WO2011042684A2

WO2011042684A2 - Antimicrobial composition and method of controlling contamination or infections using said composition

Info

Publication number: WO2011042684A2
Application number: PCT/GB2010/001837
Authority: WO
Inventors: Alyson Bexfield; Alison Elisabeth Bond; Edward Dudley; Peter Russel Newton; Yamni Nigam; Norman Arthur Ratcliffe
Original assignee: Uws Ventures Limited
Priority date: 2009-10-08
Filing date: 2010-10-01
Publication date: 2011-04-14
Also published as: WO2011042684A3; GB0917554D0; GB2474251A

Abstract

The invention relates to an antimicrobial composition having the formula C₅H₁₁O₂N, and salts or esters thereof, which is obtained, in particular from extracts of fly larvae.

Description

Antimicrobial Composition and Method of Controlling Contamination or infections using said composition

The present invention relates to an antimicrobial composition and to formulations using said composition in order to control contamination and/or infections. Further, the invention relates to a method of treatment using the antimicrobial compositions of the invention. In particular, the invention relates to compositions used as anti-fungal agents.

Antimicrobial compositions are known and can be used to treat a wide range of microorganisms including fungi, yeasts and protozoa. Further antimicrobials can be useful in treating contaminations involving a number of different species of microorganisms, such as bacteria in combination with a fungal infection. In particular some illnesses, such as those involving the respiratory tract and the lungs, can result from infections involving both bacteria and fungi making such infections difficult to treat because the treatment needs to be effective against very different types of organisms and so several different types of treatments are given simultaneously to control the overall infection caused by the different types of organisms.

Much general research has been done in the field of antimicrobials and fungal/yeast infections. It is known that agricultural workers can be at risk from the airborne spores of fungi which often form the fungal flora in cereal crops. A particular problem results from contaminations by my- cotoxins and fusariotoxins produced by fungi belonging to the genus Fusarium. The spores from dust released from the surface of grain can lodge in the lungs and cause persistent and long term lung infections for agricultural workers. Contamination of grain can be particularly high if grain has been stored when damp, so allowing fungi to grow. Also, yeast, which are classified as fungi are naturally on the skin and in most circumstances, cause no harm but if individuals have a reduced immune system, for example if they are ill with conditions such as cancer or AIDS, then what would be a normal body flora organism, can cause severe and even life threatening infections of the skin and organs. In cancer patients, infections with organisms from the yeast species Candida can be very distressing because of the pain and discomfort caused.

A more recent problem with many antibiotics or healing agents is that they are often only partially effective. In particular in the case of antibiotics, as a result of widespread and indiscrimi- nate use, microbes have become more resistant to the microbial effects of such antibiotics and a prime example of such resistance can be seen with the so called "superbugs" such as methicillin resistant Staphylococcus Aureus (MRSA). Increased resistance is also now being observed in fungi and yeasts. A consequence of increased resistance by microorganisms to antibiotics and anti-fungals is that treatments and decontamination procedures become more costly as more and more sophisticated agents are needed to kill the microorganisms.

An additional problem in the case of fungi and yeasts, is that because they are eukaryotic organisms, as are humans and animals, there can be complications in treatments because of side- effects due to a composition being used to treat similar types of cells. Further, fungi and yeast have several stages in their life cycle, for example, spores, haploids, zygotes, diploid colonies and then on to spores, which makes fungal infections difficult to treat because a reagent is needed that is effective against the fungi/yeasts at all stages in the life cycle because if only a proportion of the organisms are killed, then the remaining organisms can establish new colonies and infections are repeated. In addition, because the infections are due to an existing colony, there is an increased chance of resistance to control reagents by organisms from the original colony.

Work has been carried out in order to isolate biologically active substances to treat bacterial infections using secretions from larvae from green-bottle flies. The application of sterile larvae from Lucilia sericata to an infected wound results in the removal of necrotic and infected tissue. The application of sterile whole fly larvae to a wound is generally known as maggot therapy. It is recognised that there is a degree of mechanical irrigation of the wound by physical digestion of tissue by larvae. However, it is also noted that there is antimicrobial action caused by secretions from the maggots and work has been done by The University of Nottingham and published in PCT patent application number WO 03/075654, where they have taken complete secretions/excretions from Lucilia sericata treat wounds and used these secretions with known antibiotics to inhibit the growth of Gram -i-ve and Gram -ve bacteria. This publication clearly states though that the secretions themselves have no antibiotic effect, it is the combined effect of the secretions with a known antibiotic that inhibits bacterial growth. In a series of German applications, DE 10149143A 1 , DE 101383O3A 1 and DE 19901 134, again work has been done on enzymes or enzymatic extracts from whole body homogenates of the larvae or pupae of flies such as Sarcophaga or L cilia. These extracts are used as general antimicrobials and they contain a mixture of components, but there is no discussion of specific purified components from the secretions.

Researchers at the University of Swansea recognized the need for isolating a specific compound that would be an effective antimicrobial, especially as administering complete exudates may be unacceptable to patients and medical personnel alike. In addition, by having a specific isolate, the treatment program for an individual can be more precisely controlled because using a specific isolate allows for providing a correct and efficient dosage as cost effectively as possible, rather than having a secretion where it is not known what components are effective. A specific composition from Lucilia sericata has been isolated and described in WO 2008/087416 and this composition has been found to be particularly effective against MRSA bacteria.

This known work has all been directed to looking at prohibiting the growth of bacteria. It is known that fungal infections can be particularly problematical to treat because there are multiple stages in the life cycle of the fungi/yeasts. As a result of this it has been very difficult to isolate compounds that will be effective in treatments. Further, there is the added problem that fungi/yeasts are eukaryotes and so it is necessary to find a compound that will be effective against the fungi/yeasts, without also being harmful to the cells of the organism that is being treated.

The present invention seeks to overcome the problems of the prior art by providing a defined antimicrobial agent which in particular has effective anti-fungal activity. Further , the antimicrobial is effective against fungi/yeasts at various stages in their life cycle, without being harmful to the organisms being treated which has the particular advantage of reducing the chances of the fungi/yeasts becoming resistant to treatment because all of the colony of fungi/yeasts are killed.

It is to be understood that the term antimicrobial covers compounds and combinations of compounds that have bacteriostatic, bacteriocidal, fungistatic and fungicidal properties and which can be administered to a human or an animal to inhibit infection. It is even envisaged that the anti-fungal can be used to treat plants, which can be particularly prone to fungal infections.

Although the antimicrobial is particularly applicable to wounds it may be used to treat specific infections for example fungal infections of the skin surface or infections where there is a combination of fungi and or yeasts as well as bacteria. The antimicrobial may be administered topically or systemically for the treatment of infection. The antimicrobial may also be used in other applications such as, but not limited to use as a disinfectant, as an antiseptic, as a surface cleaner or even as a preservative in compositions or final products produced in the food industry or in the cosmetics industry.

According to a first aspect of the invention there is provided an antimicrobial composition having the formula C5H 1 1 O2N, and salts or esters thereof.

Preferably the antimicrobial composition is a composition that is effective against fungi or yeasts.

It is preferred that the antimicrobial composition is isolated from excretions/secretions of fly larvae, although it is envisaged that it may be isolated from other sources such as from vertebrates or invertebrates.

Preferably the antimicrobial is obtained from fly larvae of the species L cilia sericata.

In a further aspect of the invention, there is provided a pharmaceutical composition including an effective amount of an antimicrobial composition of the formula C5H 1 i0₂N and salts or esters thereof.

The pharmaceutical composition preferably comprises a suitable dosage of the antimicrobial composition together with a pharmaceutically suitable and physiologically tolerated carrier. Where appropriate other suitable substances may be included in the pharmaceutical composition such as other active substances, additives or excipients. The pharmaceuticals of the invention may be applied topically but they may also be applied in- tracorporally, for example intravenously or by implants in the body or by way of transdermal application, for example skin patches. A further way of administering the pharmaceutical composition may be ingestion orally or by using suppositories.

Suitable pharmaceutical compositions for topical use on the skin are preferably in the form of a solution, a suspension, a dusting powder, liposomal formulations, gels, lotions, pastes, sprays or aerosols. Carriers that can be used include polyethylene glycols, alcohols and combinations of two or more of these substances and the list is not to be regarded as restrictive. Sprays and dusting powders are particularly effective in treating areas of the body where it may be hard to reach, for example between the toes as a common ailment for feet is the fungal infection by tinea pedis, which causes athlete's foot. In the case of infections that cause conditions affecting the scalp, for example ring worm or dandruff, the route for administering the antimicrobial is via a shampoo or hair wash.

The antimicrobial composition of the invention preferably is present in a concentration from 0.1 % by weight to 100% by weight of the composition, for example 50-90%, or 40-80%, 30- 70% or from 1.0% by weight to 60% by weight, depending on the modes of extraction of the antimicrobial composition.

In the case of fungal/yeast infections it is particularly useful if the compositions can be given transdermally because fungi/yeast cause many skin infections. A particularly useful way of administering the effective composition is by using dressings which contain the antimicrobial composition. Individual plasters may be used to provide long-term close contact of the composition with the skin whilst providing some protection for open wounds. A suitable active substance concentration is from 0.01 % by weight to 75% by weight, preferably 1 % by weight to 70% by weight.

The antimicrobial of the invention can also be applied to a wound though coverings made of gauze, of alginates or hydrocolloid materials, foams or silicones coverings, or combinations of the materials listed. The coverings may be coated, impregnated or treated with the composition or there may be a reservoir in the dressing holding the composition so it can leach out through the dressing and onto the skin surface and into a wound over a period of time.

The composition may also be delivered as an oral pharmaceutical such as a tablet but it may also be provided as a solid pharmaceutical.

Suitable solid pharmaceutical are, for example, granules, powders, products with protracted release of active substances where conventional excipients or carriers are used. Excipients which are frequently used are magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, milk protein, gelatine, starch, cellulose and derivatives thereof, polyethylene glycol and solvents, such as for example sterile water, monohydric or polyhydric alcohols such as glycerol.

According to a further aspect of the invention the pharmaceutical composition is arranged to be administered intravenously.

The carrier composition may be any pharmaceutically acceptable carrier such as a solvent for antibiotics, chelating agents and pH buffering agents that will allow a therapeutic composition to be administered directly to the skin, wound or mucosa or systemically by, for example but not limited to, oral or intravenous administration. The carrier may also allow the use of the antimicrobial for other purposes, for example but not limited to, antiseptics, disinfectants and preservatives. Examples of carriers that can be used, which is not a limiting list, include water, saline, physiological saline, ointments, creams, oil in water emulsions, gels, solvents or combinations of solvents.

The treatment preparation also includes a pH buffering agent. The term pH buffering agent includes, for example, pharmaceutically acceptable organic or inorganic compounds or combinations of compounds that will maintain the pH of the treatment preparation within desired pH limits. It is envisaged that the treatment preparation further includes a surfactant to allow for adequate mixing of the antimicrobial in a carrier and this is particularly applicable where the antimicrobial is provided in a shampoo or body wash. Known surfactants, such as detergents, or anionic or ionic surfactant or a mixture thereof may be used. Surfactants are included to alter the surface tension of aqueous solutions containing the antimicrobial composition and therefore provide a treatment preparation that can be easily taken into the body. It is envisaged that the carrier may include one or more of a pH buffering agent, a tissue growth promoting agent, an antiinflammatory or a pain ki ller and a surfactant or any such agents which allow administration via, for example, oral, parenteral or intravenous methods.

The treatment preparation may also include a wound healing promoter such as Vitamin E which can assist in the skin around a wound healing by promoting tissue growth .

In addition the treatment preparation may include, such as, one or more anti-inflammatory agents for example a corticosteroid or agents that can reduce pain, such as morphine based substances. These inclusions are particularly useful if the patient is feeling uncomfortable because the wound damage is deep seated.

In addition to treatment of the body, the antimicrobial composition may be incorporated in a formulation to treat food to reduce the risk of microbial contamination.

Further the antimicrobial may be incorporated in a disinfectant to be used as a general surface cleaner, for example, for floor coverings such as carpets or for hard surfaces in bathrooms, kitchens and on floors.

The antimicrobial composition may also be used as a general antiseptic, which can be used in a range of environments, not only in the house, but in hospitals, workplaces, in areas where other sterile environments are required. The antimicrobial may be incorporated in for example antiseptic wipes or in misters or it can be used as a liquid or gel in the form of a surface cleaner. Further, the composition claimed when used as a medicament may be provided in the form of a salt, an ester or pro drug derivative.

According to a further aspect there is provided a method of treating a wound infection, said method comprising contacting a wound with a therapeutic amount of an antimicrobial composition of the empirical formula C5H1 |0₂N or and salts or esters thereof.

Furthermore, infection may be treated by, but not limited to, oral, intravenous, or parenteral administration of the antimicrobial composition.

Preferably, the antimicrobial composition is an isolate from the secretions/excretions of fly larvae.

It is envisaged that preferably the species of fly larvae is L cilia sericata.

Preferably, the antimicrobial composition is included in a pharmaceutically acceptable carrier.

It is envisaged that the carrier may include one or more of a pH buffering agent, a tissue growth promoting agent and a surfactant.

Treatment is preferably oral, or failing that, parenteral.

According to yet further aspect of the invention, there is provided a wound dressing including an antimicrobial composition having the empirical formula of C5H 1 |0₂N and salts or esters thereof.

Preferably the wound dressing includes a treatment preparation including an antimicrobial composition of the empirical formula C₅H n0₂N and a carrier composition for the antimicrobial composition.

It is envisaged that the carrier may include one or more of a pH buffering agent, a tissue growth promoting agent, an anti-inflammatory or a pain killer and a surfactant. Although separate aspects or embodiments of the invention have been described, the invention is intended to also cover combinations of those aspects or embodiments.

The following examples are provided to illustrate preferred embodiments of the invention and uses of the present invention are provided by way of example only and should not be construed to limit the scope of the invention.

Example 1

METHODS AND MATERIALS

third instar larvae of Lucilia sehcat were obtained from a supplier of fly larvae.

Microorganisms used in assays

The yeast, Candida albicans ATCC 2091 , was purchased from NCIMB (National Collection of Industrial, Marine and Food Bacteria), Aberdeen, UK. C. albicans 2851 8 and C. kruseii 6255 were a gift from Prof. Steve Kelly, Institute of Life Science, Swansea University, UK. Yeast cells were grown on Sabouraud dextrose agar (SDA; Fisher Scientific Ltd.) for 48 h at 37°C. A colony was then picked and suspended in 0.03% Tween 80 (Sigma Aldrich, Dorset, UK) at a concentration of 1 x 10⁶ yeast cells/ml. The filamentous fungi, Metharizium anisopliae 4556 and Beauvaria bassiana, were provided by Dr T. Butt, Swansea University. Paecilomyces lilacinus was purchased from the National Collection of Pathogenic Fungi (NCPF), UK. Filamentous fungi were grown on SDA at 25°C for approximately 20 days or until the cultures sporulated. Fungal spores were suspended in 0.03% Tween 80 at a concentration of 1 x 10⁷ spores/ml and stored at 4°C for up to one week until required. Staphylococcus aureus 9518, was purchased from the NCIMB. E. coli K 12 was purchased from the Coli Genetic Stock Center (CGSC), New Haven, USA. Bacteria were grown for 17 h in 10 ml of tryptic soy broth (TSB, Difco, Becton Dickinson UK Ltd., Oxford, UK). Collection of Larval excretions/secretions

Native excretions/secretions (nES) were collected by incubating third-instar larvae in a small quantity (200 μΙ g^{' 1} ) of sterile Milli-Q water (Millipore UK Ltd., Hertfordshire, UK) for 1 h at 30°C in darkness, following which ES (excretion/secretion) was siphoned from the containers, centrifuged at 10,000 x g for 5 min and the supernatant retained for testing. Although in this case third instar larvae were used, it is possible to use first and/or second-instar larvae but third instar- larvae yielded increased sample volume and anti-fungal activity.

Purification of the Supernatant

Pooled nES ( 10 ml) was sequentially fractionated under pressure from an oxygen free nitrogen cylinder at 40 psi. The secretions were filtered through a 10 kDa molecular weight cut-off (MWCO) membrane then a 500 Da MWCO membrane in an Amicon stirred ultrafiltration cell (Millipore UK Ltd) (Bexfield et ai., 2004). The Amicon filters generated fractions from the excretions/secretions of different molecular weights - (Bexfield, A et al, Microbes and Infection 6, 1297- 1304, 2004).

The less than 500 Da fraction was shown to be active and taken for further fractionation. Separation of the Fractions

HPLC separation was performed using a Vision Workstation (Applied Biosystems, Warrington, UK) equipped with a quaternary pump and an AFC 2000 fraction collector. Samples were separated on a BioBasic SEC-60 column (300 x 7.8 mm i.d.; Thermo Electron Corporation, Hemel Hempstead, UK). The mobile phase consisted of Milli-Q water with a flow rate of 0.5 mlJmin. The column was run at room temperature (-21 °C) and fractions collected every minute whilst UV absorbance was measured at 214 nm.

Two separations were performed: a low molecular weight size exclusion separation (Biobasic SEC6000 column, Waters) and a C I column (ThermoElectron). They were both performed on a Applied Biosystems Vision HPLC workstation with BioCAD automated autosampler/fraction collecter. 50 μ] of a ten fold concentrated <500Da sample was injected and 10 runs completed and complimentary fraction pooled and concentrated prior to anti-fungal analysis.

The BioBasic column was eluted with 100% water at a flow rate of 0.5ml/min and 1 minute fractions collected. The C l column was eluted at 0.2 ml/min with an increasing gradient of methanol in water (0- 100% methanol over 30 minutes after a 15 minute period of 100% water) and 1 minute fractions collected. Fractions 1 3- 15 from the SEC column were active and fraction 4 from the C l column was active.

The active, and test inactive, fractions were analyzed by electro-spray mass spectrometry by infusion at 4 μΐ/min after dilution to a final mobile phase of 50/50 methanol/water with 0.1 % formic acid. The only ion present in all active fractions was at m/z 1 18 (Figure l a).

Further filtration isolated the anti-fungal activity to the less then 100 Da fraction. This fraction was similarly infused and analyzed by electro-spray mass spectrometry and again the m/z 1 1 8 ion was shown to be the predominant of the 6 main ions originating from the sample (Figure l b).

Assays

Following filter sterilization (0.2 μιη), ES <500 was tested for anti-fungal activity using the anti- Candida assay.

1. Anti-Candida assay

Candida were grown overnight at 37°C on Sabouraud Dextrose Agar (SDA; Oxoid Ltd, Basingstoke, UK). A single colony was picked with a sterile loop, suspended in 300 μΐ of 0.03 % Tween 80 and diluted to 1 x 10⁶ yeast ml^{" 1}. Eighteen μΐ of ES <500 were mixed with 2 μΐ of 10- fold concentrated Sabouraud Dextrose Broth (SDB ; Oxoid Ltd, Basingstoke, UK) prepared in 200 mM HEPES, pH 8.5, to give a final concentration of 1 -fold SDB and 20 m HEPES . Samples were then mixed with 2 μΐ of yeast suspension and incubated at 37°C for 24 h. In controls, ES <500 was replaced with sterile Milli-Q water. Following incubation, samples were diluted 1 : 1000 in PBS, 5 μΐ aliquots plated onto SDA, and plates incubated for 48 h at 37°C. 2. Spore Inhibition assay

Test samples (20 μΙ) were mixed with 4 μΐ of 200 mM MOPS, pH 8.5 and 4 μΐ of fungal spore suspension in 1.5 ml micro-centrifuge tubes. Tubes were incubated at 25°C for 24 h, after which 5 μΐ aliquots of sample were transferred onto microscope slides supporting 200 μΐ of solidified SDA, in triplicate. Slides were incubated in sterile Petri dishes at 25°C for 1 8 h. Following incubation, a clean glass cover slip was placed over the area of inoculation and the spores viewed through a light microscope to assess the frequency of spore germination. In controls, test samples were replaced with sterile Milli-Q water.

A potent, anti fungal factor has been identified in whole externalized, excretions/secretions of Lucilia sericata, the medicinal maggot. The factor is highly active when incubated with several fungal species. It completely inhibits the fungal growth of Candida albicans (Figure 2 and 3). Similar results are seen with Candida kruseii. The anti-fungal factor also drastically inhibits spore germination of Beauvaria Basseria (Bb l 3), Metarhizium anisophiae (V245) and Paecilo- myces lilacarius (Figure 4).

Thermal Stability of the Compound

To test the thermal stability of the anti-fungal compound, ES <500 was subjected to two treatments for heat and freeze-thaw stability, each repeated three times. For heat stability, 40 μΐ aliquots of ES <500 were placed in sealed micro-centrifuge tubes, incubated in a boiling water bath for 15, 30 or 60 min, and then samples were briefly cooled on ice and centrifuged at 10,000 x g for 5 min to pellet the denatured proteins. Samples were also tested for their stability at room temperature for extended periods. Aliquots of 40 μΙ were placed into sealed 500 μΐ microcentrifuge tubes and left at room temperature for 24 hours, 7 days and 14 days. Duplicate controls remained at -80°C for the duration of the experiment. To determine freeze-thaw stability, 40 μΐ aliquots of ES <500 were cycled from -80°C to 37°C ten times, allowing for freezing and thawing of the samples.

The anti-fungal activity detected is thermally stable, retaining activity fol lowing ten freeze-thaw cycles. Anti-fungal activity is also retained following boiling at 100°C for 60 minutes. Preliminary purification (size exclusion), of the anti fungal factor has resulted in the isolation of the anti- fungal activity to a <500 Da fraction and its separation away from an antibiotic entity (Seraticin®) also present in the secretions. We have further narrowed the size of this anti-fungal factor to a size significantly less than 500 Da, by passing it through a 100 Da cut off filter (Nominal).

In conclusion, a potent, anti fungal factor has been identified in whole externalized, excretions/secretions of L cilia sericata, the medicinal maggot. The factor is highly active when incubated with several fungal species. It completely inhibits the fungal growth of Candida albicans similar results are seen with Candida b-useii. The anti-fungal factor also drastically inhibits spore germination of and Beauvaria Basseria (Bbl 3), Melarhizium anisophiae (V245) and Pae- cilomyces lilacarius.

Aspects of the invention are now described with reference to the following figures, which are provided by way of example only. The figures show specific characteristics of the composition and that it has an ability to act over a wide range of microbes, including fungi/yeasts.

Figure la

The mass spectrometry analysis of Figure 1 a demonstrates that the compound of the invention has an empirical formula of the m/z 1 18 ion as being C₅Hi₂0₂N, hence the neutral compound would have an empirical formula of C₅H] i0₂N.

Figure lb.

In Figure 1 , the m/z 1 18 ion was fragmented to give fragment ions at m/z 100, 72 and 59. These represent a loss of water, HCOOH and approximately half the compound during fragmentation. The composition of the invention hence can be identified by the mass spectrometry graph as shown. Figure 2

The graph of Figure 2 shows that there is inhibition of the growth of Candida albicans in the presence of Native Excretions/secretions (NES) from Lucilia sericata.

Figure 3

This Figure shows the rrowth of Candida albicans in the presence of <500 Da fraction from Lucilia sericata. This graph shows that the isolated low molecular weight compound of the invention is also effective in controlling the growth of colonies when compared the control containing water.

Figure 4

Percentage spore germination of different species of filamentous fungi in the presence of

<500 Da fraction from Lucilia sericata. The graph shows the isolated compound of the present invention is highly active when incubated with several fungal species. As well as inhibiting the growth of Candida species as shown in Figures 3, similar results are seen with the inhibition of spore germination of (from left to right) Beauvaria basseria (Bbl 3), Metarhizium anisophiae (V245) and Paecilomyces lilacarius using the <500 Da fraction.

As can be seen from the Figures, the compound of the invention appears to have good efficacy and also is effective against a wide range of organisms as well as against colonies and spores of fungi, so making it a particularly effective antimicrobial. Further, the invention covers not only individual embodiments as described but combinations of the embodiments as well. It is to be understood that modifications and variations of the present invention will be apparent to those skilled in the art and it is intended that all such modifications will be included within the scope of the present invention.

Claims

1 . An antimicrobial composition having the formula C5H 1 1 O2 and salts, esters or prodrug derivatives thereof.

2. An antimicrobial composition according to claim 1 , which is an anti-fungal composition.

3. An antimicrobial composition according to claim 1 or claim 2 which is an isolate from invertebrates.

4. An antimicrobial composition according to claim 3 where the isolate is from the excretions/secretions of fly larvae.

5. An antimicrobial composition according to claim 4, wherein the fly larvae are of the species Lucilia sericata.

6. A treatment preparation including an antimicrobial composition according to any preceding claim.

7. A treatment preparation according to claim 6, including carrier composition for the antimicrobial composition.

8. A treatment preparation according to claim 7, wherein the carrier is selected from one or more of the following: water, saline, physiological saline, ointments, creams, oil in water emulsions, gels, solvents or combinations of solvents.

9. A treatment preparation according to claim 8, further including one or more of the following: a chelating agent, a pH buffering agent or a surfactant. l u. A treatment preparation according to cl ^n 8 or claim 9 including one or more wound healing promoters or one or more anti-inflammatory agents or pain reducing agents.

1 1 . A treatment preparation according to claim 10, wherein the wound healing promoter is vitamin E.

12. A wound dressing including an antimicrobial composition having the formula C5H 1 1 O2 and salts or esters thereof.

13. A wound dressing according to claim 1 2, to treat fungal/yeast infections.

14. An antiseptic, disinfectant or surface cleaner composition including an antimicrobial composition having the formula C₅H| i0₂N, and salts or esters thereof.

1 5. An antiseptic, disinfectant or surface cleaner composition according to claim 14 used to treat contamination with fungi/yeasts.

16. A plant treatment agent including an antimicrobial composition having the formula

C₅H 1 10₂N and salts or esters thereof.

1 7. A plant treatment agent according to claim 16 used as an anti-fungal anti-yeast treatment agent.

1 8. A method of treating an infection, said method including contacting an infected area with a therapeutic amount of an antimicrobial composition of the formula C5H 1 1 O2N and salts or esters thereof, or via systemic administration of said antimicrobial composition.

19. A method according to claim 18, wherein the antimicrobial is included in a pharmaceutically acceptable carrier.

20. A method according to claim 19, wherein the carrier includes one or more of a pH buffering agent, a tissue growth promoting agent, an anti-inflammatory or a pain killer.