# US 11,306,012 B2 · Soil-based flow-through rhizosphere system for treatment of contaminated water and soil

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- Number: US 11,306,012 B2
- Document ID: US11306012B2
- Status: Granted
- Kind: B2
- Application number: 16/490,735
- Filed: 2018-12-06
- Published: 2020-01-02
- Granted: 2022-04-19
- Earliest claimed: 2018-01-02
- Assignees: Reed Scientific Services Ltd.
- Applicants: Reed Scientific Services Ltd.
- Inventors: Lucian Stephen Gill
- Web page: https://patentgrep.com/patent/US11306012B2

## CPC

- C02F 3/34: characterised by the microorganisms used
- B09C 1/08: chemically
- B09C 1/10: microbiologically, {biologically} or by using enzymes
- B09C 1/105: {using fungi or plants}
- C02F 3/006: {Regulation methods for biological treatment}
- C02F 3/046: {Soil filtration}
- C02F 3/327: {characterised by animals and plants}
- C02F 3/348: {characterised by the way or the form in which the microorganisms are added or dosed}
- C09K 17/00: Soil-conditioning materials or soil-stabilising materials (specially adapted for boreholes or wells C09K8/00; fertilisers C05; consolidating by placing solidifying or pore-filling substances in the soil E02D3/12)
- C12N 1/20: Bacteria; Culture media therefor
- B09C 2101/00: In situ
- C02F 2101/32: Hydrocarbons, e.g. oil
- C02F 2101/36: containing halogen
- C02F 2103/06: Contaminated groundwater or leachate
- C02F 2203/004: comprising a selector reactor for promoting floc-forming or other bacteria
- C02F 2301/08: Multistage treatments, e.g. repetition of the same process step under different conditions
- Y02W 10/10: Biological treatment of water, waste water, or sewage
- Y02W 10/37: using solar energy
- C02F: TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE (separation in general B01D; special arrangements on waterborne vessels of installations for treating water, waste water or sewage, e.g. for producing fresh water, B63J; adding materials to water to prevent corrosion C23F; treating radioactively-contaminated liquids G21F9/04; regeneration of reactants for recirculation into processes, see the relevant places for the processes)
- B09C: RECLAMATION OF CONTAMINATED SOIL (gatherers for removing stone or the like from the soil A01B43/00; sterilising soil by steam A01G11/00; removing undesirable matter, e.g. rubbish, from the land E01H15/00)
- C09K: MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02W: CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT

## Abstract

A process for constructing a soil-based rhizosphere flow-through system to break down contaminants in contaminated water. The process includes the steps of: providing plants planted in soil in a test bioreactor, the plants providing a rhizosphere; exposing the rhizosphere to the contaminated water; extracting microorganisms from the rhizosphere following their exposure to the contaminated water; preparing a microbial suspension from the extract; subjecting the microbial suspension to growth conditions to increase the concentration of the microorganisms, thereby preparing a soil conditioner; adding the soil conditioner to soil in a contained area having a water flow inlet and outlet; and planting a plurality of plants in the soil, the plants being of the same species as the plants of the test bioreactor.

## Claims

### Claim 1

A process for preparing a soil conditioner to increase the rate of decontamination of contaminated water or contaminated soil, the process comprising:
a) providing a rhizosphere;
b) exposing the rhizosphere to the contaminated water;
c) obtaining a sample of the rhizosphere, the sample comprising plant roots and soil surrounding the rhizome;
d) extracting microorganisms from the sample after step c) to prepare a microbial extract;
e) preparing a microbial suspension from the microbial extract; and
f) subjecting the microbial suspension to growth conditions to increase the concentration of the microorganisms, thereby preparing the soil conditioner, wherein the soil conditioner includes microorganisms originally present in the contaminated water and microorganisms originally present in the rhizosphere.

### Claim 2 (depends on 1)

The process of claim 1, wherein step b) comprises generating a flow of the contaminated water across the rhizosphere, the flow entering at an inlet and exiting at an outlet.

### Claim 3 (depends on 2)

The process of claim 2, wherein the flow of the contaminated water is conducted for a period of about 4 to about 8 weeks.

### Claim 4 (depends on 2)

The process of claim 2, further comprising testing of outflow from the outlet to identify one or more metabolic products of one or more contaminants present in the contaminated water.

### Claim 5 (depends on 1)

The process of claim 1, further comprising measuring the amount of microorganisms present in the extract in the microbial suspension.

### Claim 6 (depends on 1)

The process of claim 1, further comprising measuring the amount of microorganisms present in the soil conditioner, wherein a plate count of at least about 106 counts/mL indicates that the soil conditioner is suitable for use in treatment of a soil bed in construction of the soil-based flow-through rhizosphere system.

### Claim 7 (depends on 1)

The process of claim 1, wherein the rhizosphere is of a wetland plant.

### Claim 8 (depends on 7)

The process of claim 7, wherein the wetland plant is selected from the group consisting of Phragmites australis, Arundo donax L., Neyraudia reynaudiana, Phalaris arundinacea, Glyceria maxima, Elegia tectorum, Thamnochortus insignis, a species of the genus Calamagrostis, a species of the genus Sparganium, and a species of the genus Typha.

### Claim 9 (depends on 1)

The process of claim 1, wherein steps a) and b) are conducted in a test bioreactor having a volume of about 10 L to about 50 L.

### Claim 10 (depends on 1)

The process of claim 1, wherein the contaminated water comprises organic contaminants.

### Claim 11

A process for constructing a soil-based rhizosphere flow-through system to break down contaminants in contaminated water, the process comprising:
a) providing plants planted in a test bioreactor, the plants providing a rhizosphere;
b) exposing the rhizosphere to the contaminated water;
c) obtaining a sample of the rhizosphere, the sample comprising plant roots and soil surrounding the rhizome;
d) extracting microorganisms from the sample after step c) to prepare a microbial extract;
e) preparing a microbial suspension from the microbial extract;
f) subjecting the microbial suspension to growth conditions to increase the concentration of the microorganisms, thereby preparing a soil conditioner, wherein the soil conditioner includes microorganisms originally present in the contaminated water and microorganisms originally present in the rhizosphere;
g) adding the soil conditioner to different soil in a contained area having a water flow inlet and outlet; and
h) planting a plurality of plants in the different soil, the plants being of the same species as the plants of step a).

### Claim 12 (depends on 11)

The process of claim 11, wherein step b) comprises generating a flow of the contaminated water across the rhizosphere, the flow entering at an inlet and exiting at an outlet.

### Claim 13 (depends on 12)

The process of claim 12, wherein the flow of the contaminated water is conducted for a period of about 4 to about 8 weeks.

### Claim 14 (depends on 12)

The process of claim 12, further comprising testing of outflow from the outlet to identify one or more metabolic products of one or more contaminants present in the contaminated water.

### Claim 15 (depends on 11)

The process of claim 11, further comprising measuring the amount of microorganisms present in the extract in the microbial suspension.

### Claim 16 (depends on 11)

The process of claim 11, further comprising measuring the amount of microorganisms present in the soil conditioner, wherein a plate count of at least about 106 counts/mL indicates that the soil conditioner is suitable for use in treatment of a soil bed in construction of the soil-based flow-through rhizosphere system.

### Claim 17 (depends on 11)

The process of claim 11, wherein the rhizosphere is of a wetland plant.

### Claim 18 (depends on 17)

The process of claim 17, wherein the wetland plant is selected from the group consisting of Phragmites australis, Arundo donax L., Neyraudia reynaudiana, Phalaris arundinacea, Glyceria maxima, Elegia tectorum, Thamnochortus insignis, a species of the genus Calamagrostis, a species of the genus Sparganium, and a species of the genus Typha.

### Claim 19 (depends on 11)

The process of claim 11, wherein steps a) and b) are conducted in a test bioreactor having a volume of about 10 L to about 50 L.

### Claim 20 (depends on 11)

The process of claim 11, wherein the contaminated water comprises organic contaminants.

### Claim 21 (depends on 1)

A soil-based flow-through rhizosphere system for breaking down contaminant organic compounds in contaminated water, the system comprising:
a) a contained area comprising soil treated with a soil conditioner prepared according to the process of claim 1 and planted with plants of the same species as the rhizosphere, the contained area having a water flow inlet and a flow outlet;
b) a tank for holding the contaminated water before entry of the contaminated water into the contained area; the tank in flow communication with the flow inlet; and
c) a retention pond in communication with the flow outlet.

### Claim 22 (depends on 21)

The system of claim 21, further comprising a soak-away discharge pond in flow communication with the retention pond to allow discharge of treated water into the environment.

### Claim 23 (depends on 21)

The system of claim 21, further comprising a recirculation line in flow communication between the retention pond and the water flow inlet.

### Claim 24 (depends on 21)

The system of claim 21 wherein the contained area is a container configured to provide predominately horizontal flow of the contaminated water from the flow inlet to the flow outlet.

### Claim 25 (depends on 24)

The system of claim 24, wherein the container comprises a soil depth of between about 0.4 m to about 0.5 m to provide a predominately aerobic soil environment.

### Claim 26 (depends on 21)

The system of claim 21 wherein the contained area is a container configured to provide predominately vertical flow of the contaminated water from the flow inlet to the flow outlet.

### Claim 27 (depends on 26)

The system of claim 26, wherein the container comprises a soil depth of between about 0.8 m to about 1.0 m to provide soil having both a predominately aerobic soil environment closer to the soil surface and a predominately anaerobic soil environment closer to the bottom of the container.