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Application of volatile antibiotics and non-volatile inhibitors from muscodor spp. to control harmful microbes in human and animal wastes

a technology of volatile antibiotics and inhibitors, which is applied in the field of application of volatile antibiotics and non-volatile inhibitors of muscodor to control harmful microorganisms in human and animal waste, and can solve problems such as environmental and health hazards

Inactive Publication Date: 2006-06-15
MONTANA STATE UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0015]FIG. 4. A control experiment of the one illustrated in FIG. 3 in which M. albus was not placed on the four quarter plate and thus all bacteria grew profusely.
[0016]FIG. 5. The split plate agar assay system and the inhibition of bacterial growth from human wastes. The white growth is M. albus, and it is on both a split plate and a solid agar (PDA) plate with a streak of human solid wastes (right side of plates). The control plates have no M. albus growth, and the bacterial growth from the human excrement is enormous. In the other cases, the M. albus does inhibit bacterial growth in human wastes, but the inhibition is greater when the fungus is not on the split plates, which suggests the presence of non-volatile inhibitors.

Problems solved by technology

Unfortunately, in many cases, both environmental as well health hazards are caused when these wastes are not properly disposed (FIG. 1).
The former two bacteria are found in human wastes and are pathogenic to humans.

Method used

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  • Application of volatile antibiotics and non-volatile inhibitors from muscodor spp. to control harmful microbes in human and animal wastes
  • Application of volatile antibiotics and non-volatile inhibitors from muscodor spp. to control harmful microbes in human and animal wastes
  • Application of volatile antibiotics and non-volatile inhibitors from muscodor spp. to control harmful microbes in human and animal wastes

Examples

Experimental program
Comparison scheme
Effect test

example 1

A. Example 1

Analysis of the Volatiles of M. albus

[0026] A method was devised to analyze the gases in the air space above the M. albus mycelium growing in Petri plates. First, a “Solid Phase Micro Extraction” syringe was shown to be a convenient method for trapping the fungal volatiles. The fiber material (Supelco) was 50 / 30 divinylbenzene / carburen on polydimethylsiloxane on a stable flex fiber. The syringe was placed through a small hole drilled in the side of the Petri plate and exposed to the vapor phase for 45 minutes. The syringe was then inserted into a gas chromatograph (Hewlett Packard 5890 Series II Plus) equipped with a mass-selective detector. A 30 m×0.25 mm I.D. ZB Wax capillary column with a film thickness of 0.50 mm was used for the separation of the volatiles. The column was temperature programmed as follows: 25° C. for 2 minutes followed to 220° C. at 5° C. / minute. The carrier gas was Helium Ultra High Purity (local distributor), and the initial column head pressure ...

example 2

B. Example 2

Bioassay of M. albus against Human Pathogens

[0029] A relatively simple bioassay test system was devised that allows only for volatiles being the causative agents for any microbial inhibition. Initially, on a Petri plate with PDA, an agar strip 2.5 cm wide was completely removed from the mid-portion of it (FIG. 2). Then, M. albus was inoculated and grown on one side of the plate for varying time periods prior to testing. The test fungus or bacterium was placed onto the agar half moon strip on the opposite side of the plate. Individual fungi were inoculated on the test side of the plate on a 3 mm3 plug of agar. Bacteria and Candida albicans were simply streaked (1.5 cm long) onto the PDA on the test side of the plate. The act of removing a strip of agar from the mid-portion of the plate effectively precluded the diffusion of any inhibitory soluble compounds emanating from M. albus to the fungi or bacteria being tested (FIG. 2). The plate was wrapped with two individual pi...

example 3

C. Example 3

Direct Petri Plate Inhibition Assays

[0031] While the quadrant plate technique unequivocally demonstrated that volatiles diffusing from M. albus were inhibitory to microbes found in human wastes such as E. coli and Vibrio cholerea, evidence of other inhibitory compounds in M. albus was sought. To this end, ten-day-old cultures of M. albus on half plates, with and without the center strip removed (known as a split plate), were incubated with streaks of human solid wastes for seven days. Appropriate controls, without M. albus, were also run simultaneously. The results showed that virtually no bacterial growth occurred on the plate with M. albus, the human wastes, and no removal of the agar strip. Likewise, some inhibition occurred on the plate with M. albus, human wastes, and the agar strip removed, which suggests that the volatiles of M. albus were affecting bacterial growth but not as extensively as when direct contact of the wastes and the fungal culture existed. As exp...

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Abstract

This invention relates to the discovery of a novel endophytic fungus, Muscodor albus, which produces volatile antibiotics that have been shown to be effective in the treatment of human and animal waste products. Through experimentation, this invention confirms that Muscodor albus can be used in disposable bags in connection with portable toilets to solve the problem of degradation of waste products in situations where humans are removed from sanitary facilities. This invention includes the discovery of non-volatile inhibitors that are also produced by Muscodor albus and that are similarly effective in treating human and animal wastes. This invention covers a unique and novel method for preparing Muscodor albus for commercial use in the treatment of human and animal wastes, which involves infesting barley seeds with Muscodor albus and then storing them under conditions that will allow the fungus to remain viable for up to nine months.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a divisional application of U.S. application Ser. No. 10 / 408,209, filed Apr. 4, 2003. The latter application in turn claims the benefit under 35 U.S.C. § 120 and is a continuation-in-part of U.S. Nonprovisional Application No. 10 / 121,740, filed Apr. 11, 2002, which in turn claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 60 / 283,902, filed Apr. 16, 2001 and of U.S. Provisional Application No. 60 / 363,072, filed Mar. 11, 2002. The contents of these applications are hereby incorporated by reference into the present disclosure.BACKGROUND OF THE INVENTION [0002] Throughout this application, various articles and books are referenced by authorship and date. The full bibliographic citation for each publication can be found at the end of the specification, immediately preceding the claims. In addition, certain terms used in this application are defined in the definitions section immediately follo...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61L9/015A61K35/74A01N63/30C12N1/14
CPCA01N63/02C12N1/14A01N63/04A01N63/30
Inventor STROBEL, GARY ALLANEZRA, DAVID
Owner MONTANA STATE UNIVERSITY
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