US 2011 0212187A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0212.187 A1 Rabasse et al. (43) Pub. Date: Sep. 1, 2011

(54) ANTMICROBAL COMPOSITION Publication Classification (51) Int. Cl. (75) Inventors: Jean-Michel Rabasse, Paris (FR): AOIN 59/4 (2006.01) Johan De Saegher, Destelbergen A633/40 (2006.01) (BE); Marc Demuynck, AOIN 59/00 (2006.01) AOIN 59/06 (2006.01) Wondelgem (BE); Peter Roose, AOIP I/00 (2006.01) Sint-Martens-Latem (BE) AOIP3/00 (2006.01) AOIP5/00 (2006.01) (73) Assignee: TAMINCO, Gent (BE) AOIP 7/00 (2006.01) A6IP3L/00 (2006.01) B82Y5/00 (2011.01) (21) Appl. No.: 13/063,371 (52) U.S. Cl...... 424/615; 424/616; 424/613;977/773 (57) ABSTRACT (22) PCT Fled: Sep. 1, 2009 The invention relates to new antimicrobial compositions comprising a combination of at least one organic and/or (86) PCT NO.: PCT/EP09/61269 inorganic peroxide, —at least one source, and—at least one nitrogen containing compound which is a non glucogenic S371 (c)(1), compatible solute, selected from taurine, choline and choline (2), (4) Date: May 20, 2011 derivatives, trimethylamine-oxide (TMAO), ectoine and hydroxyectoine, the N-methylated aminoacids glycine betaine, dimethylglycine, sarcosine, carnitine, N-methylala (30) Foreign Application Priority Data nine, trimethylamino-butyric acid, butyrobetaine and proline betaine, and the poly amino hydrocarbon compounds Sep. 17, 2008 (EP) ...... O8164494.O putrescine, cadaverine, spermine and spermidine. Patent Application Publication Sep. 1, 2011 Sheet 1 of 3 US 2011/0212187 A1

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Patent Application Publication Sep. 1, 2011 Sheet 3 of 3 US 2011/0212187 A1

US 2011/0212 187 A1 Sep. 1, 2011

ANTMICROBAL COMPOSITION pathogenic bacteria and fungi, biofilm formation in piping systems, on implants, catheters, rubbers, etc. . . . . 0007 Nevertheless, silver resistant strains of microbes are 0001. The invention relates to the field of antimicrobial evolving. It is therefore interesting combining silver com compositions, also referred to as antiseptics or disinfectants. pounds with a peroxide of a combination of peroxides to 0002 Ideally, an antiseptic or disinfectant should show a prevent resistant formation. The main advantage for using broad antimicrobial spectrum with a potent germicidal activ silver compounds is the persistent antimicrobial activity over ity and a rapid activity onset combined with a long lasting a wide pH range. effect. Such formulations should not be toxic to the host 0008. The combination of hydrogen peroxide and silver tissues and should not interfere with a healing or recovery nitrate results in a synergy with a higher antimicrobial activity effect in the infected host. at a broader pH range compared to solely hydrogen peroxide. 0003 Studies with different biocides including silver Not only hydrogen peroxide but also other organic or inor compounds concluded that ionic silver kills a broad range of ganic peroxides and even mixtures of different peroxides may microbes at lower concentrations. Silver compounds are be combined with silver compounds. The resulting anti-mi therefore used in clinical practices. In other studies the com crobial activity is determined and different combinations are bined assessments of cellular cytotoxicity and microbial evaluated for the antimicrobial activity against different activity showed that the silver compounds damage host cells microbes in vitro. The silver-peroxide combination is gener more than they do to the test organisms. This toxic activity ally accepted as an ecologically friendly system for disinfec however depended strongly on the used medium. It is never tion or decontamination and many researchers suggest using theless important to reduce the stress of biocides on host these formulations as a realistic alternative for chlorine con tissues. Silver combined with peroxides should normally taining antimicrobials. Silver compounds are very reactive induce oxidative stress on the host. Therefore, it is important and are finally precipitated as metallic silver, or to introduce a stress reduction compound which does not alter silver salts. Mostly, silver is formed due to the omni the biocidal activity. Different silver salts, silver nanopar presence of chloride in the environment and all kinds of ticles and silver compound containing devices have shown process water and the extreme low of this salt. broad spectrum antimicrobial activity. Silver salts also show Silver salts also show antimicrobial activity in the form of interesting biological effects different from the antimicrobial nanoparticles. effect on host tissues. It was also demonstrated that silver 0009 Peroxide and silver compounds primarily act on the nitrate promoted plant regeneration from mature embryos in microbial cytoplasmic membrane and hereby provoke oxida different wheat cultivars which confirmed the effect on tive stress. Enzymes are denatured and the membrane shows Somatic embryogenesis and plant regeneration in a number of leakages with the release of cytoplasmic ingredients resulting dicotyledonous species of Brassica spp., pomegranate, in the death of the microbes. When such combinations are cucumber, rice, maize, pear, millet and barley. Before, it was applied on “biological Surfaces' (plant, leaves, roots, skin of demonstrated that promoted shoot regeneration animals or humans, fruits, vegetables, seeds, meet or fish from callus cultures derived from immature embryos and that products) oxidative stress occurs resulting in free radical ini it inhibits the physiological action of ethylene by competing tiated chain reactions causing deterioration of the biological for its binding site, rather than effecting ethylene synthesis. material. 0004 Silver oxide has been incorporated in nanocompos 0010 Against this complex background it has now been ites showing antibacterial activity. Different silver oxides found Surprisingly that a new antimicrobial composition can carriers such as chitosan, alginates, proteins, polysaccha be proposed resulting in a higher biocidal activity at a broader rides, gums, celluloses and others have been used as reducing pH range, based on the Synergistic activity of a peroxide stabilizing agents. The mechanisms of the antimicrobial linked to a silver compound, where this synergistic effect is action of silver ions is supposed to be linked to the action on further supplemented with well defined compounds belong Sulfhydryl groups. Addition of cysteine, or other compounds ing to the class of nitrogen containing molecules and of So with thiol groups neutralizes the silver ions action, but there is called “compatible solutes'. Surprisingly, this combination also an action on the hydrogen bounding and some essential has been found to result not only in an increased activity on enzymes causing the release of potassium ions from the bac bacteria, probably due to a normalization effect of the cyto teria. The bacterial cytoplasmic membrane has been reported plasmatic turgor, but also in a better protection of host cells to be the most essential target for silver. Silver ions have also and host tissues against oxidative stress resulting from the use been deposited in the vacuole and cell wall as granules, inhib of peroxides and from other sources of biotic and abiotic iting cell division and provoking cell damage during the StreSS. growth process. There is also supposed to exist an additional 0011 Antimicrobial compositions comprising a peroxide interaction with the DNA bases, inhibiting cell division and and a source of silver together with various additives have interfering with proton transport. been reported in the prior art. Thus, WO 00/62618 A discloses 0005. A synergistic effect between silver and copper ions in example II the use of leucine together with a peroxide and was detected. Copper ions are omnipresent in different types a silver Source, to prepare a product with germicidal potency of water. Silver nitrate is also used against warts. for aqueous systems. Leucine (a non-methylated nitrogen 0006 Silver compounds have been used for thousand of containing compound) is known as a proteogenic compound years. They showed good results and beneficial effect in many but does not act as a compatible solute nor as an osmolite (the diseases as hygienic compound, disinfectant, as compound to leucine additive in fact reacts with the silver and cannot improve would healing, diabetic ulcers, etc. . . . . The activity perform any osmolytic or other activity). GB 2 189394 A is most interesting to control microbial growth on living tis discloses in example 2 the use of Sodium or potassium ben sues and on food and feed products. The addition of silver Zoate together with a peroxide and a silver Source, to prepare compounds to water controls algae growth, development of a concentrate well Suited for disinfecting Swimming pools. US 2011/0212 187 A1 Sep. 1, 2011

Sodium or potassium benzoate is a biocide under acidic con invention. The N-methylated aminoacid, betaine type deriva ditions, not a compatible solute (osmolyte). tives, according to the invention act as compatible solutes and 0012 WO 2006/070953 A discloses a liquid purifying not as Surfactants. agent comprising silver nitrate, sodiumperoxide and aborax/ 0031. According to the invention, the silver source is pref sodium carbonate mixture. Borax is a well known biocide and erably selected from metallic silver and silver compounds. not a compatible solute. 0032. The non glucogenic compatible solute of the anti 0013 The present invention provides a new anti-microbial microbial compositions according to the invention should not composition comprising a combination of neutralize the activity of the silver compound and the con 0014 at least one organic and/or inorganic peroxide, centration should be limited and linked to the silver concen 0015 at least one silver source, and tration. 0016 at least one specific nitrogen containing com 0033. The expression “compatible solute” as used in this pound which is a non glucogenic compatible solute and context refers to its most broadly accepted sense of organic which is selected from taurine, N-methylated aminoac compounds which serve as cytoplasmic solutes to balance ids, N-methylated aminoalcohols, N-methylated amine water relations for cells growing in environments of high salt oxides, N-methylated amidines and poly amino hydro or Sugar carbon compounds, and mixtures of any of these. 0034 (or, as specifically worded in an article of Pedro 0017. According to an essential feature of the invention, Lamosa et al. (in Appl. Environ. Microbial. 1998 Octo the N-methylated aminoacids referred to above are preferably ber: 64(10): 3591-3598) “low-molecular-weight selected from glycine betaine (trimethyl glycine=TMG), organic compounds that accumulate to high intracellular dimethylglycine, sarcosine, camitine, N-methyl alanine and levels under osmotic stress and that are compatible with trimethylamino-butyric acid, butyrobetaine, proline betaine the metabolism of the cell’). and other amino acid betaines; 0035. A compatible solute according to the present inven the N-methylated aminoalcohol is preferably selected from tion is thus an organic osmolyte or osmoprotectant. choline and choline derivatives such as choline-o-fosfate, 0036. The expression “glucogenic” as used in this context glycerophosphorylcholine (GPC); refers to compounds that can be converted into glucose through a metabolic pathway that results in the generation of the N-methylated amine oxide is preferably trimethylamine glucose from non-carbohydrate carbon Substrates. oxide (TMAO); 0037 Compatible solutes are organic molecules accumu the N-methylated amidines are preferably selected from ecto lated in high concentrations in the cytoplasm allowing adap ine and hydroxyectoine; and tation of the cell to varying salt concentrations. They are the poly amino hydrocarbon compounds are preferably uncharged, highly water soluble and have to maintain an selected from putrescine, cadaverine, spermine and spermi osmotic equilibrium with the Surrounding medium. Some of dine, as well as polyamine growth factors from eucaryotic these compounds also show secondary functions such as Sup cells. port and protection of macromolecules (e.g. enzymes). The 0.018. The nitrogen containing, non glucogenic, compat cells may accumulate these compounds from the Surrounding ible solute compounds according to the invention are thus medium or by synthesis. Small glucogenic molecules (glyc specifically selected from erol, maltose, trehalose, Sucrose, amino acids, etc.) may also 0019 taurine, function as compatible solutes. Beside these compounds, 0020 choline and choline derivatives, most compatible solutes are N-containing compounds or (0021 trimethylamine-oxide (TMAO), polyols. 0022 ectoine and hydroxyectoine, 0038 N-containing compounds are reported to be gener 0023 the N-methylated aminoacids glycine betaine, ally more potent than others. Cells must be equipped with a dimethylglycine, sarcosine, carnitine, N-methyl ala transporter system to concentrate "cheap' compatible solute nine, trimethylamino-butyric acid, butyrobetaine and compounds from outside. Compatible solutes Such as amino proline betaine, and acids and Saccharides may be used in glucogensis for ATP 0024 the poly amino hydrocarbon compounds production. Higher polyamines are also reported to act as putrescine, cadaverine, anabolic growth regulators in fruits and vegetables and are 0025 spermine and spermidine, involved in shelf life. 0039 Plants are, through their leaves, an enormous area to 0026 and mixtures of any of these. be colonised by microbes. This area is nevertheless non 0027. It is observed that the suitable synergistic com attractive for microbes due to continuous water stress condi pounds in accordance with the invention, for Supplementing tions on the wax layer. Temperature changes and UV irradia the known antimicrobial effect of Silver--Peroxide thus tion are also stress factors for microbes. Pathogenic microbes include “betaine' derivatives. will try to enter the leaves for better conditions. Spores have 0028. It is specifically acknowledged in this context that it to germinate before attacking the plant. Germination of has been proposed in the state of the art spores is a strong energy dependent process and time con 0029 (for instance in CN 17297.87, GB2354771, EP 1 Suming. It is therefore interesting to regularly spray plants 225887, EP 1 036 511, and in cosmetic formulations) with the combination peroxide/silver to kill vegetative to associate silver and peroxides with specific tension active microbes and inhibit sporulation. “betaines', as surfactants. 0040 Compatible solutes, other than urea, do not interact 0030. It has to be stressed that the expression “betaines' in with macromolecules and they have lithe impact on cell func the more broad sense as used in the latter case, is totally tions. This is in contrast to inorganic ions which bind to and different from the expression “betaine', stricto sensu, as used destabilize organic molecules Such as proteins and nucleic in respect of N-methylated aminoacids according to the acids at relatively high concentrations. Compatible solutes US 2011/0212 187 A1 Sep. 1, 2011

are neutral (or Zwitter ionic) at physiological pH. Some bac from the external medium. Accumulation of anti-stress com terial Compatible solutes are anionic but complex with cat patible solutes, as TMG is a better compatible solute than the ions. Cells may use at the same time different compatible polyols, reaches higher intracellular levels and is less energy solutes. They are interchangeable. Trimethylglycine (TMG) demanding than the denovo synthesis of polyols and dipep may replace Sorbitol or other polyols in eukaryotic or tides. Also in fungi, the synthesis of polyols is energy con prokaryotic cells to restore viability under hypertonic condi Suming, especially in Solid-state fermentation where the tions. The use of urea is excluded because it may destabilize water activity is decreasing. Addition of glucogenic compat macromolecules. Unmethylated amino acids are also ible Solute will help these organisms to synthesize new com excluded, because they are reported to all react with silver patible solutes and to Sustain expulsion activity against per ions and are mostly glucogenic or ketogenic. oxides and silver compounds. Most compatible solutes are 0041 Honglucogenic compounds do not realize energetic widespread in kingdoms of the tree of life, some are restricted reserves in the cell. Most of these compounds also show anti-oxidative activity in plants or other biological activities to a small numbers of organisms. TMG, polyamins and ecto different from their osmotic activity. Compatible solutes may ine are able to restore normal turgor pressure in stress condi also show different effect depending on the environmental tions in prokaryotic and eukaryotic cells. conditions. Trehalose protects enzymes at high temperature 0046. Some glucogenic compatible solute may initiate but inhibits them at normal temperature after intracellular microbial growth when the peroxide is exhausted. In most concentration. They may be harmful in the absence of a published in vitro experiments, microbial Suspensions are perturbant (temperature). It is therefore important not to use used and not microbial biofilms which are omnipresent (pro high compatible solute concentrations. They may be espe cess water piping systems, containers, on leaves and roots, on cially harmful if used where their non-osmotic properties are contaminated Surfaces, etc. ...). Biofilm embedded microbes not needed Such as anti-oxidant activity, redox balancing are using "quorum sensing molecules to counteract as one activity, calcium modulation, detoxification activity, counter unit the increasing concentrations of anti-microbial com acting enzymatic inhibition or a compensatory activity. This pounds as antibiotics, disinfectants or pesticides. They make may be different for a microbe and host cell (plant, human or “global decisions to expulse such molecules through the animal). Microbes on leafs, roots or skin are mostly under elaboration of a dense network in the biofilm and the forma osmotic stress while cellular host cells or tissues are under tion of new enzymes. The biofilm becomes stronger and other kinds of stress Such as oxidative stress. stronger and increases in mass. It could be interesting to 0042. The accumulation of osmolytes in crops under mislead the internal microbes using non glucogenic compat drought conditions is often cited in literature. However, field ible solutes combined together with the use of antimicrobial studies examining the association of osmolytes and crop compounds. The Small compatible solute molecules are neu yields have shown no consistent benefit. Besides a speculated tral and penetrate the biofilm matrix. Host tissues under bio benefit of turgor maintenance or root development in order to film matrix (infected tissues) could also benefit from the use reach water, no clear increase in crop yield was observed. The of such compatible solutes and better resist oxidative stress secondary activity of the osmolyte (anti-oxidant) results in a conditions. The normalization of turgor on the microbial kind of water deficit tolerance in plants. Hyper accumulation cytoplasmic membrane could increase the access of peroX of compatible solutes may even be a symptom of injury. ides and silver compounds. We discovered that use of non 0043. Similar effects are seen in microbes using non glucogenic compatible solutes is beneficial for the anti-mi glucogenic compatible solutes: maintenance of turgor pres crobial activity of the peroxide-silver combination in biofilm Sure but no clear stimulation of growth under osmotic stress treatment. conditions. Cells normally synthesize compatible solutes as 0047. In a medium containing non glucogenic compatible reaction on an osmotic up chock but degrade the osmolytes Solutes, peroxides are able to destroy the organic matrix into following and osmotic down chock. The initial response is nonharmful Substances through oxidation. Silver compounds mostly quicker when extracellular compatible solutes are activate their anti-microbial activity which is desirable due to taken up or released to the medium via transport systems the quick consumption of the peroxides in the presence of instead of de novo synthesis. organic material. We also detected that the use of chelators 0044) When we look carefully at the literature concerning Such as EDTA, polyacrylates and phosphonates, decreased the effect of compatible salutes on microbes or microbial the activity of the peroxide-silver combination. This is oppo growth we encounter a lot of contradictions. This is mostly site to the use of chelators in various publications and patents. due to the use of complex growth media in which already a lot Nonglucogenic compatible solutes also show interesting sec of compatible solutes are incorporated. Simple molecules ondary activities, different from ATP generation, besides their Such as mono- and disaccharides, amino acids and polyols are osmolytic activity. This activity is beneficial to the host tissue. present in most culture media and have not only a function as compatible solute but in the first place an energy generating 0048. Without intending to link the invention to any spe function. They are glucogenic and generate ATP. They con cific explanation, it is Supposed that the use of compatible tribute not only to osmoregulation but also to ATP-dependent Solutes may be appropriate for different reasons: expulsion of peroxides and silver compounds and to the 0049 A) Activation of the antimicrobial activity, prob absorption or expulsion of non-glucogenic compatible sol ably resulting from the normalization of the cytoplas utes. matic turgor 0045 Strains of Staphilococcus aureus show a significant 0050 B) Decrease of the oxidative stress in biological increase in membrane fluidity during the growth phase but host tissues decreasing fluidity entering in the stationary phase and 0051 C) Compatible solutes are non toxic and disap decreasing sensitivity to biocidals. A lot of microbes are able pear in the medium because most organisms in the envi to accumulate N-methylated or polyamine compatible solutes ronment are able to concentrate these molecules. US 2011/0212 187 A1 Sep. 1, 2011

Selected compatible solutes are taken up by many organ 0058. The composition is ecologically friendly and phar isms and are able to replace internally produced solutes maceutical preparations can also be used for topical applica under stress conditions tions in humans and animals. 0.052 D) Residual compatible salutes are taken up by 0059. According to a further preferred feature of the symbiotic organisms. invention, the silver source of the aantimicrobial composition is most appropriately selected from 0053. It has been found, in particular, that the use of non a) metallic silver, colloidal silver or silver nanoparticles glucogenic compatible solutes does not stimulate microbial b) nano particles or silver chloride nano par growth at low concentrations but on the contrary activate ticles made in situ on a natural carrier containing chloride ions anti-microbial activity of the peroxide-silver combination. or through addition of chloride ions, colloidal silver salt/ Glucogenic salutes are mostly represented by mono- and carrier, silver oxide/carrier or ionic silver/carrier (carriers are disaccharides and amino acids. low in thiols) or mixtures thereof, 0054 Plants under biotic and abiotic stress are more vul c) silver nitrate, silver fluoride, silver chloride, silver oxide, nerable to infection and show a decrease in growth and devel silver Zeolite, silver sodium hydrogen Zirconium phosphate, opment and therefore need to elaborate higher compatible silver carbonate, silver sulphide, silver selenate, silver sul Solute concentrations for the entire plant including the roots phate or mixtures thereof. and Surrounding symbiotic organisms. Trehalose, a non d) silver amino acids complexes reducing disaccharide found in different organisms, is syn e) , , silver chromate, silver thesized under Such conditions as compatible solute, stabi hydroxide, silver iodate, silver molybdate, silver oxalate, sil lizes enzymes and membranes and protects structures from verperchlorate, silver sulfardiazine, mild silver protein, silver desiccation. It is present in higher concentrations in plants thiosulfate or mixtures thereof. with diseases or in colonized plants. f) silver salts of carboxylic acids and dicarboxylic acids or 0055 Compatible solutes are synthesized under stress mixtures thereof, or mixtures of any of these conditions but are again destroyed under normal conditions. 0060. Further preferred features of the antimicrobial com Osmolyte concentrations in plants can also be used as an position of the invention involve that indicator of finished germination (malt quality, preharvest 0061 the concentration (w/v) of the nitrogen contain sprouting in seeds, etc....). Oxidative stress due to present of ing compound or the compatible solute is lower than the iron and copper ions in water Supply stimulates the produc peroxide concentration and not more than 100 times tion of polyamines in plants. higher than the silver concentration; 0056. It was also found that compatible solutes interfere 0062 the total peroxide concentration ranges from 60% with the action of silver/peroxide on the pathogens. The sil to 0.1% peroxide (in the concentrated solution); Ver/peroxide also shows an anti-oxidative activity on the 0.063 the organic peroxide is selected from a hydroper plants. This new combination is also useful in animal farming oxide, a peroxyacid, a perester type, a peroxy-methoxy (poultry, pigs, horses, etc.) resulting in a higher production, a acid or a peroxy-phenylacid, or combinations thereof, better feed-conversion besides the lowering effect on micro whereas bial pressure resulting in animal diseases. The nonglucogenic 0.064 the peroxyacid is preferably a peroxycarboxylic compatible solutes are used simultaneous on the pathogens acid, a peroxydicarboxylic acid, a peroxytricarboxylic and the host. The compatible solutes are used at limited con acid, a peroxy hydroxy acid or esters thereof and com centrations not inducing microbial growth resulting in the binations thereof exhausting of the peroxides due to the microbial formation of 0065 the peroxyacid is peroxy lactic acid, peroxypro catalase and peroxidase. The presence of a silver compound pionic acid, peroxy citric acid, peroxyacetic acid, per enhances the activity of the peroxide and reacts with peroxide formic acid, perbenzoic acid, ethaneperoxoic acid, per inactivating enzymes. The combinations of peroxide, silver oxy methoxyacetic acid, peroxyphenyl acetic acid, compounds and compatible solutes, used at appropriate con mono or diperoxydicarboxylic acid, mono or di-esters centrations, are less harmful than most pesticides or biocides. thereof and combinations thereof. The amount of residues on fruits and vegetables are neglect 0.066 the peroxycarboxylic acid is obtained through able. Especially children are susceptible to the harmful oxidation, with hydrogen peroxide or a hydroperoxide, effects of pesticides, cleaning products and tension active of lactic acid, propionic acid, citric acid, oxalic acid, compounds. Higher occurrence of cancer, bird defects, leu acetic acid, formic acid, benzoic acid, malonic acid, kaemia were found in children with early exposure to pesti glutaric acid, pimelic acid, Suberic acid. Succinic acid, cides. In general, pesticides, biocides, and tensioactive com aZeleic acid, adipic acid, Sebasic acid, or derivatives pounds provoke oxidative stress in their host. It is therefore thereof and esters thereof; interesting to add bioactive compatible solutes (non gluco 0067 the inorganic peroxide is an inorganic peroxyacid genic at low concentrations) to the microbes, probably nor or a salt thereof, whereas malizing turgor pressure and thereby increasing the target 0068 the peroxyacid is: Surface for the biocides. At the same time, a positive action is 0069 a) peroxymonosulfuric acid or peroxy disulfuric noticed on the host tissues, lowering the stress or conse acid, Sodium, potassium or ammonium salts thereof and quences of inflammation or tissue damage. mixtures thereof. 0057 The new antimicrobial compositions according to 0070 b) peroxynitrous acid and salts thereof the invention can be diluted in all kinds of water and such a 0071 c) permitrate and salts thereof dilution is useful as disinfectant, antimicrobial biocide, 0.072 d) urea peroxide or urea hydrogen peroxide hygienic compound, pesticide (algaecide, bactericide, fungi 0.073 e) percarbonate (example: sodiumpercarbonate) cide, nematocide and virucide), decontamination compound 0074 f) calcium, Sodium, barium or magnesium peroX or as anti-biofouling compound. ides and mixtures thereof US 2011/0212 187 A1 Sep. 1, 2011

0075 g) hydrogen peroxide or activated hydrogen per Example 6 oxide (hydrogen peroxide complexes of inorganic salts) 0076 h) permanganate 0099 H2O2: 50% 0077 i) perboric acid and salts thereof (example: 01.00 AgNO3:300 mg Ag/litre Sodium perborate), or mixtures of any of these 0101 Carnitine: 0.25% (w/v) 0078 the concentration of the inorganic peroxide varies Example 7 between 50% and 0.1% (w/v) peroxide (in the concen trated composition); 01.02 H2O2: 40% 0079 the silver concentration ranges from 10 mg/liter (0103) Silver(II) oxide (AgO) as colloid: 250 mg Ag/litre and 1000 mg/liter (in the concentrated composition); 0104 Potassium monopersulfate: 5% 0080. The invention also specifically relates to the use of 01.05 TMG: 0.5% (w/v) an antimicrobial composition as defined here above, as a disinfectant, antimicrobial biocide, hygienic compound, Example 8 algaecide, bactericide, antimicrobial pesticide, fungicide, 01.06 H2O2: 40% acidified with 0.1N HC1 nematocide, Virucide, decontaminating or anti-biofouling 01.07 AgCl (450 mg Ag/litre) made in situ on a natural compound in a ready to use formulation or a concentrated carrier formulation to be diluted in water or an aqueous solution, 0108 TMG: 2.5% (w/v) emulsion or Suspension. 0081. The invention also relates to the use of such antimi Example 9 crobial composition as a topical composition concentrated or after dilution or as ingredient in the preparation of ointments, 0109 H2O2: 40% gels, cremes, or any acceptable topical pharmaceutical prepa 0110 Silverthiosulfate (350 mg Ag/litre) ration. 0111 TMAO: 0.1% (w/v) 0082. The invention also relates to such antimicrobial Example 10 compositions comprising one or more additional fungal spore inhibiting compounds such as polyols, terpenes, carboxylic O112 H2O2: 40% acid, Sugar acids, boric acids, metal salts and combination 0113 Peracetic acid: 1.5% thereof. 0114 Silver nitrate: 300 mg Ag/litre 0083. Further features and details of the invention will 0115 TMG: O.2% appear from the following specific, non limited examples of antimicrobial compositions and antimicrobial applications Example 11 according to the invention. 0116 H2O2: 40% 0117 Peracetic acid: 1.0% EXAMPLES OF ANTIMICROBIAL 0118 Silver nitrate: 360 mg Ag/litre COMPOSITIONS 0119) PEG4OO or PEG8OO: 10% Example 1 0120 TMAO: O.2% 0084 H2O2: 50% Example 12 0085 AgNO3: 350 mg Ag/litre 0121 H2O2: 50% 0086 TMG: 0.05% (w/v) 0122) Silver nitrate: 200 mg Ag/litre 0123 TMG: 0.05% Example 2 0.124. This solution can be used (as spray) after dilution in 0087 H202:38-40% purified water for decontamination and disinfection (nano or 0088 Colloidal Silver/carrier containing 350 mg Ag/litre micro droplets) (low thiol carrier) Example 13 0089 TMG: 0.1% (w/v) 0.125 H2O2: 40% Example 3 0126 Potassium monopersulphate: 10% O127 Silver nitrate: 250 mg Ag/litre 0090 H2O2: 50% 0128 PEG200: 10% 0091 AgNO3: 250 mg Ag/litre 0129 TMG: O.1% 0092 TMG: 0.2% (w/v) Example 14 Example 4 0.130 H202: 50% 0093 H2O2: 40% I0131 Silver chloride made in situ on a gum carrier: 380 0094) AgNO3: 350 mg Ag/litre mg Ag/litre 0095 TMAO: 0.1% (w/v) 0132 TMG: 1% Example 5 Example 15 0.133 H202: 50% 0.096 H2O2: 40% 0.134 Silver chloride made in situ on a thiol low natural 0097 AgNO3: 400 mg Ag/litre carrier: 340 mg Ag/litre 0098 Taurine: 0.3% (w/v) 0.135 GPC: 0.05% US 2011/0212 187 A1 Sep. 1, 2011

Example 16 Example 22 0.136 H202: 50% (0168 45% H.0, 0.137 Colloidal Silver/carrier containing 325 mg Ag/litre (0169. 2.5% polyol (carrier must be thiollow) (0170 2% boric acid 0138 TMG: 1% 0171 560 mg Ag/litre as AgNO (0172 0.3% menthol or eugenol Example 17 0173 0.3% TMG 0139 H202: 7.9% Example 23 0140 Colloidal Silver/carrier containing 70 mg Ag/litre (carrier must be thiollow) (0174 45% H.0, 0141 TMG: 0.1% (0175 3% boric acid 0142. All antimicrobial compositions of the above 0176 1% galacturonic acid examples, containing the mentioned peroxides, have been (0177 550 mg Ag/litre as AgNO, used as solution diluted 200 times (70 times in the case of 0178. 1% sorbose Example 17) in source water (containing 150 mg/l ), 0179 0.3% TMG and have resulted in at least a 5 log titer reduction of Staphy lococcus aureus at 25 degrees Celsius after 2 hours incuba Examples of Antimicrobial Applications tion. Example 24 Example 18 0180. The antimicrobial composition of example 2 (com 0143 Calcium peroxide: 35% position labelled “SN025-B”) was applied on rice for the 0144. Silveroxide: 0.2% Silver control of Rice Blast, Sheath Blight, Brown Spot (Pyricularia (0145 TMG: 3% (w/v) Oryzae, Rhizoctonia sp., Helminthosporium sp.) 0146 Phosphate buffer: 20% 0181 Measurements were made for an average of two 0147 Sodium percarbonate: 10% trials. 0148 Silica until 100% 0182 3 applications at 14 days interval 0149 Prepare a 2% solution of this mixture before use for 0183 Assessments in December 200745-50 days sowing. soil treatment or on crops. First dilute in demineralised water. 0.184 The results are illustrated in diagram 1: 0185. Strong infection in the untreated (64%, 52% & 50% Example 19 for Rice Blast, Sheath Blight & Brown Spot respectively). 0186. The composition SN025-B achieved 33-55% effi 0150. Propylene glycol: 55% 0151. Sodium perborate monohydrate: 25% oit (44-60% for a reference program with 2 triazoles) 0152 Silver nitrate: 0.3% Silver 0153 TMG:3% Example 26 0154 Add fumed silica until 100% 0188 The same composition labelled “SN025-B” was (O155 Dilute in water just before use. applied for the control of Leaf Spot (Cyclonium oleaginum) 0156 Inorganic peroxides may be used in aquaculture to on Olive trees prevent growth of anaerobes and other bacteria; in poultry for (0189 Assessments on 8 May 2008 (% of defoliation on decontamination of fodder, productivity increase and the 200 leaves) improvement of egg quality; in cattle: inhibition of diarrhoea 0190. Applications in preventive spray schedule (2 appli in calves and as anti-microbial compound, normalizing ali cation in Autumn and 2 applications in Spring) mentary track and digestion. 0191 The results are illustrated in diagram 2: 0157 For dental use: teeth bleaching and as antimicrobial (0192 SN025-B achieved 47% efficacy (comparable to compound Copper) Example 20 Example 26 0158 45% H0, (0193 The same composition labelled “SN025-B” was 0159) 5% polyol applied for the control of Septoria Leaf Spot (Septoria sp.) on (0160 550 mg Ag/litre as AgNO, Winter Wheat (0161 0.2% TMG 0194 Assessments on 2 May 2008 (% of Septoria on L4) (0195 One application at TO (7 Apr. 2008) Example 21 0196. The results are illustrated in diagram 3: (0162 45% H0. 0.197 SN025-B achieved 58% efficacy (0163 2.5% polyol Example 27 (0164 0.3% malic acid (0165 0.15% peracetic acid (0198 The same composition labelled “SN025-B” was 0166 580 mg Ag/litre as AgNO applied for the control of Black Spot (Vinaequalis) on Apple 0167 0.3% TMG (0199 Assessments on 14 Dec. 2007 and 3 Feb. 2008 US 2011/0212 187 A1 Sep. 1, 2011

0200. Applications in preventive spray schedule (at 7-14 4. Antimicrobial composition according to claim 1, days interval during primary contaminations, then at 14-28 wherein the total peroxide concentration ranges from 60% to days) 0.1% peroxide. 0201 The results are illustrated in diagram 4: 5. Antimicrobial composition according to claim 1, (0202 SN025-B achieved 65% efficacy (comparable to wherein the organic peroxide is a hydroperoxide, a peroxy organic fungicides) acid, a perester type, a peroxy-methoxy acid or a peroxyphe nyl acid, or combinations thereof. Example 28 6. Antimicrobial composition according to claim 5. wherein the peroxyacid is a peroxycarboxylic acid, prefer 0203 The same composition labelled “SN025-B” was ably peroxy lactic acid, peroxypropionic acid, peroxy citric applied for the control of Botrytis cinerea on Strawberry acid, peroxyacetic acid, performic acid, perbenzoic acid, 0204 Assessments at 7 days interval, starting 7 days after ethaneperoxoic acid, peroxy methoxyacetic acid, or peroX last application (4 applications in total) yphenyl acetic acid; a mono or diperoxydicarboxylic acid; a 0205 The results are illustrated in diagram 5: peroxytricarboxylic acid; a peroxy hydroxy acid; or esters 0206 SN025-B achieved 80% efficacy (comparable to thereof and combinations thereof, whereas the peroxycar organic fungicides) boxylic acid is preferably obtained through oxidation, with 0207. Yield was 18.8% higher versus untreated and 7.9% hydrogen peroxide or a hydroperoxide, of lactic, propionic higher versus thiophanate-methyl treated plots acid, citric acid, oxalic acid, acetic acid, formic acid, benzoic 1. Antimicrobial composition comprising a combination of acid, malonic acid, glutaric acid, pimelic acid, Suberic acid, at least one organic and/or inorganic peroxide, Succinic acid, azeleic acid, adipic acid, Sebasic acid, or at least one silver source, and derivatives thereof and esters thereof. at least one nitrogen containing compound which is a non 7. Antimicrobial composition according to claim 1, glucogenic compatible solute and which is selected from wherein the inorganic peroxide is a weak or strong inor taurine, ganic peroxyacid or a salt thereof. choline and choline derivatives, 8. Antimicrobial composition according to claim 7. trimethylamine-oxide (TMAO), wherein the peroxyacid is: ectoine and hydroxyectoine, a) peroxymonosulfuric acid or peroxy disulfuric acid, the N-meitlylated aminoacids glycine betaine, dimethylg Sodium, potassium or ammonium salts thereof and mix lycine, sarcosine, carnitine, N-methylalanine, trimethy tures thereof lamino-butyric acid, butyrobetaine and proline betaine, b) peroxynitrous acid and salts thereof and c) permitrate and salts thereof the poly amino hydrocarbon compounds putrescine, d) urea peroxide or urea hydrogen peroxide cadaverine, spermine and spermidine. e) percarbonate 2. Antimicrobial composition according to claim 1, com f) hydrogen peroxide or activated hydrogen peroxide prising a silver source selected from g) permanganate a) metallic silver, colloidal silver or silver nanoparticles h) perboric acid and salts thereof, b) silver chloride nano particles or silver chloride nano or mixtures of any of these particles made in situ on a natural carrier containing 9. Antimicrobial composition according to claim 1, chloride ions or through addition of chloride ions, col wherein the silver concentration ranges from 10 mg/liter and loidal sliver salt? carrier, silver oxide/carrier or ionic sil 1000 mg/liter. ver? carrier or mixtures thereof, 10. Antimicrobial composition according to claim 1, used c) silver nitrate, silverfluoride, silver chloride, silver oxide, as a disinfectant, antimicrobial biocide, hygienic compound, silver Zeolite, silver Sodium hydrogen Zirconium phos algaecide, bactericide, antimicrobial pesticide, fungicide, phate, silver carbonate, silver Sulphide, silver selenate, nematocide, Virucide, decontaminating or anti biofouling silver sulphate or mixtures thereof. compound in a ready to use formulation or a concentrated d) silver amino acids complexes formulation to be diluted in water or an aqueous solution, e) silver iodide, silver chlorate, silver chromate, silver emulsion or Suspension. hydroxide, silver iodate, silver molybdate, silver 11. Antimicrobial composition according to claim 1, used oxalate, silver perchlorate, as a topical composition concentrated or after dilution or as silver sulfardiazine, mild silver protein, silver thiosulfate ingredient in the preparation of ointments, gels, cremes, or or mixtures thereof any acceptable topical pharmaceutical preparation. f) silver salts of carboxylic acids and dicarboxylic acids or 12. Antimicrobial composition according to claim 1, com mixtures thereof, or mixtures of any of these prising one or more additional fungal spore inhibiting com 3. Antimicrobial composition according to claim 1, pounds such as polyols, terpenes, carboxylic acid, Sugar wherein the concentration (w/v) of the nitrogen containing acids, boric acids, metal salts and combination thereof compound is lower than the peroxide concentration and not more 100 times higher than the silver concentration. c c c c c