Biomolecular zonal compositions and methods

Abstract

A composition is provided including biospheres and bacteria, wherein the biospheres are cellulosic biopolymers and sized in the range from 500 nanometers to 80 microns and have a minimum free swell absorption capacity of 400 times by weight and a maximum free swell absorption capacity of 1200 times by weight, wherein the composition is formable as a gelatinous matrix.

Claims

1. A composition comprising a blend of rechargeable biospheres and water, and optionally bacteria, biological and/or chemical reagents, wherein the biospheres are cellulosic biopolymers and have a free swell absorption capacity, wherein the composition is formable into an environmentally responsive gelatinous matrix which delivers and sustains bacteria, biological and/or chemical reagents in situ.

2. The composition of claim 1, wherein the biospheres gradually release moisture.

3. The composition of claim 1, wherein the viscosity of the composition is from 6 centipoise (cps) to 217 centipoise (cps).

4. The composition of claim 1, wherein the viscosity of the composition is from 217 centipoise (cps) to 1,159 centipoise (cps).

5. The composition of claim 1, wherein the viscosity of the composition is from 1,236 centipoise (cps) to 5,021 centipoise (cps).

6. The composition of claim 1, wherein the viscosity of the composition is from 5,021 centipoise (cps) to 47,311 centipoise (cps).

7. The composition of claim 1, wherein the biospheres are sized in the range from 500 nanometers to 80 microns.

8. The composition of claim 1, wherein the biospheres have a maximum free swell absorption capacity of 1200 times by weight.

9. The composition of claim 1, wherein the biospheres have a minimum free swell absorption capacity of 400 times by weight.

10. A method for bioremediation in situ, the method comprising: preparing a blend of liquid bacterial culture with biospheres, wherein the biospheres are cellulosic biopolymers and sized in the range from 500 nanometers to 80 microns and have a minimum free swell absorption capacity of 400 times by weight and a maximum free swell absorption capacity of 1200 times by weight; applying the blend at a site in need of bioremediation; and forming a gelatinous matrix with the blend.

11. A method for delivering and/or hosting biological and/or chemical reagents in a gelatinous matrix, the method comprising: obtaining biospheres which are cellulosic biopolymers and sized in the range from 500 nanometers to 80 microns and have a minimum free swell absorption capacity of 400 times by weight and a maximum free swell absorption capacity of 1200 times by weight; mixing the biospheres with a bacterial and/or chemical reagent to form a mixture; and forming a gelatinous matrix with the mixture.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 reports results from treatment with average hot spot concentrations (>5,000 mg/kg);

(2) FIG. 2 reports volumetric projections; and

(3) FIG. 3 reports results of a CUP analysis after 14 and 28 weeks.

DETAILED DESCRIPTION

(4) In all these novel applications and treatment applications, both field and bench-scale studies, have demonstrated the ability of the blends with biospheres to enhance the process of biodegradation. As a result, many of the active properties that combine to produce these highly productive outcomes also distinguish the in-situ technology with biospheres from conventional bioremediation.

(5) One particularly important distinction is the capacity to establish and sustain microbial activity at levels where potent micellar bio-surfactant solutions naturally occur and assist organic molecules to dissolve, mix and interact with the selected bacteria to simplify application and optimize the process of biodegradation.

(6) These optimized, naturally occurring, environmentally safe biological catalysts disrupt the complex molecular chains of hydrocarbon based contaminants and, this process within the gelatinous matrix created by the blend with biospheres, produces more easily digestible molecules that become encapsulated, together with the bacteria, in cores of nano-sized micelles that sustain favorable contact with the water that surrounds them and, thus, provide an ideal microenvironment for optimizing the reaction kinetics associated with successful in-situ biodegradation.

(7) Therefore, the blend with biospheres provides an optimized bioremediation process in which nano-sized micelles are created, while typically no micelles are usually created in a typical bioremediation process with liquid bacterial culture. This distinction is important because of the following significant advantages, which become apparent, when comparing the key characteristics of using the blend with biospheres rather than a conventional liquid culture to perform in-situ biological treatments.

(8) Water's natural ability to freely migrate through the soil and flow across its surface, is as essential for life as it is wasteful and impractical when being used as the primary vehicle for distribution of specific treatments to enrich or remediate the soil. Therefore, with no alternative to water, other than the way it comes out of a tap, for the distribution of specialist biodegrading bacteria, in-situ bioremediation remains a highly unpredictable and intensive procedure. In complete contrast, the blend with biospheres augments water, and while it remains fluid and continuous, it also becomes a controllable life optimizing gelatinous host, which is organic with a cellular matrix to optimize the potential for biological life cycles to thrive.

(9) Despite being a much less intensive process, the gelatinous matrix, which is rechargeable in-situ, is also a far more predictable inoculant than conventional liquid cultures. The blend with biospheres combines absorption, rapid capture and controlled release, within a microbial inoculant, transforms the water element into an optimizing super carrier that can act as a biodegradable subterranean sink, to sustain moisture levels. The gelatinous matrix makes maintaining sufficient moisture and, consequently, in-situ remediation, much more efficient.

(10) Further and also in complete contrast to conventional liquid cultures, the blend with biospheres establishes a natural nutritious reservoir in the ground or across any surface when it is applied. This presents a major advance as the novel properties, within this gelatinous reservoir, of rechargeable super absorption, rapid capture and slow release combine to help even out the unpredictability associated with microbial survival. The most critical stages being at the start of a project, due to contamination causing high levels of toxicity, and towards the end of the process, when a lack of food occurs as a result of the land becoming clean again.

(11) Another significant advance, is controlled migration through dynamic viscosity management so a gelatinous matrix with biospheres and bacteria can radiate through the soil more slowly. This is important as additional control maximizes the opportunity for bacteria to attach themselves to the target food source, which is the contaminant to be removed from the soil or water source. This major difference creates the possibility for establishing billions more Colony Forming Units (CFUs) far more quickly, thus, making the whole procedure faster and much more predictable.

(12) Another advantage, from what occurs within the gelatinous matrix with biospheres, is helping air to flow through the soil by creating micro-pressures as it expands and contracts in the ground. This positive influence over the process is a direct consequence of the cyclical process of capturing and releasing water and nutrients that sustain high levels of microbial activity to, ultimately, optimize the process of in-situ bioremediation.

(13) In at least some embodiments, blends with biospheres are formulated to act as a self-buffering system to independently maintain the correct pH value in the soil throughout the entire treatment process. Moreover, the exceptionally high retention characteristic within blends with biospheres results in a far greater proportion of its pH buffering components remaining in position for much longer than would be possible when using a conventional liquid inoculant and, thus, the blend with biospheres optimizes the potential for these components to act as a continuous bacteria specific pH buffering stimulant.

(14) To prevail over the many limitations facing conventional in-situ bioremediation, the blend with biospheres transforms water to intensify targeting and interaction with contaminants, while still sustaining a healthy microenvironment that optimizes the potential for biological life cycles to flourish. This transformation massively increases the surface area that is made available for the bacteria to grow up on and, thus, the biomass that results is also increased exponentially.

(15) These advances are realized as organic micro-particles (biospheres) are meticulously blended with a water based liquid culture and optionally with other natural synergistic ingredients to develop both site and application specific embodiments.

(16) In some embodiments, the blend with biospheres can be further formulated with at least one component selected from Table 1.

(17) TABLE-US-00001 TABLE 1 Components for Heterotrophic Bacterium Growth. Minimum Amount Component (per liter) Function of Component Sodium Citrate (Na.sub.3C.sub.6H.sub.5O.sub.7) 10 g/1.0% C & Energy Source Ammonium Sulfate 1 g/0.1% pH buffer; N & P Source (NH.sub.4).sub.2SO.sub.4 Monosodium phosphate 2.5 g/0.25% pH buffer; P & K Source (NaH.sub.2PO.sub.4) Dipotassium Phosphate 2.5 g/0.25% pH buffer; P & K Source (K.sub.2HPO.sub.4) Magnesium Sulfate 0.207 g/ S & Mg.sup.++ Source (MgSO.sub.4); or Eprom Salt 0.0207% (MgSO.sub.4 7H.sub.2O) Ferrous Sulfate (FeSO.sub.4) 0.01 g/0.001% Fe.sup.++ Source

(18) In some embodiments, the blend with biospheres and other optional components discussed above, is obtained by using a vacuum induction system so that the biospheres are mixed with the water under intense sheer energy. This is essential as it increases the specific surface of the available liquid by several hundred thousand times and, thus, as the biospheres are separated momentarily, they become wetted and dispersed completely without forming any lumps through agglomeration.

(19) Finally, the blend can be further refined by low to medium rotation before being left to rest and bottling.

(20) Rechargeable in-situ, the resulting cellular microenvironment provides a surface area that has the capacity to establish and sustain microbial activity at levels where potent micellar surfactant solutions naturally occur and assist organic molecules to dissolve, mix and interact with the selected bacteria to simplify application and optimize the process of biodegradation.

(21) The blend with biospheres can be used with any microorganisms. At least in some embodiments, the microorganisms utilized are indigenous to the soil and the ocean, they are not genetically altered and fall within nonpathogenic homology groups. Such microorganisms may include any of the following: a)Pseudomonas putidaA gram negative rod that was isolated from fuel oil contaminated soil. This aerobic Pseudomonas falls within the non-pathogenic P. flourescens homology group; b)Acinetobacter johnsonii/genospecies 7A non-spore forming gram negative rod that was isolated from an Atlantic Ocean estuary off the coast of Hampton, N.H. These bacteria were selected for their ability to degrade crude oil and other petroleum hydrocarbons in marine environments; c)Alcaligenes faecallis Type IIA gram negative rod that was isolated from fuel oil contaminated soil. Alcaligenes faecallis Type II. These bacteria are not gram positive and, thus, they are not Staphylococci sp., Bacillus sp., or Streptococci sp. Biolog analyses also excluded Salmonella, fecal coliform and Shigella; d) Pseudomonas-unidentified fluorescentA gram negative rod that was isolated from fuel oil contaminated soil. This aerobic Pseudomonas falls within the non-pathogenic P. flourescens homology group.

(22) A person of skill would further appreciate that the blends with biospheres can be used in in-situ methods where precision delivery is needed.

(23) Further advantages of the blends with biospheres include the unique ability to engineer the dynamic viscosities of its gelatinous matrix, to slow down migration through the soil and stabilize the coverage of the matrix for prolonged periods across treated surfaces. This provides significant additional control that maximizes the opportunity for the bacteria to attach themselves to the target organic waste to be degraded or absorbed. Therefore, in complete contrast with conventional treatments, the present method minimizes wasting inoculant, while also helping to establish billions more bacteria far more quickly to make the process faster and much more predictable.

(24) In further embodiments, fluorescence can be added to a biosphere so that the migratory patterns and stability of a gelatinous matrix can be tracked in the field and observed. In these embodiments, samples can be analyzed under UV light and/or by UV microscopy.

(25) Further embodiments include kits which comprise a blend with biospheres. Such blends can be stored as a dry powder and mixed with water and a bacterial culture of choice prior to be used in the field.

(26) The blend with biospheres and gelatinous matrix it creates has the potential to improve many commercial practices in the areas of water conservation, diffuse pollution management, land remediation, restoration of soils and the maintenance remediation of construction materials.

(27) This invention will be further described by the way of the following non-limiting examples.

Example 1

(28) A large field trial was conducted utilizing a conventional liquid bacterial culture in Phase 1 as shown in FIG. 1. See cells 1 and 4 before phase 2 treatment.

(29) In Phase 2, both cells were treated with a blend comprising biospheres and a positive outcome was achieved. See FIG. 1, (Cells 2 & 3 not treated during Phase 1).

(30) Moreover, the graph in FIG. 1 also delineates a second set of identical patterns of TPH biodegradation. These results are also significant, as they occurred in the treatment areas designated as the control locations during the study and, therefore, neither area had received any treatment whatsoever before Phase 2.

(31) Ultimately, in these four heterogeneous cases, analysis had demonstrated identical patterns of biodegradation. Thus, the indicated change in TPH concentrations was due to biodegradation that resulted from the blend with biospheres in-situ remediation procedure carried out in the second phase of this environmental study. See FIG. 1. (Cells 2 & 3 not treated during Phase 1).

(32) Additional studies were conducted and demonstrate the overall reduction in contaminant mass that was achieved after the in-situ remediation procedure with a blend comprising biospheres that was completed in the second phase of the same environmental study. These results are reported in FIG. 2 alongside instructive comparative data taken from the conventional in-situ bioremediation treatments that were performed in Phase 1 of the study. These results also verify the only significant reduction in pollution that had occurred on the site, over a period of eight years of scientific monitoring, was due to the biodegradation that resulted from utilizing the blend with biospheres.

(33) Another distinction between conventional liquid culture and a blend with biospheres can be observed in shelf-life studies where random examples are taken at a specific point of final production and stored in separate twenty liter containers so that periodic samples can be taken for analysis to assess microbial viability over various periods of time. As shown in FIG. 3, various microbial cultures remained viable after 28 weeks in storage.

(34) Overall, after fourteen weeks, these laboratory results demonstrated three blends with biospheres had sustained strong viability, one blend had sustained moderate viability and the liquid culture, used as the control, had sustained only a low level of viability. See FIG. 3.

(35) After 28 weeks, the results demonstrated all four blends had sustained strong viability juxtaposed with the liquid culture that demonstrated no activity. See FIG. 3.

(36) The results from this study are particularly instructive because various blends with biospheres tested and the control were all produced from the same batch of liquid culture.

(37) While particular embodiments of the present biomolecular zonal compositions and methods have been described herein, it will be appreciated by those skilled in the art that changes and modifications may be made thereto without departing from the invention in its broader aspects and as set forth in the following claims.