Utilizing dust for sustainable agriculture

Abstract

Disclosed herein are methods and systems for cultivating plants using dust from indoor environments and space habitats. The dust serves as a root substrate or nutrient source for cultivating plants.

Description

[0001] UTILIZING DUST FOR SUSTAINABLE AGRICULTURE

[0002] CROSS REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to, and the benefit of, U.S. Provisional Patent Application No. 63 / 603,912 filed November 29, 2023, which is incorporated by reference herein in its entirety.

[0004] BACKGROUND

[0005] The future of a permanent human presence in space will require an innovative breakthrough in sustainable space agriculture. The International Space Station has continuously occupied low-Earth-orbit for over 20 years, yet it is still reliant on resupply missions from Earth for all of its food. Edible plants have been successfully grown on the ISS, but these systems require a continuous resupply of bulky materials, such as nutrients and growth substrates from Earth. The cost to launch material into space ranges from approximately $1,200 to $10,000 per pound depending on the rocket system utilized. Space-based agriculture is required for this large and continually growing space economy. Space-based agriculture will benefit future space missions and commercial applications on Earth.

[0006] On Earth, agriculture systems are strained by both climate change and increases in the global population and crop production will need to be doubled by 2050 to meet these food demands. The development of additional space agriculture techniques such as learning to cultivate crops onboard spacecraft can also benefit Earth-based agriculture by fostering innovative technologies that utilize limited resources in a suboptimal environment which is critical to future space exploration and sustainability on Earth.

[0007] Therefore, there is a growing demand for innovative solutions to improve sustainability in agricultural systems, driven by the need to reduce reliance on traditional fertilizers and soil-based substrates. With increasing environmental concerns and the push for resource efficiency, the agricultural sector seeks alternative methods to recycle waste materials into valuable resources for plant production. Current challenges include identifying new, untapped sources of nutrients and substrates that can support plant growth while minimizing environmental impact. The exploration of unconventional materials with nutrient potential represents a critical area of research, addressing the dual need for sustainable practices and enhanced food security. Developing methods to repurpose such materials into viable agricultural inputs would fill a significant gap, contributing to more resilient and eco-friendly farming systems. i SUMMARY

[0008] In certain aspects, disclosed herein is a method of cultivating plants using dust, comprising: collecting dust from closed-system environments; preparing a dust-based medium; and growing plants in the prepared dust-based medium.

[0009] In some embodiments, the dust-based medium comprises root substrate or nutrient solution.

[0010] In some embodiments, the closed-system environments comprise residential, commercial, or spacecraft.

[0011] In some embodiments, the dust comprises biological and chemical elements. In some embodiments, the biological elements comprise beneficial fungi and bacteria for plant growth. In some embodiments, the chemical elements comprise potassium, calcium, magnesium, phosphorous or nitrates.

[0012] In some embodiments, the plants comprise radish, lettuce, collard greens, brussels sprouts, or cauliflower.

[0013] In certain aspects, disclosed herein is a method of cultivating plants using dust as a root substrate, comprising: collecting dust from closed-system environments; analyzing beneficial microorganisms in the dust; cultivating plants in the dust; and measuring nutritional content of the cultivated plants in the dust.

[0014] In some embodiments, the nutritional content comprises measuring antioxidant activity in the cultivated plants.

[0015] In some embodiments, the dust is further combined with inert substrates; wherein the inert substrates is acrillite.

[0016] In certain aspects, disclosed herein is a method of extracting nutrients from dust for plant growth, comprising: collecting dust from closed-system environments; suspending collected dust in water to release soluble nutrients; filtering the suspension of collected dust; obtaining filtered nutrient solution; and using the filtered nutrient solution in a hydroponic system.

[0017] In certain aspects, disclosed herein is a method for dual-use dust recycling in agriculture, comprising: collecting dust from closed-system environments, wherein the closed-system environments comprise residential, commercial, or spacecraft; utilizing collected dust as a physical substrate for plant roots; or extracting nutrients from the collected dust for use in hydroponic nutrient solutions.

[0018] In some embodiments, the nutrients are extracted from the collected dust by suspending collected dust in water and filtering the suspension. In certain aspects, disclosed herein is a controlled environment agriculture (CEA) system, comprising: a dust collection module for gathering Dust from Astronauts and Space Habitats (DASH); a dust processing unit to extract nutrients and remove hazardous elements from DASH; a plant growth module comprising a plant pillow system or an aeroponics system, wherein the plant pillow system serves as a root substrate; wherein the aeroponics system serves as a nutrient source; and a microbial analysis unit to monitor microbial communities within DASH and plant growth.

[0019] In some embodiments, the environmental condition module comprises ideal temperature, relative humidity, carbon dioxide and lighting for plant growth. In some embodiments, the temperature is 24°C-28°C. In some embodiments, the relative humidity is 70%-75%. In some embodiments, the carbon dioxide is 1200ppm. In some embodiments, the lighting is 16-hour photoperiods at 150 mol / m3-200 mol / m3.

[0020] In some embodiments, the plant pillow system consists of collected dust mixed with a ceramic arcillite substrate, a polymer fertilizer, seeds, and a wicking tube.

[0021] In some embodiments, the aeroponics system comprises soluble nutrient solution derived from the DASH.

[0022] In some embodiments, the dust processing unit comprises: a nutrient extraction subsystem to solubilize nutrients, wherein the nutrients comprise potassium, calcium, magnesium, or phosphorus from the DASH; and a heavy metal removal subsystem, wherein the heavy metals comprise lead, mercury, cadmium or chromium.

[0023] In some embodiments, the microbial analysis unit comprises: identify and quantify beneficial and harmful microorganisms in DASH; and monitor changes in plant-associated microbiomes during plant growth.

[0024] In certain aspects, disclosed herein is a method of utilizing DASH in spacecraft agriculture, comprising: collecting DASH from onboard filtration system; processing DASH to extract nutrients; utilizing the processed DASH as a root substrate in a plant pillow system or as a nutrient source in an aeroponics system; cultivating plant in the plant pillow or the aeroponics system; and monitoring plant growth, health, and safety for human consumption.

[0025] In some embodiments, the cultivated plant is Lactuca sativa (red romaine lettuce).

[0026] BRIEF DESCRIPTION OF FIGURES

[0027] The accompanying figures, which are incorporated in and constitute a part of this specification, illustrate several aspects described below. FIG. 1 shows that dust is collected from the ISS HEPA filtration covering. On Earth, the collected dust is used to grow red romaine lettuce. Microbial species, chemical elements, and plant health are analyzed throughout the growth process.

[0028] FIGS. 2 A and 2B show examples of fungal (FIG. 2 A) and bacterial (FIG. 2B) diversity in dust collected from the International Space Station.

[0029] FIGS. 3 A and 3B show results of limited nutrient / salt analysis in dust collected from Earthbased environments (Dannemiller et. al.) (FIG. 3 A) and the International Space Station (FIG. 3B). Both results show dissolved concentrations from approximately 30 mg of dust. Note: Earth dust was sieved to 300 pm while the ISS dust was not.

[0030] FIG. 4 shows an overview of plant growth experiments. Planting method 1 will use DASH as root substrate and nutrient source in a plant pillow at various ratios to standard acrillite substrate with no fertilizer added. Planting method 2 will extract soluble nutrients from DASH which will be used in an aeroponics watering system at various dilutions. Other important environmental conditions in the table will be used for both planting methods.

[0031] FIG. 5 shows utilizing DASH as nutrient source and root substrate for plant production on spacecraft will promote sustainability and self-sufficiency to enable future long-duration missions.

[0032] FIG. 6 shows the timelapse growth of radish plants in dust collected from residential home vacuum bags. Plants grow in hydroponics system without additional nutrients.

[0033] FIG. 7 shows preliminary results of growing plants in dust collected from residential homes. Each plant was grown using hydroponics methods and the photos shown represent approximately 14 days after initial planting.

[0034] FIG. 8 shows an overview of how to collect dust from home vacuum bags and use them to grow plants in a hydroponics system. Microbial, chemical, and plant health metrics are analyzed in both dust as well as plant samples.

[0035] FIG. 9 shows an overview of methods to best utilize dust as a substrate for plant growth. Planting method 1 uses dust as both the substrate and nutrient source. Planting method 2 extracts soluble nutrients from dust for use with a rock wool substrate. Both methods are performed in a hydroponics system.

[0036] DETAILED DESCRIPTION

[0037] General Definitions

[0038] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. By “about” is meant within 10% of the value, e.g., within 9, 8, 8, 7, 6, 5, 4, 3, 2, or 1% of the value. When such a range is expressed, another aspect includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another aspect. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed.

[0039] The term “comprising” and variations thereof as used herein is used synonymously with the term “including” and variations thereof and are open, non-limiting terms. Although the terms “comprising” and “including” have been used herein to describe various embodiments, the terms “consisting essentially of’ and “consisting of’ can be used in place of “comprising” and “including” to provide for more specific embodiments and are also disclosed. Throughout the description and claims of this specification the word “comprise” and other forms of the word, such as “comprising” and “comprises,” means including but not limited to, and is not intended to exclude, for example, other additives, components, integers, or steps.

[0040] As used in the specification and claims, the singular form “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. For example, the term “an agent” includes a plurality of agents, including mixtures thereof.

[0041] As used herein, the terms "may," "optionally," and "may optionally" are used interchangeably and are meant to include cases in which the condition occurs as well as cases in which the condition does not occur. Thus, for example, the statement that a formulation "may include an excipient" is meant to include cases in which the formulation includes an excipient as well as cases in which the formulation does not include an excipient.

[0042] "Inhibit," "inhibiting," and "inhibition" mean to decrease an activity, response, condition, disease, or other biological parameter. This can include but is not limited to the complete ablation of the activity, response, condition, or disease. This may also include, for example, a 10% reduction in the activity, response, condition, or disease as compared to the native or control level. Thus, the reduction can be a 10, 20, 30, 40, 50, 60, 70, 80, 90, 100%, or any amount of reduction in between as compared to native or control levels.

[0043] By “reduce,” or “abrogate,” (used interchangeably) or other forms of the word, such as

[0044] “reducing” or “reduction,” or “abrogating” or “abrogation” is meant lowering of an event or characteristic. It is understood that this is typically in relation to some standard or expected value, in other words it is relative, but that it is not always necessary for the standard or relative value to be referred to.

[0045] By “increase” or other forms of the word, such as “increasing,” is meant raising or elevating. It is understood that this is typically in relation to some standard or expected value, in other words it is relative, but that it is not always necessary for the standard or relative value to be referred to.

[0046] As used herein, by a “subject” is meant an individual. Thus, the “subject” can include domesticated animals (e.g., cats, dogs, etc.), livestock (e.g., cattle, horses, pigs, chickens, ducks, geese, sheep, goats, etc.), laboratory animals (e.g., mouse, rabbit, rat, guinea pig, etc.), and birds. “Subject” can also include a mammal, such as a primate or a human. Thus, the subject can be a human or veterinary patient. The term “patient” refers to a subject under the treatment of a clinician, e.g., physician.

[0047] “Control” refers to a sample or standard used for comparison with an experimental sample. In some embodiments, the control is a sample obtained from a healthy subject (or a plurality of healthy subjects), such as a subject or subjects not expected or known to have a particular polymorphism. In additional embodiments, the control is a historical control or standard reference value or range of values (such as a previously tested control sample or plurality of such samples), or group of samples that represent baseline or normal values. A positive control can be an established standard that is indicative of a specific methylated nucleotide. In some embodiments a control nucleic acid is one that lacks a particular methylated nucleotide, and is used in assays for comparison with a test nucleic acid, to determine if the test nucleic acid includes the methylated nucleotide.

[0048] “Detecting” is used herein to identify the existence, presence, or fact of something. General methods of detecting are known to the skilled artisan and may be supplemented with the protocols and reagents disclosed herein. For example, included herein are methods of detecting a nucleic acid molecule in sample. Detection can include a physical readout, such as fluorescence output.

[0049] “Dust” is defined as fine particulate matter generated within built environments, consisting of organic and inorganic substances, microbial communities, and chemical elements. Dust may originate from human activities, environmental materials, and structural components of the environment.

[0050] “Built Environment” is any enclosed or semi-enclosed space occupied or used by humans, including residential homes, commercial buildings, and space habitats, where dust accumulates. “Root substrate” is defined as any medium in which plant roots can grow, providing physical support and facilitating access to nutrients and water.

[0051] Nutrient Source: A material or substance that provides essential chemical elements required for plant growth, including but not limited to nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg).

[0052] “Microbial Composition” is the community of microorganisms, including bacteria and fungi, present within a given material, such as dust or plant tissues.

[0053] “Chemical Composition” is the specific chemical elements and compounds present in a substance, including nutrients beneficial for plants and potential contaminants.

[0054] “Nutrient Solution” is a liquid mixture containing dissolved nutrients extracted from dust, used to support plant growth in hydroponic systems.

[0055] “Hydroponic System” is a method of growing plants without soil, using a nutrient-rich water solution and an inert substrate for root support if necessary.

[0056] “Microbial Safety” is the absence of harmful microorganisms, such as human or plant pathogens, in a product intended for agricultural or human consumption.

[0057] “Toxic Metals” is the elements, such as lead (Pb) or cadmium (Cd), present in dust or plants at levels potentially harmful to human health or plant growth.

[0058] “Phyllosphere” is the above-ground portions of plants, including stems, leaves, and flowers, where microorganisms interact with plant surfaces.

[0059] “Rhizosphere” is the region of soil or substrate immediately surrounding plant roots, where interactions between the plant, microorganisms, and nutrients occur.

[0060] “Metatranscriptomic analysis” is a method of analyzing RNA sequences from a microbial community to study gene expression and metabolic pathways.

[0061] “Nutritional Content” is the concentration and variety of essential nutrients, vitamins, and bioactive compounds in a plant's edible parts, relevant for human health.

[0062] “Antioxidant Activity” is defined as the ability of plant compounds to neutralize free radicals, reducing oxidative stress and providing potential health benefits.

[0063] “Aerobic Plate Count” is defined as the microbiological method to estimate the number of viable microorganisms in a sample under aerobic conditions.

[0064] “Inert Substrate” is a material that provides physical support for plant roots without contributing nutrients, commonly used in hydroponic systems (e.g., perlite or rock wool).

[0065] “Microbial Inoculation” is the process of deliberately introducing beneficial microorganisms into a substrate or nutrient solution to promote plant health and growth. “Space Habitat” is defined as a closed or semi-closed environment designed to sustain human life in space, such as the International Space Station (ISS).

[0066] “Sustainability” is the practice of using resources in a manner that meets present needs without compromising the ability of future generations to meet their own needs, with minimal environmental impact.

[0067] “Phytoremediation” is the use of plants to absorb, stabilize, or detoxify contaminants, such as heavy metals, from their growing environment.

[0068] “Crop Yield” is defined as the total quantity of plant biomass or edible produce harvested from a given area or growth system.

[0069] “Stomata Density” is the number of stomata (pores on the leaf surface) per unit area, an indicator of plant health and gas exchange efficiency.

[0070] “Dilution series” is defined as a series of solutions with systematically reduced concentrations of a solute (e.g., nutrients) to determine optimal concentrations for plant growth.

[0071] “Waste Diversion” is defined as a process of redirecting materials, such as dust, from landfills into productive uses, reducing environmental impact.

[0072] “Pathogen Suppression” is the inhibition or control of disease-causing microorganisms in a substrate, nutrient solution, or plant.

[0073] “Differential Gene Expression” is the comparison of gene activity levels under different conditions, providing insights into biological responses and pathways.

[0074] “Sustainable Agriculture” is defined as farming practices that maintain productivity while conserving resources, minimizing environmental impact, and supporting long-term ecological balance.

[0075] “Remote Communities” are defined as geographic areas with limited access to resources or infrastructure, where alternative agricultural inputs, such as dust, may have significant value.

[0076] “Regenerative Production System” is an agricultural approach that replenishes and restores resources, promoting environmental health and sustainability.

[0077] General Description

[0078] Disclosed herein are methods and compositions for creating sustainable agricultural systems on Earth and in space. These methods and compositions utilize dust collected from indoor environments, as well as outer space (interstellar space) as a root substrate and nutrient source for plant production. In one method, dust can be obtained from Earth-based buildings (closed systems) and can be used to successfully grow plants as a replacement for fertilizers and traditional soilbased substrates for use in sustainable agriculture systems (Example 1). Furthermore, a system of using these methods in agricultural applications can be evaluated using three key components of dust and plants: 1) microbiology, 2) chemical composition, and 3) plant health, nutrition, and safety.

[0079] Also disclosed herein are methods and compositions which utilize dust collected in outer space, such as aboard the International Space Station (ISS) for example, as a root substrate and / or nutrient source for plant growth onboard spacecraft. This can result in a fully self-reliant agriculture system onboard spacecraft which are required for the successful completion of long- duration space missions (Example 2).

[0080] Also disclosed herein is a system comprising a growth chamber or pod that comprises a plurality of sensors and systems for controlling all aspects of plant photosynthesis, such as lighting, atmosphere, nutrient supply, and water supply. This system can comprise the collected dust described herein. In essence, such a system is a plant life-support system that can be used practically anywhere on the planet Earth off the planet. These systems can be used as bioregenerative life-support systems for space exploration and colonization. For example, systems as disclosed herein may be used to grow food to sustain human life during relatively long space journeys, such as multi-year manned missions to other planets, and even after landing on those other planets, to support human life for colonization. This system is also referred to herein as a “controlled environment agriculture” (CEA) system.

[0081] Dust, as defined herein, is a collection of fine, dry particles of solid matter, such as soil, sand, or other inorganic material, or organic material such as tissue fragments. Dust can be any size smaller than about 500 pm. For example, dust can be 10-500 pm, which is considered a “coarse” particle. Coarse particles can therefore be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,

[0082] 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,

[0083] 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,

[0084] 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,

[0085] 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,

[0086] 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137,

[0087] 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,

[0088] 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175,

[0089] 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,

[0090] 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213,

[0091] 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232,

[0092] 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270,

[0093] 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289,

[0094] 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,

[0095] 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327,

[0096] 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346,

[0097] 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365,

[0098] 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384,

[0099] 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403,

[0100] 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422,

[0101] 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441,

[0102] 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460,

[0103] 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479,

[0104] 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498,

[0105] 499, or 500 or more pm, or any amount in between these amounts.

[0106] The dust particle can also be between 1-10 pm, which is considered a ‘fine” particle. In this case, the particle can be 1, 2, 3 ,4, 5, 6, 7, 8, 9, or 10 pm, or any amount below or in-between those values.

[0107] The dust particle can also be less than 1 pm, which is considered a “submicron” particle. In this case, the particle can be 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.5, 0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.6, 0.61, 0.62, 0.63, 0.64, 0.65, 0.66, 0.67, 0.68, 0.69, 0.7, 0.71, 0.72, 0.73, 0.74, 0.75, 0.76, 0.77, 0.78, 0.79, 0.8, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.9, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, 0.99, or 1.0 pm, or any amount in between or below these values.

[0108] On Earth, “dust” generally consists of particles in the atmosphere that come from various sources such as soil lifted by wind, volcanic eruptions, and pollution. In built environments, dust can comprise organic tissue fragments, minerals from outdoor soil and many other materials which may be found in the local environment. Some examples of organic tissue fragments include but are not limited to, small amounts of plant pollen, human hairs, animal fur, textile fibers, paper fibers. Space dust, also known as cosmic dust or extraterrestrial dust, can be made of made of rock, ice, minerals, or organic compounds that exists in outer space or has fallen to Earth.

[0109] Dust particles from space can be made of carbon-rich dust, which is more like soot, or silicate-rich dust, which is more like sand. Comets, asteroids, and meteor fluxes are the main sources of interplanetary dust. Dust from outer space can be referred to herein as Dust from Astronauts and Space Habitats (DASH). “DASH” is used interchangeably herein to mean dust collected from outer space.

[0110] Some examples of a “built” environment include but are not limited to a laboratory, a hospital, a manufacturing plant, an airport, an airplane, a school, an office, a vehicle, an apartment complex, a dormitory, a barrack, a prison, a spacecraft, or a home. By “closed environment” is meant that the built environment comprises a structure which has a filtration or other system for allowing the passage of air into the building. Because the system is closed, dust particles can be trapped and collected. This system can be used in conjunction with an HVAC (Heating, Ventilation, and Air Conditioning) system, for example. Some HVAC systems are compatible with high-efficiency filters like High-Efficiency Particulate Air (HEP A) or Ultra-Low Penetration Air (ULPA) filters. Any system in a building, car, or other structure which allows for the collection of particles can be used with the methods disclosed herein.

[0111] In some embodiments, collected dust is used as a root substrate and nutrient source for plant growth. DASH is generated onboard the ISS, for example, through the everyday activities of the crew consisting of biological and chemical elements shed from the occupants as well as built environment materials. It is shown herein that DASH contains a diverse fungal and bacterial community that can be leveraged by plants in beneficial ways. In some embodiments, fungi can support plant growth by biocontrol of pathogens with antibiotic production or competition as well as strengthen defense mechanisms of the plants themselves. Plant growth also requires nutrients including but not limited to Aluminum (Al), Copper (Cu), Potassium (K), calcium (Ga), Magnesium (Mg), Phosphorous (P), Iron (Fe), Manganese (Mn), Sulfur (S), Sodium (Na), Nitrogen (N), or Zinc (Zn). House dust samples from Earth-based homes have previously been shown to contain at least some of these nutrients.

[0112] The systems disclosed herein can be used to efficiently grow crops using less resources than traditional agricultural methods while also enabling those crops to be grown in many areas that would traditionally not be able to grow crops. For example, plant growth systems disclosed herein may be used within buildings in the inner city, in the desert where there is little rainfall and crops would normally not be able to be grown in the land, in environments that would typically be too hot or too cold to grow particular crops, and / or the like.

[0113] Collected dust can be used for a number of different agricultural methods. For example, collected dust can be used as a growing medium or substrate. This can be used to grow a variety of plants. They can be started from seed, tubers, or cuttings (transplants). The dust can be used for initial germination, or during the growth phase of the plant, or to transplant a mature plant. The collected dust can be used as an additive to already existing growth media. It can be used in conjunction with fertilizer or other materials or substances which promote germination, growth, or fruit / flower production. In some embodiments, the dust can be used from start to finish for germination through maturity.

[0114] In some embodiments, the plant is grown to produce an edible product. Examples include, but are not limited to, fruit (such as, for example, grapefruit, oranges, mandarins, limes, peaches, apricots, plums, nectarines, bananas, mangoes, strawberries, raspberries, blueberries, kiwifruit, passionfruit, watermelons, rockmelons, honeydew melons, apples, pears, tomatoes, and avocados), vegetables (such as, for example, lettuce, spinach, silverbeet, arugula, kale, chard, parsley, chives, cabbage, cauliflower, Brussels sprouts, broccoli, potato, sweet potato, yam, rutabaga, beet, celery, asparagus, onion, garlic, shallot, cauliflower, broccoli, artichoke, pea, lima bean, eggplant, cucumber, sweet com (maize) , tomato, peppers, acorn squash, butternut squash, patty pan squash, spaghetti squash, and Hubbard squash), oilseeds (such as, for example, groundnut, soybean, palm kernel, cotton seed, olive, sunflower seed, rapeseed, sesame seed, linseed, safflower seed), or cereals (such as, for example, maize, rice, wheat, barley, sorghum, millet, oat, rye and triticale). The plant being grown can also be grown to produce oxygen, for aesthetic purposes, or for any other use for which a plant can be purposed.

[0115] In some embodiments, the method comprises collecting dust from the space or terrestrial environment, wherein space is spacecraft. An example of a spacecraft is the International Space Station (ISS). In some embodiments, the space is a low-Earth orbit or space habitats, wherein space habitats are moon and Mars. “Outer space” is defined herein as being above the Karman line, which is located approximately 100 kilometers (62 miles) above sea level. Therefore, “outer space,” as defined herein, is 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 km or more above sea level.

[0116] In a specific embodiment, collected dust, such as DASH, is vacuumed from the HEPA filter coverings that are part of an air ventilation system, which is where most of it is deposited. These vacuum bags are then stored until dust from the HEPA filtration covers are obtained and used for plant production. Some further examples of sample collection devices include but are not limited to a swab, a brush, tubes with lids, a pair of forceps, a vacuum cleaner with a collection bag, a canister, or a zip-top bag. In some embodiments, collected dust can be deposited directly into a container for plant growth after collection. In other embodiments, the filter can be replaced and the filter itself can be used as the growth substrate.

[0117] In some embodiments, dust is collected weekly. In some embodiments, dust is collected every day, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, every 7 days, or more. In some embodiments, dust is collected every week, every 2 weeks, every 3 weeks, every 4 weeks, or more. In some embodiments, dust is collected every month, every 2 months, every 3 months, every 4 months, every 5 months, every 6 months, every 7 months, every 8 months, every 9 months, every 10 months, every 11 months, every 12 months, or more. In some embodiments, dust is collected every year, every 2 years, every 3 years, every 4 years, every 5 years, or more.

[0118] In some embodiments, collected dust is processed to remove contaminants and to enhance its nutrient content. In some embodiments, the processing step includes filtering, sterilizing, and enriching the collected dust with essential nutrients. Some examples of filtration techniques are gravity filtration, vacuum filtration, centrifugal filtration, hot filtration, cold filtration, granular media filtration, and mechanical filtration. Examples of sterilization techniques include but are not limited to, steam sterilization (autoclave), dry heat, ethylene oxide (EtO) gas, hydrogen peroxide gas plasma, radiation (gamma rays, X-rays), liquid chemical sterilant treatment, filtration as described earlier, and vaporized hydrogen peroxide (VHP). Enriching the collected dust with essential nutrients comprises addition of Aluminum (Al), Copper (Cu), Potassium (K), calcium (Ga), Magnesium (Mg), Phosphorous (P), Iron (Fe), Manganese (Mn), Sulfur (S), Sodium (Na), Nitrogen (N), or Zinc (Zn). In some embodiments, the processed dust is treated to create a suitable growth medium for plants. In some embodiments, the treating step comprises adding organic matter, microorganisms, and other additives to optimize the physical and chemical properties of the dust as a growth medium. In some embodiments, the treated dust is utilized as a nutrient source and root substrate for plant production.

[0119] In other embodiments, the dust is combined with other elements to promote plant growth or cultivation. Examples include the addition of nutrients (such as fertilizer or nutrient solution), water, other soil substrate, sphagnum moss, Styrofoam beads, or other material which can enhance plant growth and / or cultivation. This can be an inert substrate such as acrillite.

[0120] In one aspect disclosed herein is a method of utilizing the dust in a spacecraft, comprising, gathering vacuumed dust particles from the spacecraft environment, processing the collected dust, treating the dust with organic matter, microorganisms, and other additives, cultivating plants with the treated dust as a nutrient source and root substrate, as comprised in any of the preceding aspects. Also disclosed herein is a method of utilizing the dust for sustainable agriculture on Earth. In some embodiments, the method comprises collecting vacuumed dust from terrestrial environments. In some embodiments, the terrestrial environments are industrial facilities or urban areas. Some other exemplary terrestrial environments include but are not limited to forests (temperate deciduous, tropical rainforests, coniferous), grasslands, deserts, tundra, taiga (boreal forests), savannas, alpine regions, and even man-made environments like farms and cities. In some embodiments, the dust is collected using a vacuum cleaner with a collection bag. Some further examples of sample collection devices include but are not limited to a swab, a brush, tubes with lids, a pair of forceps, a canister, or a zip-top bag. The method further comprises processing and treating the collected dust to create a nutrient-rich growth medium, utilizing the treated dust as a sustainable alternative to traditional soil for plant and crop cultivation, as described in any of the preceding aspects.

[0121] In some embodiments, the method further comprising the step of monitoring the plant microbiome before and after exposure to the vacuumed dust to assess any changes in microbial communities. Monitoring can include the use of several tools and techniques such as microscopy, rRNA, or rDNA sequencing, fingerprinting, probing, clone libraries, chips, and metagenomics.

[0122] In some embodiments, the nutritional value of plants grown with the addition of vacuumed dust as a nutrient source and root substrate is evaluated. In some embodiments, the method is to assess the concentration of essential nutrients in plants grown with the vacuumed dust. In some embodiments, the essential nutrients are Aluminum (Al), Copper (Cu), Potassium (K), calcium (Ga), Magnesium (Mg), Phosphorous (P), Iron (Fe), Manganese (Mn), Sulfur (S), Sodium (Na), Nitrogen (N), or Zinc (Zn). In some embodiments, the nutritional analyses include determining the levels of macronutrients and micronutrients in plants. This procedure is often performed using spectrochemical techniques such as Flame Atomic Absorption Spectroscopy (FAAS), Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) or Agilent 4200 / 4210 Microwave Plasma Atomic Emission Spectrometer (MP-AES).

[0123] In one aspects, disclosed herein is a method for assessing the safety of plants cultivated with vacuumed dust for human consumption, comprising, determining the presence of any plant or human pathogens introduced through the dust, chemical analyses to identify and quantify any potentially harmful chemicals that may have entered the plants from the vacuumed dust. In some embodiments, the chemicals are Chlorine (Cl), Boron (B), and Molybdenum (Mo), Lead (Pb), Mercury (Hg), cadmium (Cd), Nickel (Ni), Silver (Ag), Tin (Sn), Zirconium (Zr), Gallium (Ga), Chromium (Cr), or Cerium (Ce) as described in any of the preceding aspects. EXAMPLES

[0124] Example 1: Dust Recycling for Environmental Agriculture Materials (DREAM)- using dust collected from indoor environments as a root substrate and nutrient source for plant production.

[0125] House dust samples from Earth-based homes has been shown to contain at least some key nutrients (FIG. 3 A), but no studies on dust have looked for nutrients specific to plant growth. Dust has previously been shown to consist largely of carbonaceous elements, but also significant amounts of metals from the spacecraft built environment have been found. The current study on dust also measured and found some nutrients, but further research is needed to understand this more thoroughly (FIG. 3B). Results shown here show the successful growing of radishes, lettuce, collard greens, brussels sprouts, and cauliflower in dust collected from vacuum bags of residential homes (FIGS. 6 and 7). The primary goal of this study, Dust Recycling for Environmental Agriculture Materials (DREAM), is to further close the sustainability loop of agricultural systems on Earth and in space, by using dust collected from indoor environments as a root substrate and nutrient source for plant production. Dust is obtained from Earth-based buildings to determine its ability to successfully grow plants as a replacement for fertilizers and traditional soil- based substrates for use in sustainable agriculture systems. To accomplish this goal, three key components of dust and plants are evaluated: 1) microbiology, 2) chemical composition, and 3) plant health, nutrition, and safety.

[0126] Establish best methods to utilize dust as a substrate for growth.

[0127] Two techniques are evaluated to use dust as a nutrient source for plant growth. A full microbial profile of dry dust used as a root substrate and nutrient solution extracted from dust is determined before, as well as after, use for plant growth. In addition, plant-associated microbial components are measured in the phylloshphere (roots, stems, and leaves) and pseudo-rhizoshpere (usually soil, but in this case dust). Analyses include quantification and identification of fungi as well as bacteria. A metatranscriptomic analysis of microbial RNA is performed to highlight metabolic pathways including those relevant to plant health, nutrient cycling, and human health. The chemical composition of the dust is analyzed, specifically for nutrients relevant for plant growth. This is done on the original dust samples from the indoor environment and in the nutrient solution extracted from dust.

[0128] Planting methods and screening-This study demonstrated successful germination and growth of radish plants as well as others in the Brassica family. For DREAM to be a successful and viable planting option, comprehensive testing of multiple nutritious plant groups is needed. A variety of vegetable and cereal plants can be tested including, but not limited to, various lettuce varieties, tomato, beans, rice, and wheat. Eight to ten publicly available varieties per plant type that covers both fresh and processed vegetable as well as small grain and with varying stress tolerance (e.g., drought and heat) are tested. Plant cultivation is carried out at Ohio State’s Controlled Environment Agriculture Center at Waterman Farm. Two methods are tested that utilize dust for growing plants. Planting method 1 uses dust as both a nutrient source and a root substrate. Dust is taken directly from a residential home vacuum bag and seeds are planted directly into this for germination. The dust is collected as done previously, which was determined to be “not human subjects research” by The Ohio State University Institutional Review Board. This method utilizes 5 scenarios that include dust only as well as a 1 : 1, 2:1, 3: 1, and 4: 1 mixture of dust / acrillite. No fertilizer is added to any of the scenarios using dust and the specific quantity of dust used are determined based on the density of dust received. A positive control is included that uses the standard substrates, such as rock wool, which is used in hydroponics systems. Planting method 2 extracts soluble nutrients from dust for use as the sole nutrient source (fertilizer replacement) in the hydroponic system while using standard substrates as mentioned above. Dust is submerged in DI water and placed on a shaker table for ~12 hours at which point it is put through a 0.45 pm filter. The collected solution is used as the water / fertilizer in the hydroponics system at a IX, 5X, 10X, 50X, and 100X dilution series. A positive control with a pre-made nutrient solution is also used for comparison for plants grown with dust-based nutrients. All plants are grown in a non-aerated, non-circulated hydroponics system as previously described. A total of 72 plants are grown that includes 6 plants for each scenario from both planting methods.

[0129] Microbial Analyses of dust and plants-DNA is extracted from dust and plants using a Maxwell® RSC PureFood GMO Authentication and Plant DNA kit respectively, with a modified extraction protocol. Dry dust samples and nutrient solutions extracted from dust are measured for all planting methods and scenarios before as well as after cultivation. After plants are harvested microbial analyses on leaf, root, and stem samples are performed. For all planting scenarios using dust, a negative control is used which does not contain plant seeds. Quantification of total fungi and bacteria are completed using qPCR on an Applied Biosystems Quantstudio 6 Flex and analyzed using Quantstudio Real-Time PCR Software vl.3. Total fungal concentration is measured using Aspergillus fumigatus as a standard and universal fungal primer set targeting the ITS region. Total bacterial concentrations are measured using a Bacillus atrophaeus standard and a primer / probe set targeting the 16S rRNA gene. Microbial composition are analyzed using DNA amplicon sequencing for ITS (fungal) and 16S (bacterial) regions as previously described. Processing sequence reads and assigning taxonomy can utilize a well- established bioinformatics pipeline incorporating QIIME2, BLAST, and FHiTINGS. Microbial function is measured by performing a metatranscriptomic analysis on a subset of samples from dust and plants. Specific samples that reflect plants that grew well and those that did not are determined. RNA is extracted using a Maxwell® RSC Plant RNA kit and sent to the Yale Center for Genomic Analysis for sequencing. An Illumina NovaSeq 2x100 lane with 5 million reads per sample is used for sequencing of bacterial, fungal, and plant RNA. The Trinity pipeline with DESeq2 is used to create differentially expressed transcripts and clusters according to gene expression profiles. For these analyses, contigs from CD-HIT-EST are annotated with Trinotate, a suite designed for functional annotation of de novo assembled transcriptomes. Common protein domains are identified using BLAST+ with Swiss-Prot and PF AM databases. Using the SQLite database, a Trinotate annotation report is generated to produce Gene Ontology (GO) and KEGG Ontology (KO) terms. This data is mined for processes related to nutrient cycling, virulence factors, and plant metabolites which can help to understand what is happening during the cultivation of plants.

[0130] Chemical analyses of dust and plants-Chemical composition in dust is evaluated in dry dust samples (Planting Method 1) and extracted nutrient solutions (Planting Method 2). These samples are measured for nutrients relevant for plant growth (N, P, S, etc.) and potential growth inhibitors such as aluminum. Plant stems, leaves, and roots are also measured to determine how much nutrient uptake is occurring from each planting method as part of Objective 2 below. A modified EPA method 3050B is used for nutrient specific element extractions on dry dust and plant leaves. In addition, the edible portion of plants are measured for chemical hazards which may be harmful to human health such as toxic metals. Toxic metals in leaves are extracted using the U.S. Geological Survey digestion procedure. Elemental analyses are performed using a Agilent Technologies 5100 Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). All measurements on ICP-OES utilize a multi-element standard solution containing all elements of interest. A list of all chemical measurements that are made in each sample type are shown in Table 1. This screening can determine the best method to grow plants using dust as a sustainable nutrient source. Following this screening, a microbial and chemical characterization of the growth that can be used is completed, which can help inform future practices using this method. Ultimately, this project can result in a novel and sustainable method to use an overlooked waste product (dust) as a nutrient source to support plant growth, both on Earth and in space. Table 1: Chemical analysis performed during the experiment.

[0131] Evaluate plant health, nutrition, and safety for human consumption.

[0132] The overall health of plants is evaluated using a variety of qualitative observations such as crop yield, plant height, leaf area, and stomata density. Nutritional content is assessed by measuring antioxidant activity and nutritional content in edible plant parts. To determine if plants are safe for human consumption, the microbial content is evaluated following established NASA requirements for ISS non-thermostabilized food requirements.

[0133] This project provides a novel method for sustainable agricultural nutrient management. The need for sustainable agriculture systems is imperative for food production on Earth as well as future space exploration missions to the Moon, Mars, and beyond. These agriculture systems are often limited by nutrient management which is typically provided by using synthetic fertilizers. There are also organic fertilizers that can be used from sources such as composting, wastewater, and manure. This invention provides a novel source that is contained in almost every human occupied built environment and is already collected on a regular basis: dust. This new source provides a sustainable method to provide critical nutrients to plants without increasing the impart of climate change on the world.

[0134] Estimated dust from commercial buildings in the US alone contributes 2 billion kg to landfills each year, and this number does not even include the 105 million residential buildings in the US. This dust collected from built environments can be used as a nutrient source and substrate for plant production, which can divert trash from landfills. DREAM may also provide a readily available and cost-effective nutrient source for plant production in remote or underserved communities where the rising cost of synthetic fertilizers may be a barrier to entry into agriculture. This project contributes significant knowledge to program area “Foundational Knowledge of Agriculture Production Systems (Al 104)” by addressing several key areas. The knowledge addresses critical concerns regarding system feasibility, logistics, expected output, and safety. Importantly, it creates a new growth substrate that is both sustainable and readily available for almost everyone. It addresses microbial community changes in plants and a new novel growing media in a regenerative production system as well as elucidating plant nutrient uptake / utilization in this system.

[0135] Dust is collected from residential homes in Ohio for use in this study. The dust undergoes comprehensive microbial and chemical analyses specifically related to plant production. Best methods for dust utilization as a substrate for growth are determined, using a hydroponics system with physical dust substrate and using only extracted nutrients from dust with bead substrate. Once the best dust utilization is determined, a variety of plants are screened to determine which have the best growth potential with this novel production method. For all plants that are successfully grown, the nutrition and safety are evaluated for human consumption. A general project overview can be seen in FIG. 8.

[0136] Plant Health, Nutrition, and Safety- The overall health of plants is evaluated by measuring plant height, leaf area, stomata density, and crop yield. Leaf area and plant height is measured using photographs and scanned images with Imaged software. Stomata density is measured from an epidermal impression sample of the leaves and counted using a 40X objective on a light microscope. Crop yield is determined by calculating the total fresh weight of plants harvested divided by the growing area for each plant. In addition, to the nutrient analyses on plant stems, leaves, and roots described above, the antioxidant activity of plant leaves are measured. Antioxidant activity is a good indicator of nutritional value of produced lettuce for its natural antiinflammatory benefits to human health. Antioxidant activity is determined by measuring total phenolic content and use of a Oxygen Radical Adsorption Capacity (ORAC) assay. Toxic metals in the plant materials are evaluated as described above. The NASA baseline microbial assessment is used to determine safety for human consumption. This includes measuring total viable microorganisms using an aerobic plate count, total yeast, and total mold assays. In addition, specific human pathogens are directly measured, which include Enterobacteriaceae, E. coli, Salmonella sp., and Aspergillus flavus. All protocols for microorganism enumerations and standards for safety are assessed using the Food and Drug Administration Bacteriological Analytical Manual. This microbial data combined with the heavy metal analyses described above helps determine the safety for human consumption of plants grown with dust.

[0137] Alternative strategies-Additional analysis of plants that are known to uptake less metals from the soil and / or target the use of non-food crops (e.g., cotton) can be targeted. Some plant species are capable of phytoremediation of contaminants such as heavy metals in soils. It has been previously reported that some fungal plant pathogens such as Aspergillus and Alterneria sp. can be found in house dust which may inhibit growth of some plants. In addition, some microbial human pathogens may be present in dust, but once water is introduced these can pose a minimal risk due to competition from non-pathogenic species and difficult mobility to edible portions of plants. The system can also be inoculated with a healthy microbiome from plants grown in soil if necessary.

[0138] Statistical Analyses-Chemical quantities, microbial counts, and plant health metrics are compared for each planting method and scenario within that method (e.g., different dilutions for nutrient solution and ratios for plant pillow) using a Kruskal-Wallis one-way ANOVA test. Microbial beta diversity differences is compared using the Bray-Curtis (for fungi) and UniFrac (for bacteria) distance matrices while alpha diversity is compared using richness as well as Shannon diversity metrics. To analyze statistical significance between microbial taxa for each planting method and scenario, a multiple comparisons test in Statistical Analysis System (SAS) MULTTEST with the false discovery (FDR) which has been described previously is used. For metatranscriptome analyses, differentially expressed transcripts based on the most significant FDR and fold-changes are clustered according to their patterns. Pairwise differential gene expression and co-expressed gene clusters relating to each planting condition and scenario are discovered using Trinity pipeline. Pairwise comparisons are then performed for each planting method and scenario.

[0139] Example 2: Dust Recycling for Environmental Agriculture Materials in Space(DREAMS)- using dust collected from space habitats as a root substrate and nutrient source for plant production.

[0140] Human presence in space requires an innovative breakthrough in sustainable space agriculture. Despite spacecraft such as the International Space Station being continuously occupied in low-Earth-orbit for over 20 years, it is still reliant on resupply missions from Earth for all its food. While edible plants have been successfully grown on the ISS, it requires a continuous resupply of materials, such as nutrients, from Earth. Repurposing waste materials present onboard spacecraft such as dust from astronauts and space habitats will create more sustainable spacecraft and reduce reliance on Earth. This study aims to determine if ISS dust can be used as a root substrate and / or nutrient source for plant growth onboard spacecraft. To accomplish this, ISS dust is returned to Earth where it is used to grow red romaine lettuce (Lactuca sativa) through traditional substrate and aeroponics methods. Relevant microbial, chemical, nutritional, and health data are collected from ISS dust and plants grown from it. The results from this experiment can be the first steps in creating a fully self-reliant agriculture system onboard spacecraft which re required for the successful completion of long-duration space missions beyond in and beyond LEO.

[0141] The International Space Station (ISS) has been continuously occupied by astronauts for over 20 years providing essential knowledge for future long-duration missions beyond low-Earth orbit (LEO) and innovations that have direct Earth-based applications. As the ISS is set to retire in 2030, the National Aeronautics and Space Administration (NASA) has implemented the Commercial LEO Destination (CLD) initiative. The CLD is ushering in a new era of human spaceflight which involves a shift to private industries such as Nanoracks LLC, Northrop Grumman, and Blue Origins constructing and maintaining the next generation of spacecraft in LEO. In addition, NASA is in preparation to return astronauts to lunar orbit and establish a habitat on the lunar surface as part of the Artemis missions. The combination of these efforts provides a significant investment opportunity for new technologies. On Earth, agriculture systems are being strained by increases in the global population and it is estimated that crop production will need to be doubled by 2050 to meet these food demands. In addition, climate change is also contributing to reduced agricultural output. For example, rising temperatures dramatically reduce wheat production, increase virulence of plant pathogens, and cause extreme weather events that destroy farmland. Space-based technologies, such as utilizing remote sensing from satellites, have helped to mitigate some of these issues by monitoring water use, nutrient management, plant health, and crop yields. The development of additional space agriculture techniques such as learning to cultivate crops onboard spacecraft can also benefit Earth-based agriculture by fostering innovative technologies that utilize limited resources in a suboptimal environment which is critical to future space exploration and sustainability on Earth.

[0142] In 1982 the first successful plant, Arabidopsis, flowered and produced seeds onboard the Russian spacecraft Salyut-7. Since then, many types of plants have been grown and the ISS currently utilizes Controlled Environment Agriculture (CEA) systems for plant production onboard. CEA is a technology focused system that regulates environmental controls such as lighting, water supply, air flow, temperature, and nutrient delivery within a plant growth chamber. For example, the Vegetable Production System (Veggie) and the Advanced Plant Habitat (APH) are CEA systems being used on the ISS. Veggie is a small plant growth chamber that can control temperature, lighting, and humidity. The system utilizes “plant pillows” which contain a ceramic arcillite substrate, a controlled release polymer fertilizer, seeds, and a wicking tube that provides water to foam in the pillow that passively works its way to the seeds. The APH is a more automated system that also includes cameras and sensors that adjust conditions without the need for crew input. Unlike the Veggie’s passive water system, the APH uses a root module watering system that utilizes laminar flow for water and nutrient delivery. To date, the Veggie system, the APH, and its predecessors have successfully grown a wide variety of plants including fruits, vegetables, oilseeds, and cereals have been successfully harvested onboard multiple spacecraft including during the space shuttle missions and currently on the ISS. In addition, the EDEN ISS project is an Earth-based CEA system, currently deployed in Antarctica, that is a fully self-sustained greenhouse system which has successfully produced 268 kg of edible biomass during its 286 day operation. The Veggie and APH utilize a substrate-based growth media for water and nutrient delivery, while the EDEN ISS uses an aeroponics system. In aeroponics systems, roots are exposed to air and sprayed intermittently with water and nutrients. This will be the preferred system for future space missions as it requires no soil (root substrates), is more efficient with nutrient delivery, and utilizes less water compared to substrate-based methods.

[0143] Despite the success of these plant production systems, they are still dependent on materials such as fertilizer and rooting substrates from outside of the CEA system. For spacecraft, this means a rocket launch to resupply these materials to maintain continuous production of crops is required. While the cost of launching a payload into LEO has decreased substantially thanks to advances in reusable rockets, the price can still range from $1,200 (SpaceX) to $10,000 (Rocket Lab) per pound. This cost is substantially larger for resupply missions beyond LEO. In the next few decades, the combination of NASA’s Artemis missions and the human-occupied commercial efforts in LEO will mean that more people than ever will be working in space at the same time. This creates the potential for long commercial queue times for payloads which could delay resupply missions of essential items, including food and plant growth materials. Hence, the need to make spacecraft more self-sustainable and less reliant on Earth-based materials is critical to ensure the success of both commercial and NASA operated spaceflight efforts.

[0144] This study, Dust Recycling for Environmental Agriculture Materials in Space (DREAMS), aims to further close the sustainability loop of CEA systems by using Dust from Astronauts and Space Habitats (DASH) as a root substrate and nutrient source for plant growth. DASH is generated onboard the ISS through the everyday activities of the crew consisting of biological and chemical elements shed from the occupants as well as built environment materials. It is shown herein that DASH contains a diverse fungal and bacterial community that can potentially be leveraged by plants in beneficial ways (FIG. 2A and FIG. 2B). For example, many fungi can support plant growth by biocontrol of pathogens with antibiotic production or competition as well as strengthen defense mechanisms of the plants themselves. Plant growth also requires nutrients such as potassium, calcium, magnesium, phosphorous, and nitrates to name a few. House dust samples from Earth-based homes have previously been shown to contain at least some of these nutrients (FIG. 3A). The current study on ISS dust also measured and found some nutrients (FIG. 3B).

[0145] DASH on the ISS is a waste product that is cleaned and collected weekly by the crew. It is vacuumed from the HEPA filter coverings that are part of the air ventilation system, which is where most of it is deposited. These vacuum bags are then stored onboard, which takes up valuable storage volume, until it can be burned up in the atmosphere. Finding a useful purpose for this otherwise waste product, such as for plants production, is advantageous.

[0146] The primary objective of DREAMS is to determine that DASH waste can be repurposed as a nutrient source and root substrate for plant production. Dust from the ISS HEPA filtration covers are obtained and used to grow red romaine lettuce (Lactuca .saliva), which is referred to as ‘Outredgeous’ for the remainder of the proposal. Outredgeous was chosen because of its reliable seed germination, rapid growth, naturally low microbial levels, nutritional content, and palatability. In addition, many studies have used Outredgeous, including on the ISS, which can aid in comparison of plants grown using DASH. A full microbial profile of DASH and plant samples are performed to understand the quantity, type, and functional ability of microorganisms present. Chemical analyses will be performed on all samples to look for nutrients relevant to plant growth in DASH and in edible portions of Outredgeous to determine nutritional content as well as presence of any hazardous elements. Outredgeous plant health is assessed using common physical characteristics such as leaf area, plant height, crop yield, and stomata density. Safety for human consumption are evaluated using established NASA guidelines. The results of this study can improve sustainability of food production on future spacecraft in and beyond LEO. In addition to the space-based applications, it can have direct applications on Earth by utilizing dust as an alternative nutrient source in areas of the world that have limited resources and high levels of food insecurity.

[0147] The overall goal of DREAMS is to determine if DASH can be repurposed as a nutrient source and / or root substrate for plant production in spacecraft. Outredgeous is grown in a plant pillow using DASH substrate and with a soluble nutrient solution extracted from DASH for use in an aeroponics system. Three key components of DASH and Outredgeous plants were evaluated: 1) microbiology, 2) chemical composition, and 3) plant health, nutrition, and safety.

[0148] Microbial analyses of DASH and Outredgeous.

[0149] A full microbial profile of dry DASH used as a root substrate and nutrient solution extracted from DASH is determined before, as well as after, use for plant growth. In addition, Outredgeous microbial components is measured in the phylloshphere (roots, stems, and leaves) and pseudo-rhizoshpere (usually soil, but in this case DASH). Analyses include quantification and identification of fungi as well as bacteria. A metatranscriptomic analysis of microbial RNA is performed to highlight metabolic pathways including those relevant to plant health, nutrient cycling, and human health.

[0150] Determine chemical composition of DASH and Outredgeous.

[0151] DASH is analyzed for chemical composition, specifically nutrients relevant for leafy green plant growth. This is done on the original dust samples from the ISS vacuum bag and in the nutrient solution extracted from DASH. Outredgeous leaves are evaluated for nutritional content as well as potentially hazardous compounds such as heavy metals.

[0152] Evaluate Outredgeous plant health, nutrition, and safety for human consumption.

[0153] The overall health of Outredgeous is evaluated using a variety of qualitative observations such as crop yield, plant height, leaf area, and stomata density. Nutritional content is assessed by measuring antioxidant activity and nutritional content in Outredgeous leaves. To determine if Outredgeous is safe for human consumption, the microbial content following established NASA requirements for ISS non-thermostabilized food requirements were evaluated.

[0154] Methods

[0155] Methodology and Approach-DASH collected from the ISS HEP A filter coverings which are part of the air ventilation system. DASH is provided by the NASA Johnson Space Center Toxicology and Environmental Chemistry group which is then sent to the Ohio State University to perform the experiments in this study. DASH is used to cultivate Outredgeous plants for two planting methods: 1) using DASH in a plant pillow as a root substrate / nutrient source and 2) extracting soluble nutrients from DASH for use in an aeroponics system. Microbial and chemical components of DASH as well as Outredgeous are characterized. Plant health, nutritional content, and safety for human consumption will also be determined.

[0156] Plant Growth Protocols-All planting and cultivation are performed at the Ohio Controlled Environment Agriculture Center at the Ohio State University. Two methods are tested that utilize DASH for growing red romaine lettuce (Lactuca saliva.jcv. ‘Outredgeous’). Planting method 1 uses DASH as both a nutrient source and a root substrate in the established plant pillow protocol. DASH is taken directly from the ISS vacuum bag for use as a replacement for the ceramic arcillite substrate and controlled release polymer fertilizer that is typically used in the plant pillow protocol. This method utilizes scenarios that include DASH only as well as a 1 : 1, 2: 1, 3 : 1, and 4:1 mixture of DASH / acrillite. no fertilizer is added to any of the scenarios using DASH and the specific quantity are determined based on the density of DASH received. A positive control is included that uses the standard Veggie hardware plant pillow mixture that has previously successfully grown Outredgeous. Planting method 2 extracts soluble nutrients from DASH for use in an aeroponics system. DASH is submerged in DI water and placed on a shaker table for ~12 hours at which point it is put through a 0.45pm filter. The collected solution is used as the water / fertilizer in the aeroponics system at a IX, 5X, 10X, 50X, and 100X dilution series. A positive control with the nutrient solution recipe from the EDEN ISS project is also used. The nutrient solution is sprayed onto the root zones for 30 seconds every 6 minutes and a positive control with a pre-made nutrient solution from the EDEN ISS project is used. A total of 72 plants are grown that includes 6 plants for each scenario from both planting methods. All plants are harvested and sampled after a total growth period of 30 to 40 days. An overview of planting methods, scenarios, and specific growth conditions can be found in FIG. 4.

[0157] Microbial analyses of DASH and Outredgeous-DNA is extracted from DASH and Outredgeous using a Maxwell® RSC PureFood GMO Authentication and Plant DNA kit respectively, with a modified extraction protocol. Dry DASH samples and nutrient solutions extracted from DASH are measured for all planting methods and scenarios before as well as after cultivation. After Outredgeous is harvested microbial analyses on leaf, root, and stem samples are performed. For all planting scenarios using DASH, a negative control is used which does not contain Outredgeous seeds. Quantification of total fungi bacteria is completed using qPCR on an Applied Biosystems Quantstudio 6 Flex and analyzed using Quantstudio Real-Time PCR Software vl.3. Total fungal concentration is measured using Aspergillus fumigatus as a standard and universal fungal primer set targeting the ITS region. Total bacterial concentrations are measured using a Bacillus atrophaeus standard and a primer / probe set targeting the 16S rRNA gene. Microbial composition are analyzed using DNA amplicon sequencing for ITS (fungal) and 16S (bacterial) regions as previously described. Processing sequence reads and assigning taxonomy utilize a well-established bioinformatics pipeline incorporating QIIME2, BLAST, and FHiTINGS. Microbial function is measured by performing a metatranscriptomic analysis on a subset of samples from DASH and Outredgeous. Specific samples are determined during study that reflect plants that grew well and those that did not. RNA is extracted using a Maxwell® RSC Plant RNA kit and sent to the Yale Center for Genomic Analysis for sequencing. An Illumina NovaSeq 2x100 lane with 5 million reads per sample is used for sequencing of bacterial, fungal, and plant RNA. Trinity pipeline with DESeq2 was used to create differentially expressed transcripts and clusters according to gene expression profiles. For these analyses, contigs from CD-HIT-EST is annotated with Trinotate, a suite designed for functional annotation of de novo assembled transcriptomes. Common protein domains are identified using BLAST+ with Swiss- Prot and PF AM databases. Using the SQLite database, a Trinotate annotation report is generated to produce Gene Ontology (GO) and KEGG Ontology (KO) terms. This data is mined for processes related to nutrient cycling, virulence factors, and plant metabolites which can help to understand what is happening during the cultivation of Outredgeous.

[0158] Elemental composition of DASH and Outredgreous-Chemical composition in DASH are evaluated in dry dust samples (Planting Method 1) and extracted nutrient solutions (Planting Method 2). These samples are measured for nutrients relevant for leafy crop growth (N, P, S, etc.) and potential growth inhibitors such as aluminum. Outredgeous stems, leaves, and roots are measured to determine how much nutrient uptake is occurring from each planting method and scenario. A modified EPA method 3050B is used for nutrient specific element extractions on dry DASH and lettuce leaves. In addition, the edible portion of Outredgeous leaves is measured for chemical hazards which may be harmful to human health such as heavy metals. Heavy metals in leaves are extracted using the U.S. Geological Survey digestion procedure. Specific heavy metal elements were chosen based on their risk to human health and because they have been previously found in ISS dust samples. Elemental analyses are performed using a Agilent Technologies 5100 Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). All measurements on ICP-OES will utilize a multi-element standard solution containing all elements of interest.

[0159] Plant Health, Nutrition, and Safety- The overall health of Outredgeous plants is evaluated by measuring plant height, leaf area, stomata density, and crop yield. Leaf area and plant height are measured using photographs and scanned images with Imaged software. Stomata density is measured from an epidermal impression sample of the Outredgeous leaves and counted using a 40X objective on a light microscope. Crop yield is determined by calculating the total fresh weight of Outredgeous harvested divided by the growing area for each plant. In addition, to the nutrient analyses on leaves described above, the antioxidant activity of Outredgeous leaves is measured. Antioxidant activity is a good indicator of nutritional value of produced lettuce for its natural anti-inflammatory benefits to human health. Antioxidant activity is determined by measuring total phenolic content and use of a Oxygen Radical Adsorption Capacity (ORAC) assay as previously described.

[0160] The NASA baseline microbial assessment is used to determine safety for human consumption. This includes measuring total viable microorganisms using an aerobic plate count, total yeast, and total mold assays. In addition, specific human pathogens are directly measured which will include Enter obacteriaceae, E. coli, Salmonella sp., and Aspergillus flavus. All protocols for microorganism enumerations and standards for safety are assessed using the Food and Drug Administration Bacteriological Analytical Manual. This microbial data combined with the heavy metal analyses described above can help determine the safety for human consumption of Outredgeous grown with DASH. Statistical Analyses-Chemical quantities, microbial counts, and plant health metrics are compared for each planting method and scenario within that method (e.g., different dilutions for nutrient solution and ratios for plant pillow) using a Kruskal-Wallis one-way ANOVA test. Microbial beta diversity differences are compared using the Bray-Curtis (for fungi) and UniFrac (for bacteria) distance matrices while alpha diversity are compared using richness as well as Shannon diversity metrics. To analyze statistical significance between microbial taxa for each planting method and scenario, a multiple comparisons test in Statistical Analysis System (SAS) MULTTEST with the false discovery (FDR) which has been described previously is used. For metatranscriptome analyses, differentially expressed transcripts based on the most significant FDR and fold-changes are clustered according to their patterns. Pairwise differential gene expression and co-expressed gene clusters relating to each planting condition and scenario are discovered using Trinity pipeline. Pairwise comparisons are performed for each planting method and scenario.

[0161] Results

[0162] DASH have sufficient nutrients to sustain and promote growth of Outredgeous plant in CEA growth chambers for both plant pillow and aeroponics planting methods. The goal was to accomplish use of DASH with as little alterations needed as possible. This is why the different dilutions for extracted nutrient solution and ratios of dust to acrillite are tested (FIG. 4). It has been previously reported that fungal plant pathogens such as Fusarium oxysporum are found onboard the ISS and have caused crop failures in orbit. DASH likely contains a vast amount of chemicals that are associated with the ISS built environment and human occupants that may inhibit or prevent crop growth. Some lettuce species are capable of phytoremediation of contaminants such as heavy metals in soils, but it is not known if Outredgeous is capable of phytoremediation from a DASH substrate if toxic metals are present. While pathogenic microorganisms have been found in DASH, it is expected that once water is introduced these will pose a minimal risk due to competition from non-pathogenic species and difficult mobility to edible portions of Outredgreous leaves.

[0163] Implications for Space- and Earth-Based Applications-The benefits of plant production on spacecraft go beyond food sources. For example, plant growth can also aid in converting carbon dioxide into breathable oxygen as well as improve the mental health of the crew through the process of farming and by introducing greenery in the spacecraft environment. Currently, plant growth on spacecraft is completely dependent on the resupply of important materials such as fertilizers and root substrates from Earth. The ability to utilize DASH as a nutrient source and / or root substrate for plant production onboard spacecraft is a novel approach to repurpose a waste product into a critical item required for mission success (FIG. 5). This promotes further self- sufficiency and sustainability of spacecraft which is essential for future missions that go beyond LEO. Reusing materials onboard spacecraft during long-duration missions can save time and money by reducing the amount of resupply launches needed from Earth. The results can also be incorporated into closed systems currently being developed to minimize supplies from outside sources such as Bioregenerative Life Support Systems. In addition to space-based applications, these methods can be used on Earth in areas of the world that have limited resources and experience food insecurity. Commercial buildings can also use already collected dust from vacuums to grow plants in their indoor and outdoor environments creating another metric for green building certifications. Understanding how waste products such as DASH can be recycled for useful applications promote sustainability goals in space and on Earth.

[0164] Other novel use cases for DASH in spacecraft include, but are not limited to, 3D printing materials, breaking down natural fibers (from clothing) or energy production (microbial fuel cells). Another use case for DASH is as a monitoring source to assess overall health of a spacecraft for potential hazards (chemical and microbial) that may be more comprehensive while requiring less time compared to surface swabs and air sampling.

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Claims

WHAT IS CLAIMED IS:

1. A method of cultivating plants using dust, comprising: a) collecting dust from closed-system environments; b) preparing a dust-based growth medium; and c) growing plants in the prepared dust-based medium.

2. The method of claim 1, wherein the dust-based growth medium comprises root substrate or nutrient solution.

3. The method of claim 1, wherein the closed-system environments comprise residential, commercial, or spacecraft.

4. The method of claim 1, wherein the dust comprises biological and chemical elements.

5. The method of claim 4, wherein the biological elements comprise beneficial fungi and bacteria for plant growth.

6. The method of claim 4, wherein the chemical elements comprise potassium, calcium, magnesium, phosphorous or nitrates.

7. The method of any of claims 1-6 , wherein the plants comprise radish, lettuce, collard greens, brussels sprouts, or cauliflower.

8. A method of measuring nutritional content of a plant using dust as a root substrate, comprising: a) collecting dust from closed-system environments; b) analyzing beneficial microorganisms in the dust; c) cultivating plants in the dust; and d) measuring nutritional content of the cultivated plants in the dust.

9. The method of claim 8, wherein the nutritional content comprises measuring antioxidant activity in the cultivated plants.

10. The method of claim 8, wherein the closed-system environments comprise residential, commercial, or spacecraft.

11. The method of claim 8, wherein the dust is further combined with inert substrates; wherein the inert substrates is acrillite.

12. A method of extracting nutrients from dust for plant growth, comprising: a) collecting dust from closed-system environments; b) suspending collected dust in water to release soluble nutrients; c) filtering the suspension of collected dust; d) obtaining filtered nutrient solution; and e) using the filtered nutrient solution in a hydroponic system.

13. The method of claim 12, wherein the closed-system environments comprise residential, commercial, or spacecraft.

14. A method for dual -use dust recycling in agriculture, comprising: a) collecting dust from closed-system environments, wherein the closed-system environments comprise residential, commercial, or spacecraft; b) utilizing collected dust as a physical substrate for plant roots; or extracting nutrients from the collected dust for use in hydroponic nutrient solutions.

15. The method of claim 14, wherein the nutrients are extracted from the collected dust by suspending collected dust in water and filtering the suspension.

16. The method of claim 14, wherein the physical substrate comprises collected dust combined with acrillite.

17. The method of claim 14, wherein the closed-system environments comprise residential, commercial, or spacecraft.

18. A controlled environment agriculture (CEA) system, comprising: a) a dust collection module for gathering Dust from Astronauts and Space Habitats (DASH); b) a dust processing unit to extract nutrients and remove hazardous elements from DASH; c) a plant growth module comprising a plant pillow system or an aeroponics system, wherein the plant pillow system serves as a root substrate; wherein the aeroponics system serves as a nutrient source; and d) a microbial analysis unit to monitor microbial communities within DASH and plant growth.

19. The CEA system of claim 18, further comprises an environmental condition module, wherein the environmental condition module comprises ideal temperature, relative humidity, carbon dioxide and lighting for plant growth.

20. The CEA system of claim 19, wherein the temperature is 24°C-28°C.

21. The CEA system of claim 19, wherein the relative humidity is 70%-75%.

22. The CEA system of claim 19, wherein the carbon dioxide is 1200ppm.

23. The CEA system of claim 19, wherein the lighting is 16-hour photoperiods at 150 mol / m3-200 mol / m3.

24. The CEA system of claim 18, wherein the plant pillow system consists of collected dust mixed with a ceramic arcillite substrate, a polymer fertilizer, seeds, and a wicking tube.

25. The CEA system of claim 18, wherein the aeroponics system comprises soluble nutrient solution derived from the DASH.

26. The CEA system of claim 18, wherein the dust processing unit comprises: a) a nutrient extraction subsystem to solubilize nutrients, wherein the nutrients comprise potassium, calcium, magnesium, or phosphorus from the DASH; and b) a heavy metal removal subsystem, wherein the heavy metals comprise lead, mercury, cadmium or chromium.

27. The CEA system of claim 18, wherein the microbial analysis unit comprises: a) identify and quantify beneficial and harmful microorganisms in DASH; and b) monitor changes in plant-associated microbiomes during plant growth.

28. A method of utilizing DASH in spacecraft agriculture, comprising: a) collecting DASH from onboard filtration system; b) processing DASH to extract nutrients; c) utilizing the processed DASH as a root substrate in a plant pillow system or as a nutrient source in an aeroponics system; d) cultivating plant in the plant pillow or the aeroponics system; and e) monitoring plant growth, health, and safety for human consumption.

29. The method of claim 28, wherein the cultivated plant is Lactuca sativa (red romaine lettuce).

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