crop according to the methods and systems of the present invention.

Fig. 5A depicts melon plants in a portion of a field that were not treated according to the present invention.

Fig. 5B depicts melon plants in a portion of a field treated according to the present invention.

Fig. 6A depicts melons after 9 weeks of planting in a portion of the field not treated according to the present invention.

Fig. 6B depicts melons after 9 weeks of planting in a portion of the field treated according to the present invention.

Fig. 7 depicts a fourth embodiment of a microalgae-based soil inoculation system of the present invention.

Fig. 8 depicts a fifth embodiment of a microalgae-based soil inoculation system of the present invention.

Fig. 9 illustrates a soil enrichment system according to some further embodiments of the present invention.

Detailed description of the preferred embodiments

Before any embodiments of the invention are explained in detail, it is to be understood that the invention is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the following drawings. The invention is capable of other embodiments and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of "including," "comprising," or "having" and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. Unless specified or limited otherwise, the terms "mounted," "connected," "supported," and "coupled" and variations thereof are used broadly and encompass both direct and indirect mountings, connections, supports, and couplings. Further, "connected" and "coupled" are not restricted to physical or mechanical connections or couplings.

The following discussion is presented to enable a person skilled in the art to make and use embodiments of the invention. Various modifications to the illustrated embodiments will be readily apparent to those skilled in the art, and the generic principles herein may be applied to other embodiments and applications without departing from embodiments of the invention. Thus, embodiments of the invention are not limited to the embodiments shown, but are to be accorded the widest scope consistent with the principles and features disclosed herein. The following detailed description is to be read with reference to the drawings, in which like elements in different drawings have like reference numerals. The drawings, which are not necessarily to scale, depict selected embodiments and are not intended to limit the scope of embodiments of the invention. Those skilled in the art will recognize that the examples provided herein have many useful alternatives that fall within the scope of the embodiments of the invention.

Some embodiments of the invention include systems capable of delivering all of the micronutrients in microalgae to soil. In some embodiments, microalgae containing water (effluent) may be inoculated into soil so that the micronutrients are immediately bioavailable by the crop plants growing in the soil. In some embodiments, the system may be placed within an irrigation system between a water source and a water port through which irrigation water may be applied to crops. In some embodiments, the system can produce biofertilizer that can be immediately bioavailable by crops such that negligible runoff contamination occurs. By using the system, inorganic agricultural chemicals can be used more efficiently after being converted into a bioavailable form by algae; thus, the amount of chemicals required is reduced.

In some embodiments, the system can be used to construct soil organics with nutrient-rich algal biomass to restore lean (nutrient-lean) soil. In some embodiments, the system may facilitate and accelerate the transition from chemical-based farms to organic farms. In some embodiments, the system can deliver soil carbonate-solubilised microalgae to the soil, establish polysaccharide content in the topsoil, and increase soil porosity to 500% or more. In some embodiments, the system also provides for the use of specific algal biotoxins in place of conventional chemical fungicides and other chemical poisons/toxins to manage nematodes and other harmful pests.

Some embodiments include systems that may include one or more bioreactors. In some embodiments, the system may comprise a plurality of bioreactors. In some embodiments, when there are multiple bioreactors, the bioreactors may be the same or different. Also, in some embodiments, the contents of the bioreactors may be the same or different. In some embodiments, the culture medium in the bioreactor of the system can comprise one or more types of microalgae. Some embodiments of the invention include embodiments in which: a) all microalgae are of the same type; b) there are two or more different types of microalgae; and/or c) one or more bioreactors comprise one or more types of microalgae and one or more other bioreactors comprise one or more other types of microalgae.

In some embodiments, the microalgae in the bioreactor may be propagated, and thus the initial microalgae inoculant placed in the bioreactor may provide an unlimited supply of microalgae. In this case, the microalgae feed and water may be loaded into the bioreactor, and a sufficient amount of microalgae biomass may be periodically removed from the bioreactor to maintain conditions within the bioreactor suitable for microalgae cultivation.

In some embodiments, the systems and methods of use thereof can increase overall crop yield by 5% to 30% or more compared to untreated crops. In some embodiments, the systems and methods used can improve the texture, taste, size, nutrients, and/or yield of a crop compared to an untreated crop. In terms of agricultural use, in some embodiments, the systems and methods of use thereof may reduce overall energy consumption, and/or reduce ecological pollution, and/or reduce greenhouse gas emissions, and/or increase the bioavailability of micronutrients and macronutrients, and/or reduce the use of fertilizers, and/or reduce overall crop production costs, and/or reduce farming costs, and/or reduce the need for and use of fungicides, herbicides, and/or pesticides, and/or reduce soil compaction, and/or improve soil porosity, and/or increase the microbial content of soil, and/or increase the organic content of soil, and/or reduce the amount of irrigation water required to grow crops, and/or reduce the occurrence of over-fertilization, and/or reduce runoff and soil erosion, and/or improve plant characteristics and/or improve water/moisture retention of the soil, all as compared to untreated crops and farmlands.

In some embodiments, the system can be used to reduce or eliminate carbonate accumulation in irrigation equipment by flowing microalgae-containing water through the irrigation equipment. In some embodiments, the system can also be used to reduce or eliminate the accumulation of carbonates in soil by inoculating the soil with water containing microalgae.

Depending on the crop requirements, microalgae of many different species and strains can be used. Algae can be collected and cultured from fields or commercial sources where crops are grown. Microalgae samples can be obtained from repositories of Arizona State University, University of California at Berkeley, University of Texas at Austin, Wood House academic Research Institute, Scripps Institute of Oceanogray, or other repositories.

Microalgae of different species and strains grow best under different conditions. The culture conditions within the bioreactor will vary depending on the particular species of microalgae present in the bioreactor. The conditions for Culturing many different types of Microalgae can be found in The Handbook of Microalgae Culture: Biotechnology and Applied Phytology (compiled by Amos Richmond, Blackwell Publishing, Oxford, U.K.,2004), Algal Culture Techniques: A Book for All Phytology (compiled by Robert A. Andersen, Elsevier Academic Press,2005) and Microalgae: Biotechnology and Microbiology Cambridge Studies in Biotechnology (compiled by E.W.Becker. Press Synthesis of The University of Cambridge,1994), The disclosures of which references are hereby incorporated by reference in their entirety.

In some embodiments, the native microalgae species may have characteristics that optimize their growth under the environmental conditions of the target geographic location. In some embodiments, algae from a non-local site or collection of algae may be used to inoculate soil at a target geographic location to maximize specific bioavailable compounds. Some embodiments include a method of inoculating soil, which may include: obtaining a soil sample from the target geographical location, and/or isolating robust native microalgae species from the sample, and/or culturing the microalgae to form the first inoculum. Additionally, the method may include inoculating the portable microalgae-based soil inoculation system with the first inoculum and/or culturing the microalgae in the inoculation system to form a second inoculum and/or inoculating soil of the target geographic location with the second inoculum one or more times. Further details are disclosed below.

In some embodiments, the systems of the present invention may employ a variety of different types of water as a source of water, including, but not limited to, wastewater, and/or well water, and/or lake water, and/or brook water, and/or pond water, and/or storm water, and/or river water and/or fresh water. Since water is intended for crop growth, it is preferred that the water source has low salinity and is free of heavy metals. In some embodiments, after leaving the microalgae inoculation system, the inoculum-containing water may be delivered to the crop by any conventional irrigation means or system used in agriculture, for example by an irrigation system of the overflow, sprinkler or drip irrigation type or by a sprayer or aerial application. If applied by spray or aerial application, the treatment may be followed by sufficient water to drive the algae into the soil.

In some embodiments, the systems and methods can provide continuous, semi-continuous, repeated, or periodic treatment of soil with an inoculum containing microalgae. For example, in some embodiments, the soil may be treated with the microalgae-containing inoculum daily, or every other day, or every third day, or every half week, or every fourth day, or every fifth day, or every sixth day, or every week, or every two weeks, or every three weeks, or every four weeks, or every month, or every two months, or every three months, or every half year, or every year. In some embodiments, the soil may be treated with water that does not contain microalgae and then inoculated with water containing microalgae, or vice versa. Some embodiments include diluted, semi-concentrated, and concentrated algal cultures with a single algal species or two or more different algal species. In some embodiments, although optional, the irrigation water may comprise additional crop nutrients (macronutrients and/or micronutrients) in addition to the microalgae feed. For example, in some embodiments, nutrients such as calcium may be incorporated into the algae species for importation and uptake by crops. The following table includes examples of macronutrients and micronutrients.

Algae are symbiotic with other organisms, including microorganisms and macroorganisms. Although the main objective of the present invention is to grow algae, growing algae in different communities of various microorganisms can provide a useful solution. Azotobacteria (known as azotrophs) fall into two broad categories, free-living and symbiotic. Aerobic nitrogen-fixing bacteria have over 50 genera, including Azotobacter (Azotobacter), methane-oxidizing bacteria, and cyanobacteria (cyanobacteria), which require oxygen to grow and, when present, fix nitrogen in the soil. Azotobacteria, some related bacteria and some cyanobacteria fix nitrogen in ordinary air, but most members of this group fix nitrogen only when the oxygen concentration is low. In algal cultures, Aphanizomenon flos-aquae (Aphanizonen flowers-aquae) reduces acetylene and fixes nitrogen. Some symbiotic bacteria belong to the genus Rhizobium (Rhizobium) such as the species Bradyrhizobium (Bradyrhizobium) and Sinorhizobium (Sinorhizobium) which colonize the roots of legumes and stimulate the formation of nodules where they fix nitrogen in a microaerophilic manner. Green microalgae provide nitrogen, phosphorus, potassium, calcium, and various other micronutrients. Thus, some embodiments include embodiments wherein one or more microalgae are co-cultured with one or more nitrogen-fixing bacteria (diazotrophs), or inoculated into the soil with one or more nitrogen-fixing bacteria.

In some embodiments, suitable microorganisms that may be co-cultured with microalgae and/or algae or inoculated into the soil may include actinomycetes, bacteria, fungi, and/or mycorrhiza (mycorrhizae). For example, some embodiments include actinomycetes, which are filamentous bacteria that look like fungi. Although not as numerous as bacteria, they play a vital role in the soil where they help break down organic matter into humus which slowly releases nutrients. They also produce antibiotics against root diseases. The same antibiotics can be used to treat human diseases. When the soil is cultivated, actinomycetes produce the sweet soil odor of bioactive soil.

Some embodiments may include the use of bacteria that break down complex molecules and enable plants to absorb nutrients. Some species release N, S, P and trace elements from organic matter. Other species break down soil minerals and release K, P, Mg, Ca and Fe. Other species produce and release natural plant growth hormones, stimulating root growth. A few bacteria fix N at the roots of leguminous plants, while others fix nitrogen independently of the flora. The bacteria are responsible for converting N from ammonium to nitrate and then back again depending on soil conditions. The various bacterial species increase the solubility of nutrients, improve soil structure, combat root diseases, and detoxify the soil. In some embodiments, bacteria suitable for co-cultivation with microalgae and for use in the present system are disclosed in U.S. patent No. 7,736,508 (6/15/2010) to Limcaco, the relevant disclosure of which is hereby incorporated by reference.

Some embodiments may include the use of fungi, some of which may appear as linear colonies, while others are single-celled yeasts. Slime molds and mushrooms are also fungi. Many fungi aid plants by breaking down organic matter or releasing nutrients from soil minerals. Fungi often colonize large organic masses very early and begin the decomposition process. Some fungi produce plant hormones, while others produce antibiotics including penicillin. Several fungal species trap harmful plant parasitic nematodes.

Some embodiments may include the use of mycorrhiza, which is a group of fungi that live on or in the roots of plants and act to extend root hairs into the soil. Mycorrhiza increases water and nutrient absorption, especially in less fertile soils. Roots colonized by mycorrhiza are less likely to be penetrated by root-feeding nematodes because such pests cannot penetrate thick fungal networks. Mycorrhiza also produces hormones and antibiotics, promotes root growth and inhibits disease. Fungi benefit from flora by absorbing nutrients and carbohydrates from the roots of the plants they live on.

In addition to soil restoration or nutrient supplementation, some embodiments of the systems and methods may also be used to replace or reduce the need for conventional herbicides, insecticides, fungicides, and nematicides. For example, in some embodiments, algae species with specifically selected toxins can be used to manage nematodes and other soil predators after harvesting. Algae with toxins are naturally occurring and usually die after killing nematodes. Although the algae may mutate, the native algae are much more robust and quickly dump any remaining toxic algae. Microalgae suitable for use as pesticides include algae of the genera Nostoc (nosoc), pseudocladia (Scytonema) and leptospirillum (Hapalosiphon). Some embodiments may include the use of the systems and methods in places such as soil-based farms, parks, hydroponic farms, aquafarms, nurseries, golf courses, sports fields, orchards, gardens, zoos, and other such growing crops or plants. Some embodiments described by Duke et al ("Chemicals from Nature for Weed Management", Weed Science, (2002) Vol.50, p.138-151) may include the use of additional phytotoxins obtainable from microorganisms. Some non-limiting examples of phytotoxins include actinonin, brefeldin, carbocyclicins, cerulenin, coelicopin, coelicoquinone (cochlioquinone), coronatine, 1, 4-cineole, fischerellin, fumonisin (fumosin), cystatin (Fusicoccin), garabulin, gotatin (gostatin), megacolinol, hydnocidin, cellosolve, phaseolin, glufosinate, podophyllotoxin, preelmendohorse, pyrazoxazine (pyridazocidin), quassin, rhizobium toxin, marigolds toxin, sorgholone syringoxin (sorgholone sygotoxin), tenutoxin, tricolorin a, thiolactamycin, and usnic acid.

Some embodiments may include the use of bioreactors adapted to receive and use natural and/or artificial light. Thus, in some embodiments, the bioreactor may be adapted to allow exposure of the microalgae to a light source. In some embodiments, the walls of the bioreactor may comprise a light transmissive material to allow exposure of the microalgae to light. If an artificial light source is used, the light source may be placed inside or outside the bioreactor, for example according to U.S. patent No. 8,033,047, the entire disclosure of which is hereby incorporated by reference. Alternatively, in some embodiments, the system may include a water conduit through which the microalgae-containing water in the bioreactor may be circulated to expose the microalgae to light. Some embodiments may include the use of water conduits adapted to use sunlight, reflected light, bent light, fiber optic light, or artificial light.

In some embodiments, the system may be operated continuously, semi-continuously, or batch-wise.

In some embodiments, the system may further comprise one or more monitors or sensors adapted to monitor: a) growth conditions within the bioreactor; and/or b) microalgae cell titer/cell count in water; and/or c) the pH of the water; and/or d) salinity of the water; and/or e) the presence of unwanted microorganisms in the bioreactor; and/or f) water level; and/or g) water pressure; and/or h) the level of microalgae nutrients; and/or I) solids level in the filtered water; and/or j) the level of unwanted compounds in the water; and/or k) oxygen, ozone and/or CO in water₂Content (c); and/or l) the level of nitrogen compounds in the water; and/or m) the transparency or opacity of water; and/or n) the level of one or more desired compounds in the water; and/or o) water flow rate; and/or p) weed algae (weed algae); and/or q) algae predators; and/or) other contaminants.

In some embodiments of the present invention, the substrate is,a monitor or sensor may be used to regulate the operation of the control system, such as by feedback. In some embodiments, the monitor may generate one or more signals to a controller that controls the flow of material into and/or out of the system. For example, in some embodiments, the microalgae cell titer monitor can send one or more signals to one or more flow controllers that control the flow of source water or microalgae-containing water into and/or out of the system. In some embodiments, the pH monitor may be directed to CO₂The flow controller sends one or more signals that the controller controls the CO to be delivered₂Amount or rate of addition to the system. In some further embodiments, the water level monitor may send one or more signals to a water flow controller that controls the amount or flow rate of water into and/or out of the system. In some embodiments, the pH monitor may send one or more signals to an acid or base titration unit that controls the amount or rate of flow of acid or base into and/or out of the system.

In some embodiments, the water pressure monitor may send one or more signals to a water pressure regulator that controls the amount or flow rate of water into and/or out of the system. In some embodiments, the ozone monitor can send one or more signals to an ozone flow controller that controls the amount or rate of ozone added to the system. In some further embodiments, the transparency monitor may send one or more signals to a water transparency controller that controls the filtration efficiency of water in the system. In some other embodiments, the nutrient monitor can send one or more signals to a nutrient source flow controller that controls the amount or rate at which the microalgae nutrient is added to the system.

For growth, plants and microalgae require nutrients such as oxygen, carbon, nitrogen, phosphorus, potassium, magnesium, sulfur, boron, copper, chloride, iron, silicon, sodium, manganese, molybdenum, zinc, cobalt, vanadium, bismuth, iodine, water, carbon dioxide, air, and/or other substances.

The profile of macronutrients and micronutrients provided by microalgae will depend on the strain or species of microalgae used. Plants may require a different spectrum of micronutrients and macronutrients at different stages of the plant life cycle. Some embodiments provide methods of growing crops in which the macronutrient and micronutrient profiles of microalgae are matched to specific stages in the plant life cycle. In some embodiments, the field may receive a conventional nutrient feed during crop growth and development, using different species depending on the needs of the crop. For example, microalgae a provides a nutrient profile a, microalgae B provides a nutrient profile B, and the target crop requires nutrient profile a early in growth and nutrient profile B loop late in growth. In this case, the soil in which the crop is grown will be inoculated with microalgae a first at an early stage of growth of the target crop and then with microalgae B at a later stage of growth of the target crop.

Some embodiments include a method of producing a crop, comprising: planting a crop into soil and inoculating the soil with a first microalgae that provides a first nutrient profile; and/or allowing the plant to enter the second growth stage from the first growth stage; and/or inoculating the soil with a second microalgae that provides a different second nutrient profile. In some embodiments, the first nutritional profile is optimal for plant growth during the first phase and the second nutritional profile is optimal for plant growth during the second phase.

Fig. 1 depicts a first embodiment of a portable microalgae-based soil inoculation system 1 of the present invention. In some embodiments, the system includes a water source 7, an ozone source 2, a carbon filter/ultraviolet light system 3, a water pump 8, a solids filter 9, a

microalgae nutrient source

4a, 4b, a

bioreactor

6a, 6b, 6c, a carbon dioxide source 5, a pressurized air supply/air pump 10, and various water conduits. In some embodiments, the pressurized air supply may be a blower, and/or an air compressor, and/or a rocker pump, and/or any other conventional producer or source of pressurized air. In some embodiments, air is taken from the atmosphere or tank via inlet 11, inlet 11 optionally including an air filter. In some embodiments, air is passed through the air pump 10 to the ozone source 2, thereby forming ozone-treated air and directing it into the water source 7 to form ozone-treated water. In some embodiments, a carbon dioxide source 5 is also injected into the air to form carbon dioxide treated air, which is directed into bioreactors 6a-6c or into the water entering the bioreactors. In some embodiments, the ozone-treated water is filtered by a solids filter 9, a carbon filter, and/or an ultraviolet light system 3 to form filtered water, the microalgae feed is added to the filtered water by

microalgae feed sources

4a, 4b to form feed water, which is directed to a bioreactor. In some embodiments, during initial start-up,

bioreactors

6a, 6b, 6c are filled with water containing microalgae nutrients and then inoculated with a first inoculum containing microalgae. In some embodiments, carbon dioxide-containing air is injected into the microalgae-containing water in the

bioreactors

6a, 6b, 6 c. In some embodiments, the water in the

bioreactors

6a, 6b, 6c is recycled for a period of time until the microalgae cell titer/cell count reaches a target level suitable for use as an inoculant. In some embodiments, water from system 1 is then flowed into the irrigation water to form microalgae-containing inoculum as an effluent, which is applied to the soil from irrigation system 99.

Various operating parameters may be controlled. For example, in some embodiments, one or more heaters are optionally included in the system to heat water conducted through the system and/or to heat the culture medium in the bioreactor, thereby allowing for culturing of microalgae and use of the system even during cold weather.

In some embodiments, the volume of system water and its flow rate of irrigation water into the irrigation system 99 may be adjusted as needed to provide the appropriate level of inoculation and water penetration into the soil. For example, in some embodiments, a 200 acre field may receive a total daily volume of water of 500 to 1000 gallons at a delivery rate of about 21 to 42 gallons/hour. In some embodiments, the inoculum obtained from the bioreactor (e.g., such as one or more of

bioreactors

6a, 6b, 6 c) may be applied to the soil with or without further dilution. For example, in some embodiments, the operating system 1 may be operated such that all water used for irrigation flows through the bioreactor. Additionally, in some embodiments, the system 1 may be operated such that the inoculum, i.e., the effluent of the

bioreactors

6a, 6b, 6c, is diluted with additional irrigation water prior to application to the soil.

In some embodiments, microalgal cell titers (cell counts) in the bioreactor fluctuate over time; thus, the cell titer of the effluent also varied. Titer provides an important measure of the health and productivity of the unit. Typically, the titer in the effluent may be at least 1,000,000 cells/ml up to 30,000,000 cells/ml. Titers are also species specific and may be above or below the above ranges.

In some embodiments, ozone may be used to destroy unwanted microorganisms present in the irrigation water prior to entering the bioreactor. Any organic contaminants present in the system can be removed by ozonolysis, as described in U.S. patent No. 5,947,057 and U.S. patent No. 5,732,654 to Perez et al. Organic contaminants include herbicides, insecticides, fungicides, and the like. In some embodiments, the ozone source can be an ozone generator. Ozone generators may include Pacific Ozone model 01, Absolute Ozone model Nano, and OZ8PC20 by Ozotech. In some embodiments, the water is treated with ozone as needed, depending on the quality of the water entering the system. In some embodiments, the concentration of ozone in the water will vary with the quality of the water, but with an ozone level sufficient to disinfect the water, prior to filtration through the carbon filter. In some embodiments, treatment of water with ozone can be improved by employing a mixer that mixes the water and ozone.

In some embodiments, a carbon filter and ultraviolet light system are used to remove ozone from irrigation water prior to entering the bioreactor. In some embodiments, carbon filters typically use a minimum of 0.75ft³The activated carbon of (1). In some embodiments, the carbon filter and the ultraviolet light system are flow-through systems. In some embodiments, suitable carbon filters include 0.75ft from Affordable Water (www.affordablewater.us)³"Upflow Carbon Filter System". The ultraviolet systems may include the "CSL series" of Aquafine and the "UVS 3XX series" of UV Sciences (www.aquaneuv.com; Valencia, Calif.). In some embodiments, the uv light system may be used to disinfect water prior to entering the bioreactor and/or destroy ozone, chlorine, or chloramines prior to entering the bioreactor. In some embodiments, the ultraviolet light system can be sterilized by inactivating or killing microorganisms in the water.

In some embodiments, when present, the solids filter may be used to remove solids from irrigation water prior to entering the bioreactor. In some embodiments, the solids filter may be a flow-through filter. In some embodiments, suitable solids and filters may include "X100" bag filters from www.filterbag.com or fromwww.filterbag.comThe "FV 1" bag filter.

In some embodiments, suitable carbon filters and/or solids filters may include, but are not limited to, media filters, disc filters, mesh filters, microporous ceramic filters, carbon block resin filters, membrane filters, ion exchange filters, microporous media filters, reverse osmosis filters, slow sand filter beds, fast sand filter beds, cloth filters, and/or any other conventional filters.

In some embodiments, carbon dioxide may be used as a carbon source for microalgae. In some embodiments, carbon dioxide may be added directly or indirectly to the bioreactor. In some embodiments, the carbon dioxide source may be a tank containing carbon dioxide, a carbon dioxide generator, a carbon dioxide sequestration plant that sequesters carbon dioxide from the atmosphere, or a combination thereof. Alternatively, carbon dioxide captured from air may be used, for example, U.S. patent No. 8,083,836, the entire disclosure of which is hereby incorporated by reference. In other embodiments, the carbon dioxide may be derived from acetic acid and/or calcium carbonate.

The atmosphere contains about 0.035 to 0.04 weight percent carbon dioxide. Although atmospheric air may serve as a carbon dioxide source for the microalgae, the concentration of carbon dioxide is generally too low to sustain rapid proliferation of the microalgae in the bioreactor. Thus, in some embodiments, carbon dioxide may be added to the air fed into the culture medium. In some embodiments, the concentration of carbon dioxide in the air added to the culture medium may generally be in the range of about 1 to 3 wt.%, 1.5 to 2.5 wt.%, 1.8 to 2.2 wt.%, or about 2 wt.%.

In some embodiments, a water pump may be included in the system. In some embodiments, when present, the water pump may facilitate the flow of water through the water conduit and/or bioreactor of the system. In some embodiments, the pressure of the incoming irrigation water is sufficient to drive the water through the system if a water pump is not included.

In some embodiments, an air pump or blower (the terms are used interchangeably herein) may be included in the system. In some embodiments, the air pump may facilitate the flow of air, which may or may not contain carbon dioxide or ozone, through the air conduit, water source, and/or bioreactor of the system.

The size or operating capacity of each piece of equipment making up the system may vary as desired. For example, in some embodiments, a portable system containing a total bioreactor capacity of 500 gallons of culture medium can support 200 acres of land and typically requires a minimum operating capacity of the components shown below: a) ozone source-1.5 g/hr; (dry air); b) solids filter-maximum flow rate-40 g/min, minimum surface area 2ft²(ii) a c) Carbon Filter-minimum 0.75ft³(ii) a d) Pumping water-minimum 10 gal/min; e) air pressurized air supply/air pump-at 60 "H₂Minimum 25cfm at 0; f) microalgae feed source-minimum 1.0x10⁶Individual cells/ml; g) liquid carbon dioxide source-801/week.

Fig. 2 depicts another embodiment comprising a portable system 51, wherein the components of the system 51 are mounted on a trailer. In some embodiments, system 51 includes a water tank 52, a plurality of bioreactors 53, an ozone generator 54, a clarifier 55, a combination filter/uvLight system 56, nutrient feed supply 57, CO₂A source 58, a pressurized air supply 59, and a trailer 60. As shown, a water tank 52, a plurality of bioreactors 53, an ozone generator 54, a clarifier 55, a combined filter/ultraviolet light system 56, a nutrient feed supply 57, CO₂Either of the source 58 and the pressurized air supply 59 may be mounted on a trailer 60.

In some embodiments, the system 51 can accommodate a flow capacity of about 0.35-0.7gal/min and can be used to support fields in the range of 200-1000 acres. In some embodiments, the water tank 52 may receive water from an on-site water source at a farm. In some embodiments, system 51 may include eight bioreactors (500 gallons total capacity), water tanks, air filters, solids filters, carbon filters, ultraviolet light systems, ozone sources, carbon dioxide sources, microalgae nutrient sources, pressurized air supply sources, and water pumps (not shown). In some embodiments, bioreactor 53 may have light-transmissive walls, such that sunlight is used as a light source. In some embodiments, carbon dioxide and air may be bubbled into the lower portion of bioreactor 53, so that the bubbles agitate the media as it rises. In some embodiments, system 51 optionally includes a mechanical agitator. In some embodiments, assuming a water flow rate of about 0.35gal/min, system 51 may provide a minimum of about 800,000 microalgae cells per second by the effluent.

Fig. 3 depicts a side view of another system 65 of the present invention comprising an elevated portable platform 66, a water tank 67, a pressurized air supply 68, an ozone source 69, a clarifier 70, a water filter 71, a nutrient source 72, a carbon dioxide source 73, and a bioreactor 74. In some embodiments, one or more components may be mounted on a platform, and one or more components may be placed on the ground or on one or more other platforms.

Although fig. 2 and 3 depict the

water tanks

52, 67 as water supplies, in other embodiments, a source of flowing water may be used instead; thus, in some embodiments, the system of the present invention optionally includes one or more water tanks as a water supply, or no water tanks as a water supply. Although not depicted in fig. 2 and 3, in some embodiments, the effluent of one or more bioreactors may be fed into the water stream of the irrigation system. In some embodiments, the systems described herein may be placed within a partial or complete enclosure, even if the system is portable.

In some embodiments, the performance of the system of fig. 2 was evaluated in a crop study in which melon crops were planted on 200 acres of land. The land is divided into a control zone and a sample zone (see, e.g., fig. 5A-5B). The control group received irrigation water only and was not treated with microalgae supplements. The sample zone receives only irrigation water containing microalgae supplements. Melon seeds were planted prior to irrigation with algae supplement in the soil. Control plants were irrigated approximately every four days, depending on heat. The sample plants were irrigated according to the same schedule as the control. The plants and various aspects of fruit growth were evaluated five weeks (as shown in fig. 4A) and nine weeks (as shown in fig. 4B) after planting.

In short, crops grown according to the systems and methods disclosed herein produce larger and firmer plants. For example, compare figure 5A (showing control plants) to figure 5B (showing sample plants). In addition, the larger melons of fig. 6B were compared to the control plants shown in fig. 6A. In addition, the sample plants produced more flowers per vine, with improved fruit texture and taste, increased sugar content, increased nutrient content, improved appearance, and increased vitamin a content. The details and results are described in example 1.

In some embodiments, the system may also include one or more monitoring devices for performing functions including, but not limited to, measuring CO₂Flow rate, CO in culture₂Content, O in culture₂Content, pH, cell density and temperature in the culture, measuring macronutrient content in the culture or effluent, measuring micronutrient content in the culture or effluent or measuring microalgae titer in the culture or effluent.

Fig. 7 depicts an alternative embodiment of the system of the present invention. In some embodiments, system 11 is suitable for low, medium, and high volume irrigation applications. In some embodiments, system 11 includes an optional pump 18 adapted to receive water from a pressurized or non-pressurized water source 11 a. In some embodiments, water received from the water source 11a is ozonated in an ozone contactor 12, the ozone contactor 12 receiving ozone from an ozone generator 27 and being directed to a clarifier/filter 19, the clarifier/filter 19 removing precipitated solids from the water. In some embodiments, after clarification, the water is directed to a carbon filter or ultraviolet light system 13 that removes ozone and passes to a mixer 22, which mixer 22 mixes the water with the algae feed obtained from the algae nutrient supply 14. In some embodiments, the algae/water mixture is mixed using air bubbles generated by a pressurized air supply 30, the pressurized air supply 30 directing air to an air diffuser at the bottom of the bioreactor 16. In some embodiments, water containing nutrients is introduced into bioreactor 16 where the microalgae are cultured. In some embodiments, the microalgae-containing effluent exits the bioreactor 16 and passes through a valve 26 that regulates the water flow ratio between the bypass water source line 28 and the bioreactor effluent. In some embodiments, controller 29 controls valve 26 to achieve a desired volumetric flow ratio between untreated source water (from bypass line 28) and effluent to provide an inoculum containing a desired or target microalgae titer.

In some embodiments, the system 11 may include one or more distinct controllers. For example, in some embodiments, controller 20 may include an optional feedback loop in which water that has been improperly ozonated may be fed back into ozone contactor 12 for proper treatment. In some embodiments, the controller 21 may include an optional feedback loop so that water that is not sufficiently clarified may be fed back into the clarifier 19 for proper clarification. In some embodiments, the controller 23 can provide control of the algae nutrient supply 14 to regulate the amount of feed material charged to the water. In some embodiments, controller 25 may provide control of carbon dioxide source 15 through the use of pH probe 24, which carbon dioxide source 15 feeds carbon dioxide into bioreactor 16 to adjust the concentration of carbon dioxide in the water and ensure that the water has the proper concentration of carbon dioxide. In some embodiments, the algae/water mixture may be mixed by using air bubbles generated by a pressurized air supply 30, the pressurized air supply 30 directing air to an air diffuser at the bottom of the bioreactor.

In some embodiments, the system 11 may include a portable platform (or body or frame, not shown) on which the various components of the system are mounted. In some embodiments, each individual component of the system may be replaced individually. Although the components are shown as single components, each component may exist in multiple numbers independent of other components of the system.

Fig. 8 depicts an alternative embodiment of the system of the present invention. In some embodiments, the illustrated system 41 may be suitable for low, medium, and high volume irrigation applications or flow to distribution tanks 37. In some embodiments, the dispensing tank 37 may be mounted on a trailer for ease of carrying. In some embodiments, system 41 includes an optional pump 18 adapted to receive water from a pressurized or non-pressurized water source 11 a. In some embodiments, water from the water source 11a is ozonated within an ozone contactor 12, the ozone contactor 12 receiving ozone from an ozone generator 17. In some embodiments, ozonated water is directed to a clarifier/filter 19, which clarifier/filter 19 removes precipitated solids from the water. In some embodiments, after clarification, the water is directed to a carbon filter or ultraviolet light system 13, ozone is removed, and passed into a mixer 22, which mixer 22 mixes the water with algal fertilizer/additives obtained from the algal nutrient supply 14. In some embodiments, water containing nutrients may be introduced into bioreactor 16, and microalgae may be cultured in bioreactor 16. In some embodiments, the algae/water mixture may be mixed by using air bubbles generated by a pressurized air supply 30, the pressurized air supply 30 directing air to an air diffuser at the bottom of the bioreactor, as previously described with respect to system 11 of fig. 7. In some embodiments, one or more probes 33 may be placed in the culture to measure key parameters including pH, temperature, cell density, water mixing speed, dissolved gases, and nutrients. In some embodiments, optional telemetry device 34 may send the metrics from the probe (monitoring device or controller) to a computer server for remote monitoring. In some embodiments, an optional microscope with telemetry capabilities may assist in remote culture monitoring. In some embodiments, optional telemetry equipment 34 includes an optional microscope with telemetry capabilities.

As used herein, telemetry device 34 may be any device capable of facilitating communication between the system of the present invention and a communication and/or control center located remotely from the system of the present invention or in a different geographic location than the system of the present invention. In some embodiments, telemetry device 34 may employ any type of wireless communication system, and may employ any frequency of light waves, radio waves, sound waves, infrared waves, hypersonic waves, ultraviolet waves, other such wavelengths/frequencies, and combinations thereof. In some embodiments, telemetry device 34 employs an IP network, such as the internet, a GSM (global system for mobile communications) network, an SMS (short message service) network, other such systems, and combinations thereof.

In some embodiments, flow imaging device 32 may generate images of algae, predators, and contaminants in the culture for Quality Control (QC) purposes, and may send this data to telemetry device 34. In some embodiments, the microalgae-containing effluent may exit the bioreactor and pass through a valve 31 that regulates the flow of the bioreactor effluent. In some embodiments, optional dewatering apparatus 35 can concentrate the algae into a slurry of desired density that can flow to irrigation or portable container 37. In some embodiments, the optional microbial mixer 36 enables the user to blend the end product with beneficial bacteria other than algae, fungi, or other organisms 38 that are symbiotic with the algae.

In some embodiments, system 41 may include one or more distinct controllers. In some embodiments, controller 20 may include an optional feedback loop so that water that has been improperly ozonated may be fed back into ozone contactor 12 for proper treatment. In some embodiments, the controller 21 includes an optional feedback loop so that insufficiently clarified water can be fed back into the clarifier 19 for proper clarification. In some embodiments, the controller 23 provides control of the algae nutrient supply 14 to adjust the feed rate of the feed water. In some embodiments, controller 25, through the use of pH probe 24, may control the carbon dioxide source 15 that fills the bioreactor with carbon dioxide in order to adjust the concentration of carbon dioxide in the water and ensure that the water has the proper carbon dioxide concentration. In some embodiments, the system 11 may include a portable platform (or body or frame, not shown) on which the various components of the system are mounted. In some embodiments, each individual component of the system may be replaced individually. Although the components are shown as single components, each component may exist in multiple numbers independent of other components of the system.

In some embodiments, a system similar to system 41 of fig. 8 may be used to reclaim degenerated or waste soil. In some embodiments, the algae and microorganism mixture produced by the system may be applied to the soil surface by irrigation or spraying to restore essential nutrients. Algae and other microorganisms continue to multiply in the soil as long as the soil has moisture. Algae transport micronutrients, attract other microorganisms, and add organic matter (humus) to the soil. In some embodiments, the method may remediate degraded or waste soil.

In some further embodiments, a system similar to system 41 of FIG. 8 may culture other microorganisms in the same culture or in separate containers for blending before the culture flows into irrigation or portable containers.

Fig. 9 illustrates a soil enrichment system 900 according to some further embodiments of the present invention. Some embodiments include a solids filter 919, a water storage tank 912, a disinfection system 917, and a neutralization system 915. The growth inducing system may include one or more nutrient feeds, such as first and

second nutrient containers

920, 962, to add nutrient to the treated water. The bioreactor system may include one or more bioreactors 916 to facilitate inoculation and growth of microorganisms. The systems and methods may include various additional systems and subsystems, such as one or more nutrient solution containers, refrigerators, light sources, blowers (e.g., at least one pressurized air supply), carbon dioxide sources, pumps, valves, fluid conduits, air conduits, gas conduits, air filters, gas filters, control systems, sensors, air conditioning units, exhaust systems, portable enclosures, and/or external storage tanks.

In some embodiments, one or more pumps 918, such as peristaltic pumps, may push irrigation water from the water source 95 through the fluid conduit 910. The water source 95 supplies water to the soil enrichment system 900. The water flowing from the water source 95 may be referred to as an "irrigation water" water source 95 may include any suitable irrigation water source suitable for irrigating plants. In some embodiments, the water source 95 may be under pressure, such as water from a well or a public facility of a city, town, or municipality. In some embodiments, the water source 95 may be substantially unpressurized. For example, the water source 5 may include a stationary reservoir, regenerated wastewater, well water, lake water, stream water, pond water, rainwater, river water, and/or fresh water.

Some embodiments of the soil enrichment system 900 can include an automated cleaning system 970 controlled by the control system. The automated cleaning system 970 may include a cleaning solution container 968 for holding a cleaning solution and a pump 918 for pumping the cleaning solution from the cleaning solution container 968 into the fluid conduit and/or one or more bioreactors 916. In some embodiments, each of the one or more bioreactors 916 may include a dedicated valve for connecting a fluid conduit to a cleaning solution container 968 for cleaning solution.

In some embodiments, one or more bioreactors 916 may be inoculated with the microbial inoculant by any suitable method, such as manual inoculation through port 935 in bioreactor 916. In some further embodiments, neutralized irrigation water containing nutrient solution may be directed into any one or more bioreactors 916 until it reaches a preselected fill level 940.

In some embodiments, light source 945/950 may be configured to project light onto and/or into each of one or more bioreactors 916. In some embodiments, the light source 945/950 may include LED lights in any suitable configuration to provide light to the microbial culture. For example, in one embodiment, first light source 945 can be located within one or more bioreactors 916. In another embodiment, first light source 945 can cover an outer surface of one or more bioreactors 16. In another embodiment, second light source 950 may be outside and adjacent to the outer surface of one or more bioreactors 916.

In some embodiments, a control system suitable for implementing one or more embodiments of the present invention can include a computer system communicatively connected to PLC system 914. PLC system 934 may be communicatively coupled to one or more sensors 933, and may provide measurements obtained by one or more sensors 933 to a processor and/or database for remote monitoring, remote data access, and/or remote control of soil enrichment system 900. The PLC system 934 may similarly be in communication with the pump 918, the valves, the disinfection system 917, the neutralization system 915, the at least one compressed air supply 930, the lamps 950, and/or any carbon dioxide source, and configured to control the pump 918, the valves, the disinfection system 917, the neutralization system 915, the at least one compressed air supply 930, the lamps 950, and/or any carbon dioxide source.

In some embodiments, a carbon dioxide source 966 can be used to provide a carbon source to the microbial culture. Carbon dioxide may be added directly and/or indirectly to one or more bioreactors. Carbon dioxide source 966 may be a tank containing carbon dioxide gas, a carbon dioxide generator, a carbon dioxide sequestration device for sequestering and temporarily storing atmospheric carbon dioxide, or a combination thereof.

In some embodiments, the microbial culture may be released from the one or more bioreactors 916 through an outlet, flow through one or more fluid conduits, and flow into an external storage tank 937 for storage. In various embodiments, external storage tank 937 can comprise an at least partially transparent material, such as high or low density polyethylene, polycarbonate, acrylic, and/or PVC, to allow natural or artificial light to penetrate external storage tank 937 and enter the microbial culture. In some embodiments, external storage tank 937 may include a sterile aeration system to support the health of the microbial culture. In some embodiments, the external storage tank 937 may include a tapered bottom to ensure that the microbial culture is completely drained when the microbial culture is released onto the target area 955.

In some embodiments, external storage tank 937 can include a cooling system, such as a refrigerator, to cool the microbial culture during storage. The refrigerated external storage tank can be configured to receive the microbial culture and/or microbial slurry, maintain its sterility, and store it at any suitable temperature.

In some embodiments, the dewatering device 964 can be configured to deliver the concentrated microorganism slurry to the target area 955 and/or the external storage tank 937. Dewatering device 964 may concentrate the microbial culture by any suitable method, such as, but not limited to: 1) flocculating and precipitating; 2) flotation and collection; and/or 3) centrifugation. Further details and operational characteristics of soil enrichment system 900 are described in U.S. patent application serial No. 15/647,005, which is incorporated by reference in its entirety.

Some embodiments include methods of isolating, selecting, and using characteristic microorganisms in agricultural production areas using any of the systems described herein. For example, some embodiments of the invention include methods of selecting, collecting, and growing algae for transport to an agricultural production area. In particular, in some embodiments, the methods focus on the collection, isolation and/or propagation of specific microorganisms, primarily algae, for mass transport to the same biological community from which the algae is collected. In some embodiments, the agricultural production area containing the biological community may be a field of farmed land, and/or raised beds, and/or greenhouses, and/or golf courses, and/or degraded land, and/or indoor growing facilities. Some further embodiments include collecting, isolating, and/or propagating, and transporting other characteristic microorganisms in addition to or separate from the algae. For example, some embodiments include collecting, isolating, and/or propagating and delivering the bacterial species. Other embodiments include collecting, isolating and/or propagating and delivering fungal species.

In some embodiments of the invention, algae may be transported by various means, including but not limited to canal irrigation, overflow irrigation and/or drip irrigation, and/or various conventional overhead spray techniques, and/or various conventional hydroponic culture techniques. In some embodiments of the invention, the effect of delivering algae to an agricultural production area may be an increase in soil organic matter, and/or an improvement in soil structure, and/or a reduction in water and fertilizer utilization, and/or an increase in crop yield and product nutritional value, and/or an overall improvement in soil health, and/or a reduction in water and chemical runoff, and/or an increase in carbon dioxide uptake by soil from the air.

Some embodiments of the invention include methods of obtaining soil and/or water samples from an agricultural production area, and/or culturing microorganisms from soil samples, and/or selecting desired species from soil samples, and/or propagating the selected desired species in greater numbers and concentrations, and/or delivering live microorganisms back to the agricultural production area (e.g., dispersing live microorganisms in a solution in a soil area of a farm or a biotope).

The following steps constitute non-limiting embodiments of a method for collecting, selecting and propagating characteristic algae from an agricultural production area (e.g., a farm or other plant propagation facility):

some embodiments include the step of collecting one or more quantities of soil from one or more locations in an agricultural production area. In some embodiments, each amount or total amount of soil collected may be about 100 grams. In some other embodiments, the amount may be less than 100 grams or greater than 100 grams.

Some embodiments include the step of collecting one or more quantities of water from one or more locations in an agricultural production area (e.g., from a surface water source). In some embodiments, each amount or total amount of water collected may be about 50 grams. In some other embodiments, the amount may be less than 50 grams or greater than 50 grams. In some other embodiments, at least some water may be collected from a ground water source, a runoff water source, or spring or well water.

In some embodiments, one or more of the water and/or soil quantities may be refrigerated to 35 ° F to 40 ° F prior to a subsequent treatment site (including but not limited to a laboratory or facility).

In some embodiments, about 10 grams of soil or 10ml of water from each sample may be added to a 100ml culture tank containing 75ml of AF6(Watanabe) medium. In some embodiments, more or less soil and/or water may be added to the culture tank. In some further embodiments, AF6(Watanabe) medium may be used in combination. In some embodiments, soil and/or water may be incubated in the culture tank. In some embodiments, the incubation can be performed overnight under exposure to a light source of 100 to 200 PAR. In some embodiments, the light source may comprise or emit wavelengths of about 450nm to 485nm and/or about 625nm to 740 nm. In some embodiments, the exposure may be about 12 to 24 hours per day.

In some embodiments, a portion of the incubated sample can be propagated in agar-coated petri dishes. For example, in one non-limiting embodiment, a sample can be plated with 10 μ Ι _ of sample onto four 100x15mm petri dishes with AF6 agar, with ring sterilization between each streak to dilute the sample. In some embodiments of the invention, the petri dish may be at least partially sealed (e.g., 75% sealed with tape) and placed upside down for one to two weeks in front of a light source of 100 to 200 PAR. In some embodiments, the light source may comprise or emit wavelengths of about 450nm to 485nm and about 625nm to 740 nm. In some embodiments, the exposure may be about 12 to 24 hours per day.

In some embodiments, when an isolated sterile algal colony has grown to a particular size, the algal colony can be aseptically harvested and placed into a sterile test tube with sterile AF6 media. For example, in some embodiments, when an isolated sterile algal colony has grown to a diameter of about 3mm, the algal colony can be aseptically harvested and placed into a sterile test tube with sterile AF6 media.

Some embodiments may include an incubation time of one to two weeks followed by selection of the tube with the highest biomass. In some embodiments, the incubation can be performed while exposed to a light source of 100 to 200 PAR. In some further embodiments, the light source may comprise wavelengths of about 450nm to 485nm and about 625nm to 740 nm. In some embodiments, the exposure may be about 12 to 24 hours per day. In some further embodiments, the temperature may be in the range of about 70 ° F to 80 ° F.

Some embodiments include sub-culturing each tube into a new tube, followed by placing the contents of the original tube into a sterile 500ml bottle containing AF6 media equipped with a sterile air injection. In some embodiments, the sub-culture tubes may be exposed to a light source of 100 to 200 PAR. In some embodiments, the light source may comprise wavelengths of about 450nm to 485nm and 625nm to 740 nm. In some embodiments, the exposure may be about 12 to 24 hours per day.

Some embodiments include incubating the bottles for 3-5 days and selecting the bottle with the fastest growth rate and highest biomass and identifying with the new strain ID. In some embodiments, the incubation can be performed while exposed to a light source of 100 to 200 PAR. In some embodiments, the light source may comprise wavelengths of about 450nm to 485nm and about 625nm to 740 nm. In some embodiments, the exposure may be about 12 to 24 hours per day. In some embodiments, the temperature may be in the range of about 70 ° F to 80 ° F.

In some embodiments of the invention, the strain ID of the incubated sample may be recorded in the strain ID database along with the date and place of collection and any additional algae characteristics. In addition, in some embodiments, a new tube can be inoculated with each newly identified strain and placed in an algae library.

In some embodiments of the invention, further steps may include manual selection processes to enhance growth rate, maximum density, and other desired characteristics. In some embodiments, the artificial selection process may comprise a strain of algae exposed to preferred culture conditions. In some embodiments, strains of algae with increased growth rates, higher maximum densities, or other desirable characteristics may be selected for future use rather than poor quality strains. In some embodiments, the poor quality algal strain may be subjected to an artificial selection process to further increase the growth rate, maximum density, or other desired characteristics.

In some embodiments of the invention, one or more steps may be performed in a laboratory or facility remote from the agricultural production area. In some embodiments of the invention, one or more steps may be performed in a laboratory or facility near or part of an agricultural production area. In some embodiments, all steps may be performed at the same location. In other embodiments, at least some of the steps may be performed at one location, and one or more other steps may be performed at another location.

In view of the above description and the following examples, those of ordinary skill in the art will be able to practice the claimed invention without undue experimentation. The foregoing will be better understood with reference to the following examples. All references to these examples are for illustrative purposes. The following examples are not to be considered as exhaustive, but merely as a few of the many embodiments contemplated by the present invention.

Example 1

Evaluation of melon growth System

The system is used for planting the Yosmeit variety of the Hami melon. About 200 acres of land were filled with irrigation water containing microalgae. Crops were watered every five days in the afternoon due to high ambient temperature (120 ° F). The microalgae were continuously added to the irrigation water each time it was watered. Algae from the phylum Chlorophyta (Chlorophyta) and Cyanophyta (Cyanophyta) were added to the irrigation water at a combined density of 60 billion cells per minute. Algae were cultured in the media shown in the following table.

Melons were harvested and when comparing melons grown according to the present invention with melons not grown according to the present invention, the following observations were obtained.

Measurement of	Description of the invention
		Productivity of production	Melon yield was increased by 20% by weight.
Size of	The fruit diameter increased by 22%.
		Texture of	The meat quality of the fruit is maintained or improved.
Shelf life	The shelf life is prolonged by 4 days.
		Taste of the product	The taste of the fruit is maintained or improved.
Candy	The sweetness of the fruit is improved by 20 percent.
		Appearance of the product	The appearance and color of the fruit are maintained or improved.
Vitamin A	The content of vitamin is increased by 20%.

For control plants and plants grown with the system of the invention, various sizes of melon plants were measured at 9 weeks after planting. The observed dimensions are reviewed below.

Parameter(s)	Control	Sample (I)	Multiple of increase
				Diameter of trunk	0.129 inch	0.38 inch	2.9
Diameter of the stem	0.05 inch	0.125 inch	2.5
				Mean leaf length	2.5 inches	4 inch	1.6
Maximum leaf length	3.5 inches	7 inch	2.0
				Total radius of plant	37.8 inches	87.12 inches	2.3
Total plant height	5.7 inches	15 inches	2.6
				Width of flower	0.9 inch	2.3 inches	2.6
Diameter of melon	2.3 inches	5.5 inches	2.4

The N inorganic fertilizer needed by the melon field irrigated by the seaweed is reduced by 50 percent, and the P and K are reduced by 40 percent. The micronutrient savings are about 70%. Farmers report that soil porosity and porosity are improved by 5 times, which makes the crop roots deeper. The higher soil porosity also allows symbiotic macro-organisms and micro-organisms to enter the field, such as earthworms. The farmer reported that pesticide use in melon fields needs to be reduced by more than 50% because algae infused crops appear to produce their own biopesticides, deterring intruders such as sand flies from destroying adjacent fields. The fungicide use by the farmer is reduced by 70% because algae can guarantee longer roots, which are more resistant to nematodes and other soil pests. Thus, the system of the present invention provides substantial improvements in the characteristics of plants and fruits grown using the system of the present invention.

Example 2

Crop growth using two different microalgae

Before planting crop seeds in the soil, the soil is repeatedly irrigated with an inoculum containing a first species from the chlorophyta of microalgae until the soil acquires the desired characteristics of increasing organic matter and polysaccharides in the soil to increase water retention. The seeds are planted in the treated soil and irrigated repeatedly with an inoculum containing a different second species from the cyanobacterial phylum of microalgae to inject nitrogen separated from the atmosphere into the soil until the crop is mature. The crop is then harvested in a known manner. At this point, a third species, also from the cyanobacterial phylum, is introduced into the irrigation water and delivered to the soil where it produces biotoxins to kill unwanted pests in the soil. The first species of the chlorophyta of microalgae is used to enhance the fertility and other properties of the soil by increasing organic matter in the soil, which enhances the colonization by other microorganisms and macroorganisms that further enhance the soil by converting nutrients to a form more readily available to crops and by increasing the porosity of the soil. A second species from the cyanobacterial phylum of microalgae is used to add nitrogen to the soil, thereby reducing the amount of nitrogen fertilizer required by the crop. A third species from the cyanobacterial phylum is used to eliminate or reduce pest populations in the soil.

Example 3

System employing co-culture of two different microalgae

A system comprising a co-culture of two different strains of microalgae is prepared by preparing a culture medium in one or more bioreactors and inoculating it with one or more blue-green algae (cyanobacteria or cyanophyta) and one or more green algae (chlorophyta). Both algae can be independently unicellular or colony-type; however, single cell species are preferred. Some chlorophyta include those of the class chlorophyceae, including those of the order chaetoptera (chaetopterides), Chaetophorales (chaetophororales), chlamydiales (Chlamydomonadales), Chlorococcales (Chlorococcales), phaeocystis (chlorocystales), Dunaliella (Dunaliella), microsporoles (Microsporales), coleoptera (oedogoniles), Phaeophilales (phaeophyales), chlorella (sphaerophyllales), Tetrasporales (tetrasporoles), or clionales (volvacles). Some species of green algae include Chlorella fusca, Chlorella zofingiensis, certain species of Chlorella (Chlorella spp.), Chlorella citrinum, Chlorella stigmaphora, Chlorella vulgaris (Chlorella vulgaris), Chlorella pyrenoidosa (Chlorella pyrenoidosa), and the like. Some cyanophyta include the orders Chroococcales, Gloeobaterales, Nostocales (Nostocales), Oscillatoriales (Oscillatoiales), Pseudosarcinales (Pseudoanabaenales), and Synechococcales (Synechococcales). The algae are co-cultured with natural and/or artificial light. The titer of algae in the medium was allowed to increase to a target level of about 1MM to 100MM cells/ml. The medium was drained from the bioreactor and mixed with water for irrigation.

As used herein, unless otherwise specified, the term "about" or "approximately" is used to mean ± 10%, ± 5%, ± 2.5% or ± 1% of the specified value. As used herein, unless otherwise specified, the term "substantially" is used to mean "largely," "at least a majority," greater than 70%, greater than 85%, greater than 90%, greater than 95%, greater than 98%, or greater than 99%.

The foregoing is a detailed description of specific embodiments of the invention. It should be understood that although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims. All of the embodiments disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure.

It will be understood by those skilled in the art that while the invention has been described above in connection with specific embodiments and examples, the invention is not necessarily so limited, and that many other embodiments, examples, uses, modifications and departures from the embodiments, examples and uses are intended to be encompassed by the appended claims. The entire disclosure of each patent and publication cited herein is incorporated by reference as if each such patent or publication were individually incorporated by reference. Various features and advantageous aspects of the invention are set out in the following claims.

Claims

1. A culture system, comprising:

a bioreactor adapted to propagate microalgae in a culture broth using at least one of natural and artificial light in combination with at least one nutrient comprising at least one carbon source, wherein the microalgae are freely suspended in the culture broth and form a part of the culture broth; and

a water conditioning assembly;

an algae nutrient supply coupled to the bioreactor; a first controller configured to control fluid flow between the water conditioning component and the bioreactor, the water conditioning component coupled to the bioreactor as an input of supply water and configured to condition the supply water to a particular purity that enables substantially unimpeded growth of microalgae in the broth to a specified microalgae concentration, and wherein the first controller is configured to control delivery of the algae nutrient supply to the bioreactor; and a carbon dioxide source coupled to the bioreactor, wherein the carbon dioxide is injected into the culture broth as a carbon source.

2. The system of claim 1, further comprising a second controller coupled to the probe, the second controller configured to regulate release of carbon dioxide from the carbon dioxide source to the bioreactor based at least in part on one or more measurements from the probe.

3. The system of claim 2, wherein the probe is a pH probe configured to measure the pH of the culture solution.

4. The system of claim 1, wherein the water conditioning assembly comprises an ozone generator coupled to an ozone contactor, wherein the ozone generator is configured to generate ozone and deliver ozone to at least partially ozonate the supply water.

5. The system of claim 4, further comprising a solids filter located upstream of the ozone contactor.

6. The system of claim 5, further comprising a carbon filter and/or an ultraviolet light system located downstream of the solids filter, wherein at least one of the carbon filter and the ultraviolet light system is configured and arranged to at least partially deodorize and oxidize the ozonated feed water.

7. The system of claim 1, further comprising at least one pressurized air supply coupled to the bioreactor, wherein the at least one pressurized air supply is configured to generate bubbles to at least partially aerate and/or agitate the culture solution.

8. The system of claim 7, wherein the gas bubbles comprise CO₂、N₂And O₂At least one of (a).

9. The system of claim 1, further comprising at least one reservoir or tank providing or coupled to an input for a water supply.

10. The system of claim 1, further comprising a mobile trailer supporting at least the bioreactor, the water conditioning assembly, and the carbon dioxide source.

11. The system of claim 1, wherein the microalgae algal nutrient supply comprises at least one of a fertilizer, a macronutrient, a micronutrient, and at least two different microalgae species; and

wherein the macronutrients are selected from the group consisting of phosphorus, nitrogen, carbon, silicon, calcium salts, magnesium salts, sodium salts, potassium salts, and sulfur; and the one or more micronutrients are selected from the group consisting of manganese, copper, zinc, cobalt, molybdenum, vitamins and trace elements; and

wherein the micronutrient comprises at least one of vitamins and minerals added to the regulated supply water.

12. The system of claim 1, further comprising a telemetry system configured for remotely monitoring and controlling at least one of operation of one or more of the first controller, the second controller, the bioreactor, and at least one component or assembly of the water conditioning assembly.

13. The system of claim 1, wherein the artificial light comprises LED light located within and/or near an outer surface of the bioreactor and exposing the microalgae to at least one of light.

14. The system of claim 1, wherein the carbon dioxide source comprises at least one of a tank containing carbon dioxide liquid or gas, a carbon dioxide generator, and a carbon dioxide barrier that sequesters and temporarily stores atmospheric carbon dioxide.

15. The system of claim 1, wherein the algae supply comprises at least one of a first algae type and a second algae type.

16. The system of claim 1, further comprising a flow imaging device coupled to an output of the bioreactor, the flow imaging device configured to create an image of at least one of algae, predators, and contaminants in a culture broth for quality control monitoring.

17. The system of claim 1, further comprising a microbial mixer configured to blend at least one of algae, bacteria, viruses, and fungi with any broth exiting the bioreactor.

18. A method, comprising:

preparing one or more microorganism-containing samples from at least one location of a current or planned plant growth area; preparing at least one cultured sample by culturing a microorganism from said sample;

selecting at least one target microbial species from the at least one cultured sample; propagating the at least one selected target microbial species to increase the concentration of the at least one target microbial species in the at least one culture sample by:

providing a bioreactor adapted for propagating the at least one selected target species in a culture liquid, the at least one selected target species being freely suspended in and forming part of the culture liquid; coupling a feed source to the bioreactor and a first controller for controlling flow between a water conditioning module and the bioreactor, the water conditioning module coupled as an input for supply water to the bioreactor to condition the supply water to a specified purity that allows for substantially unimpeded growth of the at least one selected target species in the culture broth to a specified concentration, and wherein the first controller controls the supply of the feed source to the bioreactor; and providing a source of carbon dioxide coupled to the bioreactor and regulating the release of carbon dioxide from the source of carbon dioxide to the bioreactor, wherein the carbon dioxide is injected into the culture broth as a carbon source such that the at least one selected microbial target species can be propagated.

19. The method of claim 18, further comprising a second controller coupled to the probe and the bioreactor, wherein the second controller regulates release of carbon dioxide from the carbon dioxide source to the bioreactor.

20. The method of claim 18, further comprising delivering at least a portion of the at least one target microbial species to at least a portion of the at least one site.

21. The method of claim 20, wherein the at least a portion of the at least one target microbial species delivered comprises at least one viable microorganism.

22. The method of claim 21, wherein the at least one viable microorganism is an algal species specific to the delivery site.

23. The method of claim 21, wherein the at least one viable microorganism is a viable species selected to restore a normal soil flora mixture of the field.

24. The method of claim 23, wherein the living species of algae are selected according to their particular desired characteristics for improving soil at a delivery site.

25. The method of claim 18, wherein the water conditioning assembly comprises an ozone generator coupled to an ozone contactor, wherein the ozone generator generates ozone and delivers the ozone to at least partially ozonate the supply water.

26. The method of claim 25, further comprising placing a solids filter upstream of the ozone contactor.

27. The method of claim 26, wherein at least one of a carbon filter and/or an ultraviolet light system is positioned downstream of the solids filter, wherein the at least one of a carbon filter and an ultraviolet light system at least partially deodorizes and oxidizes the ozonated feed water; and at least one pressurized air supply coupled to the bioreactor, wherein the at least one pressurized air supply generates bubbles to at least partially aerate and/or agitate the culture fluid in the bioreactor.

28. A method, comprising: sampling algae from an agricultural site; selecting at least one desired algal species from a population of algal plants for propagation, the at least one desired algal species being present at an initial concentration in the population of algal plants at the agricultural site; propagating the at least one desired algae species in at least one bioreactor; and delivering the at least one desired species to the agricultural site to increase the concentration of the algae species to a concentration greater than the initial concentration.

29. The method of claim 28, wherein the at least one bioreactor is adapted to propagate the at least one desired algae species in a culture broth using at least one of natural and artificial light in combination with at least one nutrient comprising at least one carbon source, wherein at least one desired algae species is freely suspended in the culture broth and forms a portion of the culture broth; and an algae nutrient supply coupled to the at least one bioreactor and a controller for controlling flow between a water conditioning assembly and the at least one bioreactor, the water conditioning assembly being coupled to the at least one bioreactor as an input for supply water and conditioning the supply water to a specified purity that enables the at least one desired algae species in the broth to grow substantially unimpeded to a specified concentration, and the controller controlling the supply of the algae nutrient supply to the at least one bioreactor; and

a carbon dioxide source coupled to the at least one bioreactor, wherein the carbon dioxide is injected into the culture broth as a carbon source.

30. The method of claim 29, further comprising a second controller coupled to a probe, wherein the second controller regulates release of carbon dioxide from the carbon dioxide source to the bioreactor based at least in part on one or more measurements from the probe.

31. The method of claim 29, wherein the water conditioning assembly comprises an ozone generator coupled to an ozone contactor, wherein the ozone generator generates ozone and delivers the ozone to at least partially ozonate the supply water.

32. The method of claim 29, wherein a solids filter is positioned upstream of the ozone contactor, wherein the solids filter removes solids from the supply water.

33. The method of claim 29, wherein at least one of a carbon filter and/or an ultraviolet light system is located downstream of the solids filter, wherein the at least one of the carbon filter and the ultraviolet light system at least partially deodorizes and oxidizes the ozonated feed water.

34. The method of claim 33, wherein at least one pressurized air supply is coupled to the bioreactor, wherein the at least one pressurized air supply generates bubbles to at least partially aerate and/or agitate the culture fluid in the at least one bioreactor.