CN110872566A

CN110872566A - Method and product for biodegrading waste

Info

Publication number: CN110872566A
Application number: CN201811203649.8A
Authority: CN
Inventors: 李清心; 吴金川; 大卫·陈; 方业如
Original assignee: Weisong Environmental Technology Shanghai Co ltd
Current assignee: Weibo Ecological Technology Shanghai Co ltd
Priority date: 2018-09-04
Filing date: 2018-10-16
Publication date: 2020-03-10
Also published as: TW202010836A; SG10201807584UA

Abstract

The invention provides a method for preparing a product for degrading waste, a microbial flora or mixed microbial composition, a product for degrading waste and a waste degradation method. The preparation method of the product for degrading the waste comprises the following steps: providing at least one isolated strain of a microorganism from excreta, providing at least one strain of a microorganism that produces lactic acid and/or acetic acid, providing at least one strain of yeast, providing at least one strain of Bacillus (Bacillus sp.), and combining the strains of microorganisms to produce a microbial flora for degrading waste.

Description

Method and product for biodegrading waste

Technical Field

The present invention relates generally to biodegradation of waste, and more particularly, to a method of preparing a product for degrading waste, a microbial flora or a mixed microbial composition for degrading waste, a product comprising the microbial flora or the mixed microbial composition, and a method of degrading waste.

Background

The amount of waste produced by humans continues to increase at a surprising rate. Almost any human activity produces waste. This waste may be in the form of solids, liquids or gases. The waste is produced by any organism.

Human waste includes, for example, municipal waste, sewage waste, and industrial waste. Appropriate waste management is increasingly important and there are many factors, aspects and goals to consider; such as the reduction/removal of toxic and harmful substances. Biodegradation of organic waste is an important part of the waste management community.

In areas where it may not be possible to use a toilet facility with a sewage treatment system and waste treatment, there is a need to develop efficient composting methods and products for the discharge of waste.

Disclosure of Invention

According to a first aspect, the present invention provides a method for preparing a product for degrading waste, comprising:

(i) providing at least one isolated microbial strain from an excreta;

(ii) providing at least one lactic acid and/or acetic acid producing microbial strain;

(iii) providing at least one yeast strain;

(iv) providing at least one strain of Bacillus, and

(v) (iii) combining the microbial strains from (i) to (iv) to produce a microbial population for degrading waste.

In a second aspect, the present invention provides a microbial flora or mixed microbial composition comprising:

(i) at least one isolated microbial strain from excrement;

(ii) at least one lactic acid and/or acetic acid producing microbial strain;

(iii) at least one yeast strain; and

(iv) at least one strain of Bacillus (Bacillus sp.).

In a third aspect, the present invention provides a product for degrading waste comprising a microbial flora or a mixed microbial composition according to the second aspect.

In a fourth aspect, the present invention provides a method of degrading waste. The method comprises the following steps: providing one of a microbial flora or a mixed microbial composition according to the second aspect and a product for degrading waste according to the third aspect; mixing one of a microbial flora or a mixed microbial composition and a product for degrading waste with the waste; biodegrading the waste with one of a microbial flora or a mixed microbial composition and a product for degrading the waste.

Other aspects and advantages of the present invention will become apparent from the following detailed description, taken in conjunction with the accompanying drawings, illustrating by way of example the principles of the invention.

Drawings

FIG. 1 is a photograph of a fixed microbial flora that may be used;

FIG. 2 is a photograph when human waste is added to a fixed microbial flora in a bio-toilet room for 0 hour (h); and

FIG. 3 is a photograph of the immobilized microbial flora completely degrading human waste after 24 hours.

Definition of

As used herein, "biodegradation" refers to decomposition (break down) or decomposition (decompose) by a biological process. Thus, the process of decomposing organic materials by contacting the materials with bacteria is one example of biodegradation.

As used herein, the term "composting" or "compostable" refers to the biodegradation or decomposition of organic matter and is carried out by various microorganisms, including bacteria, fungi, and the like. The compost product is compost, which contains a mixture of rotting organic matter. The compost can be used as a fertilizer.

As used herein, the terms "comprises" and "comprising" should be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components, or groups thereof. However, in the context of the present disclosure, the term "comprising" or "including" also includes "consisting of. Variations of the word "comprising", such as "comprises" and "comprising", and "including", such as "comprises" and "includes", have correspondingly varied meanings.

As used herein, "microorganism" is understood to refer to any microscopic organism, including eukaryotes, prokaryotes, or viruses; further included are, but are not limited to, bacteria (gram positive, gram negative or gram variable), fungi, viruses, protists, algae and reproductive forms thereof, including cysts and spores. With respect to bacteria, for example, there are included mycoplasma, rickettsia and chlamydia that replicate in eukaryotic cells, and those that do not replicate in eukaryotic cells. The term "microorganism" is used interchangeably with "microorganism" and "microorganism".

As used herein, the term "isolated" as applied to a microorganism refers to a microorganism that has been removed and/or purified from its naturally occurring environment. Thus, an "isolated strain" of a microorganism, as used herein, is a strain that has been removed and/or purified from its natural environment. Thus, an "isolated microorganism" does not include microorganisms that are present in the environment in which it naturally occurs. Furthermore, the term "isolated" does not necessarily reflect the degree of purification of the microorganism. A "substantially pure culture" of a microbial strain refers to a culture that is substantially free of other microorganisms than the desired microbial strain. In other words, a culture of a substantially pure microbial strain is substantially free of other contaminants (including microbial contaminants as well as undesirable chemical contaminants). Furthermore, as used herein, a "biologically pure" strain is intended to mean a strain that is separate from the material with which it is normally naturally associated. Note that strains associated with other strains, or strains associated with compounds or materials not normally found therein, are still defined as "biologically pure". Of course, a single culture of a particular strain is "biologically pure". As used herein, the term "enriched culture" of an isolated microbial strain refers to a microbial culture containing more than 50%, 60%, 70%, 80%, 90%, or 95% of the isolated strain.

As used herein, a microbial flora refers to a mixed population of two or more strains of microorganisms. Generally, microbial strains can be naturally occurring or combined together and achieve a particular purpose. For example, the combination of microbial populations of the invention is for biodegradation.

As used herein, the term "organic matter" (used interchangeably with organic material) includes any material comprising carbon, including fossil and non-fossil materials. Non-limiting examples of organic matter include biomass, lignocellulosic matter, and hydrocarbonaceous materials (e.g., lignite, oil shale, and peat).

As used herein, "waste comprising organic matter" includes biological waste, manure, green waste, municipal waste, sewage, food waste, and agricultural waste, as well as industrial organic waste. Manure may include manure produced by humans and various animals, including farm animals, such as cattle, sheep, horses, pigs, goats, rabbits, and poultry, such as chickens, turkeys, and ducks. Green waste may include various substrates from a variety of sources, such as yard waste, including grass clippings, tree, shrub, and fence trimmings, as well as foliage, as well as household and commercial food waste. Municipal waste may include residential and commercial waste, such as paper, wood, food, and yard waste. Biodegradable or compostable waste can be separated from mixed non-biodegradable matter. Sewage sludge can be used as a source of organic waste.

Detailed Description

(i) providing at least one isolated microbial strain from an excreta;

(iii) providing at least one yeast strain;

(iv) providing at least one strain of Bacillus, and

(i) at least one isolated microbial strain from excrement;

(ii) at least one lactic acid and/or acetic acid producing microbial strain;

(iii) at least one yeast strain; and

(iv) at least one strain of Bacillus (Bacillus sp.).

In a third aspect, the present invention provides an isolated microbial strain from excrement.

In a fourth aspect, the present invention provides a substantially pure culture of an isolated microbial strain from excrement.

Any isolated microbial strain derived from an excreta may be used to prepare the microbial flora, the microbial flora derived from an excreta or the isolated microbial strain used in the present invention. For example, an isolated microbial strain from excreta may be selected from the group consisting of Paenibacillus (Paenibacillus sp.), Bacillus (Bacillus sp.), and Pediococcus (Pediococcus sp.).

The Paenibacillus may comprise Paenibacillus DSMZ 32878. The bacillus may comprise bacillus DSMZ 32879. The genus Pediococcus may comprise Pediococcus acidilactici (Pediococcus acililicici). In particular, Pediococcus may comprise Pediococcus acidilactici DSMZ 32880. The DSMZ number throughout the description is the number of the DSMZ-German Collection of microorganisms (International preservation organization of the Budapest treaty) of the Rebnitz institute.

It is to be understood that the isolated microbial strain from excreta of any aspect of the present invention may consist of paenibacillus DSMZ32878, bacillus DSMZ 32879 and/or pediococcus acidilactici DSMZ 32880.

Accordingly, the present invention provides an isolated strain of bacillus sp. The invention also provides an isolated strain of bacillus DSMZ 32879. The invention also provides an isolated strain of pediococcus acidilactici DSMZ 32880.

Any lactic acid and/or acetic acid producing microbial strain may be used for preparing the microbial flora or for the microbial flora of the present invention. The lactic acid and/or acetic acid producing microbial strain may be an isolated lactic acid and/or acetic acid producing bacterium. For example, the lactic acid and/or acetic acid producing microbial strain may be selected from the group consisting of Acetobacter sp, Lactobacillus sp, and Lactococcus sp.

Acetobacter may include Acetobacter pasteurianus (Acetobacter pasteurianus). The genus Lactococcus may include Lactococcus lactis (Lactococcus lactis). It is to be understood that any commercially available Acetobacter, Acetobacter pasteurianus, Lactobacillus, lactococcus and lactococcus lactis may be used.

It is to be understood that in one embodiment, the lactic acid and/or acetic acid producing microbial strain may consist of acetobacter pasteurianus, lactobacillus and/or lactococcus lactis.

Advantageously, the at least one lactic acid and/or acetic acid producing microbial strain added creates an acidic environment to inhibit harmful microbes and odors, as well as to promote biodegradation.

Any yeast strain can be used to prepare the microbial flora or microbial flora for use in the present invention. The yeast strain may be an isolated yeast strain. For example, the yeast strain may be Saccharomyces (Saccharomyces sp.). The Saccharomyces genus may include Saccharomyces cerevisiae (Saccharomyces cerevisiae). It will be appreciated that any Saccharomyces cerevisiae may be used, for example, from the commercially available Baker's yeast.

Advantageously, the addition of at least one yeast strain reduces unpleasant odors.

Any strain of Bacillus may be used to prepare the microbial flora or microbial flora for use in the present invention. The bacillus strain may be isolated. The at least one strain of Bacillus used in (iv) of the first and second aspects of the invention may comprise Bacillus subtilis. For example, the at least one bacillus strain of (iv) may comprise bacillus subtilis DSMZ 32881. In an alternative embodiment, any commercially available strain of Bacillus may be used.

Advantageously, the at least one bacillus strain added produces certain essential enzymes for the biodegradation of human excreta, more specifically, promotes the production of important enzymes for the rapid biodegradation of excreta.

More advantageously, all strains of (i) to (iv) of the first and second aspects of the invention are safe to use.

Each isolated microbial strain, at least one lactic acid and/or acetic acid producing microbial strain, at least one yeast strain and at least one bacillus strain from the excreta may be cultured separately before being combined to produce a microbial flora for degrading waste. In an alternative embodiment, an isolated microbial strain from excrement, at least one lactic acid and/or acetic acid producing microbial strain, at least one yeast strain and at least one isolated bacillus strain may be combined and co-cultured to produce a microbial flora for degrading waste.

The method for preparing a product for degrading waste may further comprise immobilizing a microbial flora on the solid medium. Any suitable solid medium can be used for immobilizing the substantially pure culture, microbial flora, or mixed microbial composition. Cheap and biodegradable solid media are preferred. Examples of suitable solid media include, but are not limited to, sawdust, spent grain, or solid media from the oil palm industry, such as empty fruit clumps of oil palm.

The invention includes a product for degrading waste comprising a microbial flora or a mixed microbial composition as described herein.

The invention also includes a method of degrading waste, the method comprising: providing one of a microbial flora or a mixed microbial composition and a product for degrading waste as described herein, mixing one of a microbial flora or a mixed microbial composition and a product for degrading waste with waste; and biodegrading the waste with one of a microbial flora or a mixed microbial composition and a product for degrading the waste. The waste may be biodegraded at a temperature between about 20 degrees celsius (° c) and about 50 ℃, and more preferably, between about 30 ℃ and about 50 ℃. The waste may comprise excrement.

Having now generally described the invention, the same will be more readily understood through reference to the following examples, which are provided by way of illustration and are not intended to be limiting of the present invention.

Examples

Such as Green and Sambrook, molecular cloning: a laboratory manual, cold spring harbor laboratory, described in new york (2012), generally follows standard molecular biology techniques known in the art and not specifically described.

Example 1: screening of strains from human excrement (feces)

Screening was accomplished by two methods.

In the first method, a small amount of human excrement (feces) (10mg) was placed in a culture tube of 5ml tryptic soy broth (TSB, sigma aldrich, usa) and grown overnight at 30 ℃ and 200 rpm. The resulting culture was streaked on tryptic soy agar (TSA, sigma aldrich, usa) and grown overnight at 30 ℃. Colonies on TSA were subsequently subcultured for 16S ribosomal dna (rdna) analysis.

In the second method, 100mg of human excrement (feces) was placed in a 30ml flask containing yeast extract-peptone-glycerol (YPG) medium (10g/L yeast extract, 20ml/L glycerol, 20g/L peptone) containing 4% ethanol, and grown overnight at 30 ℃ and 200 rpm. Spreading on yeast extract-glucose-salt (YGS) agar (20g/L glucose, 20g/L yeast extract, 5g/L NaCl, 2g/L KH)₂PO₄15g agar) were previously serially diluted. Agar plates were grown overnight at 30 ℃. Single colonies on YGS agar were subsequently subcultured and used for 16S ribosomal DNA (rDNA) analysis.

All isolates from both methods were grown in their respective media and about 2ml of cell culture was collected for each strain, centrifuged and drained to obtain a cell pellet for identification. Genomic DNA of each isolate was extracted using DNeasy Blood and Tissue Kit (DNeasy Blood and Tissue Kit) (Qiagen, the Netherlands). The 16S rDNA was amplified using Phusion high fidelity DNA polymerase (Sammer Feishell technology, USA) and universal primers (Fwd 5'-AGAGTTTGATCATGGCTCAG-3' and Rev 5'-AAGGAGGTGATCCAGCCGCA-3'). PCR was performed on a G-Storm 2 thermocycler (G Storm, UK) under the following conditions: initial denaturation at 98 ℃ for 30 seconds followed by 30 cycles of denaturation at 98 ℃ for 30 seconds, annealing at 55 ℃ for 20 seconds, and extension at 72 ℃ for 45 seconds. Final extension was carried out at 72 ℃ for 10 minutes. The resulting PCR products were electrophoresed on a 1% (w/V) agarose gel at 120V for 30 minutes, gel purified using the Qiagen QIAquick gel extraction kit (Qiagen, the Netherlands) and sequenced. Homology searches were performed on the isolated 16S rDNA using nbast on NCBI website and species identification was analyzed. Finally, three biologically safe grade 1 isolates were selected for the preparation of microbial populations.

TABLE 1 selected isolates from human solid waste (faeces)

Example 2: screening of acetic acid and lactic acid producing microorganisms from commercial sources

Commercially available acetic acid and lactic acid microbial powders were purchased from Tanbao (Ali baba group, China). To isolate the acetic acid strain, a small amount of the powder (10mg) was placed in a 5ml YPG medium (10g/L yeast extract, 20g/L peptone, 20ml/L glycerol and 50ml/L ethanol) supplemented with ethanol in a culture tube and grown at 30 ℃ and 200rpm for 2-3 days. The resulting culture was streaked on solid agar of the same medium and grown at 30 ℃ for 2 days. Colonies were subsequently subcultured for 16S ribosomal DNA (rDNA) analysis. To isolate the lactic acid bacterial strains, small amounts of powder (10mg) were placed in 5ml culture tubes of MRS broth (Sigma Aldrich, USA) and grown overnight at 30 ℃ and 200 rpm. The resulting cultures were streaked on MRS agar plates and grown at 30 ℃ for 1 day. Isolated colonies of all acetic acid and lactic acid microorganisms were identified using a procedure similar to that described in example 1 above, except for yeast strains that required different primer sets (F-5665 '-CAGCAGCCGCGGTAATTCC-3' and R-12005 '-CCCGTGTTGAGTCAAATTAAGC-3'). The identification of the strains is shown in table 2.

TABLE 2 identification of acetic acid and lactic acid producing microorganisms and yeast strains

Isolate	Identification
		AA2	Saccharomyces cerevisiae
AA8	Acetobacter pasteurianus
		Lacid 3-1	Lactobacillus genus
Lacid 12	Lactococcus lactis

Example 3: development of WM8 for composting human Excreta

A new microbial flora WM8 was developed by mixing three isolates from human waste (table 1), four isolates from commercial sources and a culture of bacillus subtilis (DSMZ 32881) isolated from food waste. All strains were grown in their respective media and temperatures as listed in table 3 below. Starting cultures of all strains were prepared in 20-50 ml medium in shake flasks, and colonies on agar plates were inoculated directly into the respective media and grown overnight at 200rpm and the preferred temperature. Thereafter, the starting culture was inoculated at 5-10% (v/v) into 100ml of each medium and grown at 200rpm and a preferred temperature for 2 days. To concentrate the culture for higher cell density, the culture was transferred to a 500ml centrifuge flask and centrifuged at 10 ℃ and 8,000rpm for 5 minutes. Half of the supernatant was discarded and the remaining contents of each culture were mixed well.

Table 3: media and temperatures for culturing the respective isolates

The mixed cell culture was fixed by adding wood chips until no free water was visible. The immobilized cells (about 5kg) were stabilized at 30 ℃ for 24 hours and continuously spun for use, as shown in FIG. 1.

Example 4: composting of human waste by immobilised microbial flora

Human waste (180 g of feces and 50g of urine) was added to the immobilized microbial flora in the bio-toilet room as shown in fig. 2, and the mixture was thoroughly mixed and maintained at 20-50 ℃ by spinning. After 1 hour, human waste was almost invisible. After 24 hours, 100% of the human waste was degraded and the compost looked like dry powder fertilizer, as shown in figure 3.

Discussion of the related Art

By isolating microorganisms from human excreta and combining the isolated microbial strains with selected other microbial strains, a new microbial flora WM8 was developed. The microbial flora may be immobilized on an inexpensive solid support to increase the stability of the microbial flora for commercial use. Human excreta (feces + urine) are almost completely degraded within 24 hours of treatment at 20-50 ℃ using a fixed microbial flora.

The microbial flora may consist of seven bacterial strains and one yeast strain. These may be obtained from one or more toilet samples, food sources, or commercial sources, and may all be biologically safe grade 1 strains.

Microbial strains make composting of human waste possible without having to separate urine from feces. The microbial flora is capable of composting human waste (feces and urine) more efficiently than commercially available microbial flora, with a shorter composting time of less than 24 hours, with a lower composting temperature of 20-50 ℃, and with a higher reduction rate of human waste of more than 90%. Advantageously, this reduces composting time and energy consumption and also increases composting efficiency, thereby benefiting the environment and economy. More advantageously, the microbial flora may be used in bio-toilet applications in the absence of flush water.

While the preferred embodiments of the invention have been illustrated and described, it will be clear that the invention is not limited to the embodiments described. Numerous modifications, changes, variations, substitutions and equivalents will be apparent to those skilled in the art without departing from the scope of the invention as described in the claims.

Claims

1. A method of making a product for degrading waste comprising:

(i) providing at least one isolated microbial strain from an excreta;

(iii) providing at least one yeast strain;

(iv) providing at least one strain of Bacillus, and

(v) (iii) combining microbial strains from (i) to (iv) to produce a microbial population for degrading waste.

2. The method of claim 1, wherein the strain from (i) is selected from the group consisting of Paenibacillus, Bacillus, and Pediococcus.

3. The method of claim 2, wherein the Paenibacillus comprises Paenibacillus DSMZ 32878.

4. The method of claim 2 or 3, wherein the Bacillus comprises Bacillus DSMZ 32879.

5. A method according to any one of claims 2 to 4 wherein the Pediococcus species comprises Pediococcus lactis.

6. The method of claim 5, wherein said Pediococcus comprises Pediococcus lactis DSMZ 32880.

7. The method of claim 2, wherein the strain from (i) consists of paenibacillus DSMZ32878, bacillus DSMZ 32879 and pediococcus acidilactici DSMZ 32880.

8. A process according to any one of the preceding claims, wherein the strain from (ii) is selected from the group consisting of acetobacter, Lactobacillus (Lactobacillus sp.) and lactococcus.

9. The method of claim 8, wherein the Acetobacter comprises Acetobacter pasteurianus.

10. The method of claim 8 or 9, wherein the genus lactococcus comprises lactococcus lactis.

11. The process according to claim 8, wherein the strains from (ii) consist of Acetobacter pasteurianus, Lactobacillus and lactococcus lactis.

12. The method according to any one of the preceding claims, wherein the at least one yeast strain from (iii) comprises saccharomyces.

13. The method of claim 12, wherein the saccharomyces comprises saccharomyces cerevisiae.

14. The process according to any one of the preceding claims, wherein the at least one strain of bacillus from (iv) comprises bacillus subtilis.

15. The method of claim 14, wherein the at least one bacillus strain from (iv) comprises bacillus subtilis DSMZ 32881.

16. A method according to any one of the preceding claims, wherein each of the microbial strains in (i), (ii), (iii) and (iv) is cultured separately prior to combining to produce a microbial flora for degrading waste.

17. A method according to any one of claims 1 to 15, wherein the microbial strains of (i), (ii), (iii) and (iv) are combined and co-cultured to produce a microbial flora for degrading waste.

18. The method of any one of the preceding claims, wherein the method further comprises immobilizing the microbial population on a solid medium.

19. The method of claim 18, wherein the solid medium is sawdust, spent grain, or empty grove from oil palm.

20. A microbial flora or mixed microbial composition comprising:

(i) at least one isolated microbial strain from excrement;

(ii) at least one lactic acid and/or acetic acid producing microbial strain;

(iii) at least one yeast strain; and

(iv) at least one strain of bacillus.

21. The microbial flora or mixed microbial composition of claim 20, wherein said strain from (i) is selected from the group consisting of paenibacillus, bacillus and pediococcus.

22. The microbial flora or mixed microbial composition of claim 21, wherein said Paenibacillus comprises Paenibacillus DSMZ 32878.

23. The microbial flora or mixed microbial composition of claim 21 or 22, wherein said bacillus comprises bacillus DSMZ 32879.

24. The microbial flora or mixed microbial composition of any one of claims 21-23, wherein the Pediococcus comprises Pediococcus lactis.

25. The microbial flora or mixed microbial composition of claim 24, wherein said Pediococcus comprises Pediococcus lactis DSMZ 32880.

26. The microbial flora or mixed microbial composition of claim 21, wherein said strain from (i) consists of bacillus paenibacillus DSMZ32878, bacillus DSMZ 32879 and pediococcus acidilactici DSMZ 32880.

27. The microbial flora or mixed microbial composition of any one of claims 20-26, wherein said strain from (ii) is selected from the group consisting of Acetobacter, Lactobacillus, and lactococcus.

28. The microbial flora or mixed microbial composition of claim 27, wherein said acetobacter genus comprises acetobacter pasteurianus.

29. The microbial flora or mixed microbial composition of claim 27 or 28, wherein said lactococcus comprises lactococcus lactis.

30. The microbial flora or mixed microbial composition of claim 27, wherein said strains from (ii) consist of acetobacter pasteurianus, lactobacillus and lactococcus lactis.

31. The microbial flora or mixed microbial composition of any one of claims 20-30, wherein the at least one yeast strain from (iii) comprises saccharomyces.

32. The microbial flora or mixed microbial composition of claim 31, wherein said saccharomyces comprises saccharomyces cerevisiae.

33. The microbial flora or mixed microbial composition of any of claims 20-32, wherein at least one bacillus strain from (iv) comprises bacillus subtilis.

34. The microbial flora or mixed microbial composition of claim 33, wherein at least one bacillus strain from (iv) comprises bacillus subtilis DSMZ 32881.

35. The microbial flora or mixed microbial composition of any one of claims 20-34, immobilized with a solid medium.

36. The microbial flora or mixed microbial composition of claim 35, wherein said solid medium is sawdust, spent grain, or empty grove from oil palm.

37. A product for degrading waste comprising the microbial flora or mixed microbial composition of any one of claims 20-36.

38. A method of degrading waste comprising:

providing one of a microbial flora or a mixed microbial composition of any one of claims 20-36 and a product for degrading waste of claim 37;

mixing one of a microbial flora or a mixed microbial composition and a product for degrading waste with the waste; and

biodegrading the waste with one of a microbial flora or a mixed microbial composition and a product for degrading the waste.

39. The waste degradation method of claim 38, wherein the waste is biodegraded at a temperature between about 20 ℃ and about 50 ℃.

40. A waste degradation method as claimed in claim 38 or 39 wherein the waste comprises excrement.