TWI526157B - Antimicrobial formulation - Google Patents

Description

Antimicrobial formulation

A synergistic antimicrobial formulation comprising a mixture of an organic acid, an alpha, beta-unsaturated fatty aldehyde, and an antimicrobial steroid.

A recent report by Georgetown University's Pew Charitable Trusts points out that food-borne illness costs US$152 billion annually in health-related costs. According to the Centers of Diseases Control and Prevention, the number of people suffering from foodborne illnesses in the United States is 76 million and 5,000 deaths per year (Thomson Reuters 2010, 3/10/2010).

Therefore, it is desirable to find more natural and/or organic antimicrobial agents, which has led to a great deal of research; and because of the low commercial availability of these new natural/organic products, the cost of the raw materials has increased.

Several new antimicrobial agents have been found in plants. We have found that when grass or other plants are damaged by harvesting, trimming or exposure to pathogens, the lipoxygenase pathway will be initiated, resulting in the production of compounds exhibiting antimicrobial activity. Lipoxygenase is a widely distributed enzyme that catalyzes the oxidation of unsaturated fatty acids to form fatty acid hydroperoxides. As a matrix of synthetic compounds, this fatty acid hydroperoxide plays an important role in plant defense. (Kishimoto, K.; Matsui, K.; Ozawa, R.; Talcabayashi, J. "Direct fungicidal activities of C6-aldehydes are important constituents for defense responses in Arabidopsis against Botrytis cinerea ." Phytochemistry 2008., v.69., 2127-2132; Gardini, F.; Lanciotti, R.; Guerzoni, ME "Effect of trans -2-hexenal on the growth of Aspergillus flavus in relation to its concentration, temperature and water activity." Letters in App. Microbiology 2001, v. 33, 50-55).

The aldehydes are represented by the formula RCHO wherein R can be H or an aliphatic, aromatic or heterocyclic group. The α,β-unsaturated fatty aldehydes of 3 to 10 carbon units have important commercial significance. They are moderately soluble in water and their solubility decreases with increasing molecular weight.

α,β-unsaturated fatty aldehydes include acrolein, trans-2-butenal, 2-methyl-butenal, 2-methyl-(E)-2-butenal, 2-pentene Aldehyde, trans-2-hexenal, trans-2-hexen-1-ol, 2-methyl-2-pentenal, 2-isopropylpropenal, 2-ethyl-2-butene Alkenal, 2-ethyl-2-hexenal, (Z)-3-hexenal, 3,7-dimethyl-6-octenal, 3,7-dimethyl-2,6- Octadialdehyde, (2E)-3,7-dimethyl-2,6-octadienal, (2Z)-3,7-dimethyl-2,6-octadienal, trans- 2-nonenal, (2E, 6Z)-nonadienal, 10-undecaldehyde, 2-dodecenal and others. The present invention includes C3 to C12 alpha, beta-unsaturated fatty aldehydes.

Fats and phospholipids cleave to form aldehydes of three, six and nine carbon atoms, including (2E,6Z)-nonadienal, trans-2-nonenal and trans-2-hexenal . These compounds are produced by a combination of two different enzymes of the plant lipoxygenase (LOX) pathway. In the first reaction, LOX catalyzes the oxidation of polyunsaturated fatty acids, such as linoleic acid and linoleic acid, which in turn form 9- or 13-hydroperoxides. These compounds are very unstable and are then cleaved into aldehydes and oxoacids by enzyme hydroperoxide lyase. From the 9-hydroperoxy group, trans-2-nonenal and (2E,6Z)-nonadienal are obtained, and from the 13-hydroxide derivative, hexenal and trans-2-hexene are obtained. aldehyde. In the second reaction, the aldehydes can be converted to the corresponding alcohols by the action of alcohol dehydrogenases (Hubert, J.; Munzbergova, Z.; Santino, A. "Plant volatile aldehydes as natural insecticides against stored-product beetles. Pest Manag. Sci . 2008, v. 64, 57-64).

The volatile compound used in the present invention is trans-2-hexenal which is a 6 carbon, double bond aldehyde, C ₆ H ₁₀ O, and has a molecular weight of 98.14. According to the European Council Directive 88/388/EEC and the US FDA 21 CFR 101.22, natural or synthetic trans-2-hexenal, also known as folate, is considered a natural seasoning. .

Trans-2-hexenal is present in many edible plants such as apples, pears, grapes, strawberries, kiwis, tomatoes, olives and the like. Existing studies have successfully used these plants and their extracts to find new antimicrobial agents. For example, cashew pears are effective against Helicobacter pylori and S. cholerasuis (50-100 ug/ml). The two main components were found to be cashew acid and trans-2-hexenal. The minimum inhibitory and minimal bactericidal activity concentrations of trans-2-hexenal were determined to be 400 and 800 ug/ml, respectively (Kubo, J.; Lee, JR; Kubo, I. " Anti-Helicobacter pylori Agents from the Cashew Apple.” J. Agric. Food Chem . 1999, v. 47, 533-537; Kubo, I. And K. Fujita, “Naturally Occurring Axiti-Salmonella Agents” J. Agric. Food Chem . 2001, v. 49, 5750- 5754). Kim and Shin found that trans-2-hexenal (247 mg/L) is effective against B. cereus , S. typhimurium , V. parahaemolyticus , single L. monocytogenes , S. aureus , and E. coli 0157:H7 (Kim, YS; Shin, DH "Volatile Constituents from the Leaves of Callicarpa japonica Thunb And their Antibacterial Activities." J. Agric. Food Chem. 2004, v. 52, 781-787). Nakamura and Hatanaka demonstrated that (3E)-hexenal can effectively control Staphylococcus aureus , E. coli and Salmonella typhimurium ("Green-leaf-derived C6" at 3-30 ug/ml -aroma compounds with potent antibacterial action that act on both gram-negative and gram-positive bacteria." J. Agric. Food Chem. 2002, v. 50 no, 26, 7639-7644). Trans-2-hexenal completely inhibits the proliferation of Pseudomonas ( P. syringae ) lesions (570 ug/L air) and E. coli (930 ug/L air) (Deng, W.; Harnilton- Kemp, T.; Nielsen, M.; Anderson, R.; Collins, G.; Hilderbrand, D. "Effects of Six-Carbon Aldehydes and Alcohols on Bacterial Proliferation." J, Agric. Food Chem. 1993, v. 41 , 506-510). Trans-2-hexenal was observed to effectively inhibit the growth of Phoma mycelium at 250 ug/ml (Saniewska, S. and M. Saniewski, 2007. "The effect of trans- 2-hexenal and trans-2-nonaenal on the mycelium growth of Phoma narcissi in vitro, Rocz. AR, Pozn. CCCLXXXIII, Ogrodn. V. 41, 189-193"). In a study to control mold in fruits, trans-2-hexenal was not phytotoxic to apricot at a concentration of 40 uL/L, but phytotoxic to peach and nectarine ((Neri, F., M) Mari, S. Brigati and P. Bertolini, 2007, "Fungicidal activity of plant volatile compounds for controlling Monolinia laxa in stone fruit" Plant Disease v. 91, no. 1, 30-35). Trans-2-hexene Aldehyde (12.5 uL/L) is effective in controlling penicillium-producing Penicillium expansum (Neri, F.; Mari, M.; Menniti, A,; Brigati, S.; Bertolini, P. "Control of Penicillium" Expansum in pears and apples by trans-2-hexenal vapours.” Postharvest Biol and Tech . 2006, v. 41, 101-108. Neri, F.; Mari, M.; Menniti, AM; Brigati, S. “Activity of Trans-2-hexenal against Penicillium expansum in ' Conference'pears." J. Appl. Micrbiol 2006, v. 100, 1186-1193). Hamilton-Kemp et al. proposed trans-2-hexenal vapor to inhibit gray mold Germination of disease spores and apple pollen ( J. Agric. Food Chem . 1991, v. 39, no. 5, 952-956).

U.S. Patent Application Serial No. 2007/0087094 teaches the use of at least two GRAS compounds having microbicidal activity in combination with less than 50% alcohol (isopropanol or isopropanol/ethanol) as a microbicide. Trans-2-hexenal may be one of the GRAS compounds. Archbold et al. also observed that the use of trans-2-hexenal for the use of 0.86 or 1.71 mmol (100 or 200 microliters of pure compound per 1.1 L of the container) after steaming for two weeks indicates that it can be controlled. Mold (Archbold, D.; Hamilton-Kemp, T.; Clements, A,; Collins, R. "Fumigating' Crimson Seedless' Table Grapes with (E)-2-Hexenal Reduces Mold during Long-term Postharvest Storage." HortScience 1999, v. 34, no. (4, 705-707)).

U.S. Patent 5,698,599 teaches a method for inhibiting the production of mycotoxins in foodstuffs by treatment with trans-2-hexenal. Trans-2-hexenal completely inhibits A. flavus , P. notatum , A. alternate , and Fusarium oxysporum at a concentration of 8 ng/L air. Growth of ( F. oxysporum ), Cladosporium species , B. subtitus and A. tumerfaciens . Comparing trans-2-hexenal with citral in controlling yeast (10 ⁵ CFU/bottle) in beverages, it was found that 25 ppm trans-2-hexenal plus heat treatment (56 ° C, 20 minutes) is equivalent to 100 - 120 ppm of citral. In beverages, 35 ppm trans-2-hexenal is required to stabilize if not heat treated (Belletti, N.; Kamdem, S.; Patrignani, F.; Lanciotti, R.; Covelli, A.; Gardini, F. "Antimicrobial Activity of Aroma Compounds against Saccharomyces cerevisiae and Improvement of Microbiological Stability of Soft Drinks as Assessed by Logistic Regression." AEM . 2007, v. 73, no. 17, 5580-5586). Trans-2-hexenal is not only used as a microbicide, but is also observed to be effective in controlling insects. Volatile substances (ie trans-2-hexenal) are effective against beetles such as Tibolium castaneum, Rhyzopertha dominica, Sitophilus granaries, Sitophilus orazyzae and Cryptolestes perrugineus (Hubert, J.; Munzbergova, Z.; Santino, A." Plant volatile aldehydes as natural insecticides against stored-product beetles." Pest Manag. Sci . 2008, v, 64, 57-64). U.S. Patent 6,201,026 teaches an organic aldehyde having three or more carbons which can control aphids.

Several patents teach the use of trans-2-hexenal as a fragrance or fragrance. U.S. Patent 6,596,681 teaches the use of trans-2-hexenal as a fragrance on a wipe for surface cleaning. The use of essential oils or terpenoids (e.g., trans-2-hexenal) as perfumes for antimicrobial products is taught in U.S. Patent No. 6,387,866, U.S. Patent No. 6,960,350, and U.S. Patent No. 7,638,114. U.S. Patent No. 6,479,044 discloses an antibacterial solution comprising an anionic surfactant, a polycationic antimicrobial agent and water, wherein the essential oil is added as a perfume. The perfume may be a terpenoid such as trans-2-hexenal or other type of terpenoid. A microemulsion for antimicrobial purposes is disclosed in U.S. Patent No. 6,323,171, U.S. Patent No. 6,121,224, and U.S. Patent No. 5,911,915, which contain a cationic surfactant in which essential oil is added as a perfume. The fragrance may be various anthraquinones including trans-2-hexenal. U.S. Patent 6,960,350 discloses an antifungal fragrance wherein synergistic effects (e.g., trans-2-hexenal with benzaldehyde) are found when a combination of different terpenes is used.

The mode of action of trans-2-hexenal is considered to be: reacting trans-2-hexenal with a thiol moiety or a cysteine residue, or forming a Schiff base with the amino group of the peptide and protein, thereby altering the cell membrane ((Deng, W.; Hamilton-Kemp, T.; Nielsen, M.; Anderson, R.; Collins, G.; Hilderbrand, D. "Effects of Six-Carbon Aldehydes and Alcohols on Bacterial Proliferation." J. Agric Food Chem . 1993, v, 41, 506-510). Trans-2-hexenal is reported as a surfactant that may penetrate across the plasma membrane through passive diffusion. Once inside the cell, its alpha, beta-no The saturated aldehyde moiety reacts with a biologically important nucleophilic group. The alpha, beta-unsaturated aldehyde moiety is recognized to react with the thiol group, primarily by 1,4-addition under physiological conditions (Patrignani, F,;Lucci,L,;Belletti,N.;Gardini,F.;Guerzoni,ME;Lanciotti,R. "Effects of sub-lethal concentrations of hexanal and 2-( E )-hexenal on membrane fatty acid composition and volatile Compounds of Listeria monocytogenes, Staphylococcus aureus, Salmonella enteritidis and Escherichia coli.” Internatio Nal J. Food Micro. 2008, v. 123, 1-8).

Trans-2-hexenal is a phospholipase D inhibitor, an enzyme that catalyzes the hydrolysis of membrane phospholipids during the ripening or ripening of many types of fruits and vegetables. This suggests that trans-2-hexenal may inhibit ripening (U.S. Application Serial No. 2005/0031744 A1). This patent teaches that trans-2-hexenal can inhibit Salmonella and Staphylococcus aureus due to the hydrophobicity and hydrogen bonding present in the partition of the lipid bilayer. The destruction of the electron transport system and the perturbation of cell membrane permeability have been proposed as other modes of action (Gardini, F.; Lanciotti, R.; Guerzoni, ME "Effect of trans-2-hexenal on the growth of Aspergillus flavus in relation to its Concentration, temperature and water activity.” Letters in App. Microbiology . 2001, v. 33, 50-55). The inhibition of extended Penicillium may be due to the destruction of the fungal cell membrane of conidia in germination (Neri, F.; Mari, M.; Menniti, A.; Brigati, S.; Bertolini, P. “Control of Penicillium expansum in pears and Apples by trans-2-hexenal vapours.” Postharvest Biol and Tech . 2006, v. 41, 101-108; Neri, F.; Mari, M.; Menniti, AM; Brigati, S. “Activity of trans-2- Hexenal against Penicillium expansum in 'Conferences pears." J. Appl. MicrbioL 2006, v . 100 , 1186-1193).

Studies have compared trans-2-hexenal and other similar compounds. Deng et al. showed that the unsaturated volatiles trans-2-hexenal and trans-2-hexen-1-ol showed better inhibition than the saturated volatiles hexanal and 1-hexanol (Deng , W.; Hamilton-Kemp, T.; Nielsen, M.; Anderson, R.; Collins, G.; Hilderbrand, D. "Effects of Six-Carbon Aldehydes and Alcohols on Bacterial Proliferation." J. Agric, Food Chem 1993, v. 41, 506-510). Trans-2-hexenal is more active than hexanal, furfural and trans-2-octenal against all ATCC bacterial strains (Bisignano, G.; Lagana, MG; Trombetta. D.; Arena. S.; Nostro, A.; Uccella, N.; Mazzanti, G.; Saija, A. "In vitro antibacterial activity of some aliphatic aldehydes from Olea europaea L." FEMS Microbiology Letters. 2001, v. 198, 9-13 ). Others have found (E)-2-hexenal over hexanal, 1-hexanol, (E)-2-hexen-1-ol and (Z)-3-hexene by measuring several types of mold. -1 - Alcohol has a lower minimum inhibitory concentration of fungal production, essentially aldehydes > ketones > alcohols (Andersen, RA; Hamilton-Kemp, T.; Hilderbrand, DF; McCraken Jr., CT; Collins, RW Fleming, PD Structure-Antifungal Activity Relationships among Volatile C ₆ and C ₉ Aliphatic Aldehydes, Ketones, and Alcohols. J. Agric. Food Chem, 1994, v. 42, 1563-1568). Trans-2-hexenal and hexanoic acid are more effective than hexanol in inhibiting Salmonella (Kubo, I. And K. Fujita, "Naturally Occurring Anti- Salmonella Agents." J. Agric. Food Chem. 2001, v. 49, 5750-5754).

Muroi et al. proposed that trans-2-hexenal exhibits broad-spectrum antimicrobial activity but itself (50-400 μg/mL) is generally not considered to have sufficiently strong biological activity in practical applications (Muroi, H.; Kubo, A.; Kubo, I. "Antimicrobial Activity of Cashew Apple Flavor Compounds," J. Agric. Food Chem. 1993, v. 41, 1106-1109). Studies have shown that trans-2-hexenal can enhance the efficacy of certain types of antimicrobial agents. Several patents teach the use of aminoglycoside avidin enhancers (US 5,663,152) and enhancers of polymyxin antibiotics (US 5,776,919 and US 5,587,358). Such enhancers may include guanidine, anethole, 3-methylindole, 2-hydroxy-6-R-benzoic acid or trans-2-hexenal. When trans 2-heptenal (eptenal), trans-2-nonenal, trans-2-nonenal and (E,E)-2,4-decadienal (according to 1:1: A 1:1 ratio) A strong synergistic effect was observed in anti-ATCC and clinically isolated microbial strains (Bisignano, G.; Lagana, MG,; Trombetta, D.; Arena, S) .; Nostro, A.; Uccella, N.; Mazzanti, G.; Saija, A. "In vitro antibacterial activity of some aliphatic aldehydes from Olea europaea L." FEMS Microbiology Letters. 2001, v. 198, 9-13) .

Humans are exposed to trans-2-hexenal daily by contamination with food and beverages. The human body was exposed to trans-2-hexenal in an amount of ~350 μg/kg/day, 98% from natural sources, and 2% from artificial flavors. Since trans-2-hexenal has a toxicity level higher than that of human normal intake by 30 times, trans-2-hexenal is unlikely to be toxic to humans. (Stout, MD; Bodes, E.; Schoonhoven, R.; Upton, PB; Travlos, GS; Swenberg, JA "Toxicity, DNA Binding, and Cell Proliferation in Male F344 Rats following Short-term Gavage Exposures to Trans-2- Hexenal." Soc. Toxicologic. Pathology March 24 2008, 1533-1601 online). In another rat study, the amount of trans-2-hexenal fed to the diet was 0, 260, 640, 1600 or 4000 ppm, and feeding for 13 weeks did not cause any changes in hematological parameters or organ weight. When the feeding amount was 4000 ppm, the body weight and intake decreased, but it was not significant (Gaunt, IF; Colley, J. “Acute and Short-term Toxicity Studies on trans-2-Hexenal.” Fd Cosmet. Toxicol. 1971 , v. 9,775-786).

Even for fruits, pears and apples, they were exposed to trans-2-hexenal (12.5 μL/L) 24 hours a day for 7 days without any appearance, color, hardness or soluble solid content. Or titratable acidity has any effect. The sensory quality of the “Golden Delicious” apples treated with untreated and trans-2-hexenal was evaluated by a trained tasting panel. No significant differences were observed. However, the odor was maintained at Bartlett. ", "Abate Fetel" and "Royal Gala" fruit are perceived (Neri, F.; Mari, M.; Menniti, A.; Brigati, S.; Bertolini, P. Control of Penicillium expansum in pears and apples by Trans-2-hexenal vapours. Postharvest Biol and Tech, 2006,41,101-108;Neri,F.;Mari,M.;Menniti,AM;Brigati,S. Activity of trans-2-hexenal against Penicillium expansum in'Conference' Pears. J. Appl Micrbiol 2006, v. 100, 1186-1193).

The concentration of 1.8 μg of trans-2-hexenal per ml of air can inhibit the germination of soybean seeds by nearly 100%. When the germinated seed is exposed to the saturated vapor of the aldehyde, the order of growth inhibition thereof is trans-2-hexenal>hexanal>trans-2-nonenal (Gardner, HW; Dornbos Jr., DL; Desjardins, AE Hexenal, trans-2-Hexenal, and trans-2-Nonenal Inhibit Soybean, Glycine max, Seed Germination. J. Agric. Food Chem, 1990, v. 38, 1316-1320).

The prior art does not suggest or observe the combined use of trans-2-hexenal and an organic acid, and synergistically improves the antimicrobial activity compared to the two components themselves. This implies a synergistic use in combination with essential oils and can act as a synergist for antibiotics.

Commercial mold inhibitors and bactericides consist of a single organic acid or a mixture of organic acids and formaldehyde. These acids are mainly propionic acid, benzoic acid, butyric acid, acetic acid and formic acid. Organic acids have become a major addition to reducing the incidence of foodborne infections. The mechanism by which short-chain fatty acids exert antimicrobial activity is that the undissociated (RCOOH = non-ionized) acids are lipid permeable, whereby they can pass through the microbial cell wall and ionize inside the more basic microorganisms (RCOOH->RCOO ^- + H ⁺ ), which in turn makes the cytoplasm unable to survive stably (Van Immerseel, F., JB Russell, MD Flythe, I. Gantois, L. Timbermont, F. Pasmans, F. Haesebrouck, and R. Ducatelle. 2006, “The use of Organic acids to combat Salmonella in poultry: a mechanistic explanation of the efficacy,” Avian Pathology, v. 35, no. 3, 182-188; Paster, N. 1979, “A commercial study of the efficiency of propionic acid and calcium propionate As fungistats in poultry feed, "Poult. Sci. v. 58,572-576).

Propionic acid is a more effective mold inhibitor than acetic acid, valeric acid, butyric acid, lactic acid and benzoic acid. The effective dose of propionic acid is between 0.05% and 0.25%. Conversely, the effective dose of other organic acids needs to be above 0.5% (Higgins C. and F. Brinkhaus, 1999, "Efficacy of several organic acids against mold," J Applied Poultry Res . v. 8, 480-487).

Feed corn is treated with a mixture of 0.5% propionic acid and 20% acetic acid, which has no adverse effects on the quality of weaned piglets (Rahneraa, S. and SMNeal, 1992, Preservation and use of chemically treated high-moisture corn by weanling). Pigs, J. Prod. Agric. v. 5, no. 4, 458-461). In broiler chickens, the addition of 0, 0.1, 0.2, 0.3 and 0.4% acetic acid in water does not affect the quality of the broiler chicks or the intestinal microbial count (Akbari, MR, H. Kermanshani and GA Kalidari, 2004, Effect of acetic acid administration in drinking water on performance growth characteristics and ileal microflora of broiler chickens," J. Sci. & Technol. Agric. & Natur. Resour. 8(3): 148).

Tannin is a naturally occurring fatty acid. It is an oily, colorless fluid that becomes solid at lower temperatures. It is weaker than butyric acid and almost insoluble in water. Tannin has been used as a non-selective herbicide. Scythe (57% tannic acid, 3% related fatty acids and 40% inert material) is a broad-spectrum post-emergence herbicide or burnt herbicide produced by Mycogen/Dow Chemical. The herbicidal mode of citric acid is due to: first, leakage of cell membrane during night and daytime; second, free radicals generated by sensitized chlorophyll detached from the thylakoid membrane during the day (B. Lederer, T. Fujimori., Y. Tsujino, K. Wakabayashi and P. Boger, 2004 "Phytotoxic activity of middle-chain fatty acids II: peroxidation and membrane effects." Pesticide Biochemistry and Physiology 80, 151-156).

Chadeganipour and Haims (2001) showed that the minimum inhibitory concentration (MIC) of medium-chain fatty acids for preventing the growth of M. gypseum was: MIC of citric acid in solid medium was 0.0 2 mg/ml, and tannic acid was 0.04 mg. /ml; In liquid medium, the MIC of citric acid is 0.075 mg/ml, and the citric acid is 0.05 mg/ml. The acids were tested individually, but not as a mixture (Chadeganipour and Haims, 2001 "Antifungal activities of pelargonic and capric acid on Microsporum gypseum" Mycoses v. 44, no 3-4, 109-112). N. Hirazawa et al. ("Antiparasitic effect of medium-chain fatty acids against ciliated Crptocaryon irritans infestation in the red sea bream Pagrus major," 2001 , Aquaculture v . 198, 219-228) found that tannic acid and fatty acids having 6 to 10 carbon atoms It can effectively control the growth of parasites to stimulate the growth of C. irritans , and C ₈ , C ₉ and C ₁₉ are the most effective. Trichoderma harzianum , a biocontrol agent for cocoa plants, was found to produce many chemicals. Tannic acid is one of them. Tannic acid is effective in controlling the germination and growth of cocoa pathogens (Aneja, M., Gianfagna, TJ, and Hebbar, KP 2005. " Trichoderma harzianum produces nonanoic acid, an inhibitor of spore germination and mycelial growth of two cacao pathogens". Physiol. Mol Plant Pathol. 67, 304-307).

Several U.S. patents disclose the use of citric acid as a fungicide and bactericide: the published U.S. Application Serial No. 2004/026, 685 discloses the disclosure of a An antifungal agent composed of an organic acid of a fatty acid. In a mixture consisting of an organic acid and a fatty acid, the organic acid acts as a potent synergist for the fatty acid as an antifungal agent. U.S. Patent 5,366,995 discloses the use of fatty acids and derivatives thereof to eliminate fungal and bacterial infections in plants and to improve the activity of fungicides and fungicides in plants. The formulation is composed of 80% citric acid or a salt thereof for controlling plant fungi. The fatty acids used are mainly C ₉ to C ₁₈ . U.S. Patent No. 5,342,630 discloses an inorganic salt for new insecticides plant comprising ₈ to C ₂₂ fatty acids which can improve the efficacy of C. One of the examples shows a powder product containing 2% citric acid, 2% citric acid, 80% talc, 10% sodium carbonate and 5% potassium carbonate. U.S. Patent 5,093,124 discloses a plant fungicide and arthropodice comprising an alpha monocarboxylic acid and a salt thereof. Preferably, the fungicide comprises a C ₉ to C ₁₀ fatty acid, which is activated partially neutralized alkali metal such as potassium. The mixture is described as consisting of 40% active ingredient dissolved in water, comprising 10% citric acid, 10% citric acid and 20% coconut fatty acid, all of which are neutralized with potassium hydroxide. U.S. Patent No. 6,596,763 discloses a method of using C fatty acids and derivatives skin infection control _{18 is} a C ₆ to. U.S. Patent Nos. 6,103,768 and 6,136,856 disclose the unique utility of fatty acids and derivatives to eliminate fungal and bacterial infections present in plants. This method cannot be prevented but is shown to be effective for infected conditions. Sharpshooter, a commercially available product containing 80% citric acid, 2% emulsifier and 18% surfactant, is shown to be effective against Penicillium and Botrytis. U.S. Patent No. 6,638,978 discloses an antimicrobial preservative, a fatty acid glycerides, fatty acid (C ₆ to C ₁₈₎ and a second fatty acid (C ₆ to C ₁₈₎ composed of a binary mixture, wherein the second fatty acid for food The antiseptic property is different from the first fatty acid. WO 01/97799 discloses the use of medium chain fatty acids as antimicrobial agents. The results show that as the pH is increased from 6.5 to 7.5, the minimum inhibitory concentration (MIC) of short chain fatty acids containing 6- to 8-carbon chains will increase.

Tannin is used as a component in food disinfecting solutions for food contact in food processing establishments. A product containing 6.49% citric acid from EcoLab is used as an active ingredient as a disinfectant for all food contact surfaces (12 CFR 178.1010 b). FDA approves tannic acid as a synthetic food flavoring agent (21 CFR 172.515), adjuvants, production aids and disinfectants for food contact (12 CFR.178.1010 b), and cleaning or auxiliary lye removal of fruits and vegetables Peel disinfectant (12 CFR 173.315). Tannins are included in the list of USDA Authorized Substances, 1990, Section 5.14, Compounds for Fruit and Vegetable Cleaning.

Emulsifiers or ethoxylated nonionic surfactants such as ethoxylated castor oil produced by the reaction of oil and ethylene oxide. Ethoxylated castor oil emulsifiers are available in a variety of chain lengths depending on the amount of ethylene oxide used in the synthesis. The molar ratio can be changed from 1 molecule of castor oil to 1 to 2000 molecules of ethylene oxide. The ethoxylated castor oil emulsifier is also named as PEG-x (polyethylene glycol) castor oil emulsifier. Wherein "x" refers to the number of molecules of ethylene oxide (Fruijtier-Polloth, Claudia, 2005, "Safety assessment on polyethylene glycols (PEGs) and their derivatives as used in cosmetic products" Toxicology, v, 214, 1-38). These emulsifiers have been widely used to dissolve poorly soluble human or animal therapeutic drugs in water. It is a volatile, stable compound that does not hydrolyze or deteriorate upon storage. Castor oil is obtained from the seeds of Ricinus communis , which is mainly composed of ricinoleic acid, isoricinoleic acid, stearic acid and dihydroxystearic acid triglyceride. Castor oil is 90% ricinoleic acid (12-hydroxyoctadecenoic acid), a non-toxic, biodegradable, and renewable resource.

Several PCT applications for the application of ethoxylated castor oil surfactants have been filed. WO 99/60865 relates to a surfactant-water emulsion to which animal feed is added before or after heat treatment. This patent relates to emulsions that help maintain or reduce water loss during heat treatment. The emulsion consists of 1-8 parts of water and 0.005-0.5 parts of surfactant. WO 97/28896 relates to the use of surfactants to promote the dispersion of molasses. WO 96/11585 relates to the use of ethoxylated castor oil in animal feed to improve the nutritional value of the feed. WO 95/28091 relates to the addition of ethoxylated castor oil to feeds to improve the availability of nutrients in conventionally dried animal feeds, thereby increasing animal growth and reducing mortality. The four patents mention the addition of an ethoxylated castor oil surfactant, preferably an emulsion, to improve the digestibility of the hydrophobic material in the animal feed and does not show any benefit to feed production or Prevention of mold growth in feed.

Anthraquinones, generally recognized as safe (GRAS) ingredients, are ubiquitous in nature and are found primarily in essential oils in plants. Their structural unit is hydrocarbon isoprene (C ₅ H ₈ ) _n . Examples of terpenoids include citral, terpene, nerol, b-ionone, geraniol, carvacrol, eugenol, carvone, terpeniol, anethole, camphor, menthol , limonene, nerolidol, phytol, phytol, carotene, squalene, thymol, tocotrienol, perillyl alcohol, borneol, myrcene, simene, terpenes, Terpenene, linalool and others. Geraniol, tocotrienol, perillyl alcohol, b-ionone and d-limonene inhibit the liver HMG-COA reductase activity, the rate-limiting step in cholesterol synthesis, and moderately lower cholesterol levels in animals ( Elson CE and SG Yu, 1994; "The Chemoprevention of Cancer by Mevalonate-Derived Constituents of Fruits and Vegetables," J. Nutr . v. 124, 607-614). D-limonene and geraniol can reduce breast tumors (Elgebede, JA, CE Elson, A. Qureshi, MA Tanner and MN Gould, 1984, "Inhibition of DMBA-Induced Mammary Cancer by Monoterpene D-limonene," Carcinogensis v. 5, No. 5, 661-664; Elgebede, JA, CE Elson, A. Qureshi, MA Tanner and MN Gould, 1986, "Regression of Rat Primary Mammary Tumors Following Dietary D-limonene," J. Nat'l Cancer Institute v.76, No. 2,323-325; Karlson, J., AK Borg, R. Unelius, MC Shoshan, N. Wilking, U. Ringborg and S. Linder, 1996, "Inhibition of Tumor Cell Growth By Monoterpenes In Vitro: Evidence of a Ras -Independent Mechanism of Action," Anticancer Drugs v. 7, no. 4, 422-429) and inhibition of growth of transplanted tumors (Yu, SG, PJ Anderson and CE Elson, 1995, "The Efficacy of B-ionone in the Chemoprevention of Rat Mammary Carcinogensis, " J. Agri. Food Chem . v. 43, 2144-2147).

It has been found that mites inhibit the growth of bacteria and fungi in vitro (Chaumont JP and D. Leger, 1992, "Campaign Against Allergic Moulds in Dwellings, Inhibitor Properties of Essential Oil Geranium "Bourbon," Citronellol, Geraniol and Citral," Ann Pharm. Fr v. 50, no. 3, 156-166), and the growth of some internal and external parasites (Hooser, SB, VR Beasly and JJ Everitt, 1986, Effects of an Insecticidal Dip Containing D-limonene in the Cat , J. Am. Vet. Med. Assoc, v. 189, no. 8, 905-908). Geraniol was found to inhibit the growth of Candida albicans and Saccharomyces cerevisiae strains by enhancing potassium leakage rate and disrupting cell membrane fluidity (Bard, M., MR Albert, N. Gupta, CJ). Guuynn and W. Stillwell, 1988, Geraniol Interferes with Membrane Functions in Strains of Candida and Saccharomyces, Lipids v. 23, no. 6, 534-538). B-ionone has been tested for its antifungal activity by inhibiting spore germination and inhibiting its growth in agar (Mikhlin ED, VP Radina, AA Dmitrossky, LP Blinkova, and LG Button, 1983, Antifungal and Antimicrobial Activity of Some Derivatives of Beta-Ionone and Vitamin A , Prikl Biokhim Mikrobiol, v, 19, 795-803; Salt, SD, S. Tuzun and J. Kuc, 1986, Effects of B-ionone and Abscisic Acid on the Growth of Tobacco and Resistance To Blue Mold, Mimicry the Effects of Stem Infection by Peronospora Tabacina, Adam Physiol. Molec. Plant Path v. 28, 287-297). Teprenone (geranylgeranylacetone) has an antibacterial effect on Helicobacter pylori (Ishii, E., 1993, Antibacterial Activity of Terprenone, a Non Water-Soluble Antiulcer Agent, Against Helicobacter Pylori, Int. J. Med'Microbiol. Virol Parasitol Infect. Dis . v.280, no. 1-2, 239-243). In vitro, 11 different terpenoid solutions are effective in inhibiting pathogenic fine growth (Kim, J., M. Marshall and C. Wei, 1995, Antibacterial Activity of Some Essential Oil Components Against Five Foodborne Pathogens, J) Agric. Food Chem . v.43, 2839-2845). Dioxa, the trichoabdal A (derived from R. Trichocarpa ), shows a very strong antibacterial effect against Helicobacter pylori (Kadota, S., P. Basnet) , E. Ishii, T. Tamura and T. Namba, 1997, Antibacterial Activity of Trichorabdal A from Rabdosia Trichocarpa Against Helicobacter Pylori, Zentralbl. Bakteriol v . 287 _{} no.} 1 63-67). Owawunmi, 1989 (Evaluation of the Antimicrobial Activity of Citral, Letters in Applied Microbiology v. 9, no. 3 105-108), showing that a growth medium containing more than 0.01% citral reduces the concentration of E. coli, which is 0.08 % has bactericidal effect. U.S. Patent No. 5,673,468 teaches a terpene formulation based on pine oil that is used as a disinfectant or antibacterial cleanser. U.S. Patent No. 5,849,956 teaches that mites found from rice have antifungal activity. U.S. Patent No. 5,939,050 teaches the use of 2 or 3 terpenoids in combination with oral hygiene antibacterial products which exhibit synergistic effects.

An object of the present invention is to improve the bactericidal effect of an organic acid in an animal feed by including at least 10% by weight of aldehyde trans-2-hexene in the feed, based on the total weight of the organic acid. aldehyde. The antimicrobial composition can be an aqueous solution comprising a mixture of an organic acid or a combination of several organic acids and aldehydes.

The composition may further comprise an ethoxylated nonionic surfactant.

The composition may further comprise an antimicrobial steroid or an essential oil.

The organic acid may be 1- to 24-carbon chain length, may be saturated, unsaturated, cyclic, and may be substituted with a functional group comprising a halogen, a hydroxyl group, an amine group, an ether or an ester.

The surfactant may be an ethoxylated castor oil surfactant having an HLB (hydrophilic lipophilic balance) from 4 to 18. It may also contain other nonionic, ionic or amphoretic surfactants or other surfactants having similar properties such as Tween.

The quinones of the composition may include allyl disulfide, thymol, citral, eugenol, carvacrol, limonene, and parsley, or mixtures thereof.

In addition to trans-2-hexenal, the composition may contain volatile aldehydes caused by the plant lipoxygenase pathway including (2E,6Z)-nonadienal, trans-2-decene Aldehydes, and other α,β-unsaturated fatty aldehydes, ie acrolein, trans-2-butenal, 2-methyl-butenal, 2-methyl-(E)-2-butenal, 2-pentenal, trans-2-hexen-1-ol, 2-methyl-2-pentenal, 2-isopropylpropenal, 2-ethyl-2-butenal, 2- Ethyl-2-hexenal, (Z)-3-hexenal, 3,7-dimethyl-6-octenal, 3,7-dimethyl-2,6-octadienal, (2E)-3,7-dimethyl-2,6-octadienal, (2Z)-3,7-dimethyl-2,6-octadienal, trans-2-nonenal , (2E, 6Z)-nonadienal, 10-undecaldehyde, 2-dodecenal, and other α,β-unsaturated fatty aldehydes having antimicrobial and flavoring properties, and their respective alcoholic and acid forms .

The mixture of the invention comprises from 1 to 90% by weight of organic acid and from 5 to 99% by weight of trans-2-hexenal.

The mixture may comprise from 0 to 90% by weight, preferably from 10 to 55% by weight, of acetic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 10 to 55% by weight, of butyric acid.

The mixture may comprise from 0 to 90% by weight, preferably from 10 to 55% by weight, of propionic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 5 to 10% by weight, of citric acid.

The mixture may comprise from 0 to 90% by weight, preferably from 10 to 40% by weight, of lactic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 10 to 55% by weight, of formic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 20 to 30% by weight, of succinic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 1 to 10% by weight, of lauric acid.

The mixture may comprise from 0 to 90% by weight, preferably from 1 to 5% by weight, of myristic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 5 to 20% by weight, of octanoic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 1 to 20% by weight, of acetopropionic acid.

The mixture may comprise from 0 to 90% by weight, preferably from 1 to 30% by weight, of a volatile alpha, beta-unsaturated fatty aldehyde.

The mixture may comprise from 0 to 50% by weight, preferably from 0.5 to 7% by weight, of hydrazine.

The mixture may comprise from 0 to 50% by weight, preferably from 0.5 to 7% by weight, of an antimicrobial steroid.

The mixture may comprise from 0 to 20% by weight, preferably from 0.5 to 10% by weight, of surfactant.

The mixture may comprise from 0.5 to 10% by weight, preferably from 1 to 5.0% by weight, of an ethoxylated castor oil surfactant having from 1 to 200 ethylene molecules.

The mixture of the present invention may comprise from 0.5 to 10% by weight, preferably from 1 to 5.0% by weight, of surfactants having similar properties to ethoxylated castor oil surfactants.

The mixture may comprise from 0 to 97% by weight, preferably from 1 to 20% by weight, of water.

The composition is effective against various fungi present in the feed and most of the feed ingredients.

The composition is effective against various bacteria present in the feed and most of the feed ingredients.

The composition is effective against various bacteria and fungi present in water.

The composition is effective against microbial damage in the production of ethanol from fermented cellulose, starch or sugar.

Another object of the present invention is to provide a method of treating an animal feed comprising: mixing an animal feed with an effective amount of an antimicrobial composition comprising from ₁ to 90% by weight, based on the total weight, of C1 to a C ₂₄ organic acid, based on the total weight, of 10 to 55% by weight of an α,β-unsaturated fatty aldehyde such as trans-2-hexenal, with the proviso that the composition of the antimicrobial composition is at least 5 A mixture of organic acid and aldehyde by weight, 0-30% by weight of total hydrazine, 0-10% by weight of total surfactant, and water.

definition

The "volume percentage" of a component is the percentage of the total volume of the formulation or composition comprising the component.

The "organic acid" in the composition may be formic acid, acetic acid, propionic acid, butyric acid, citric acid, lactic acid, and other C ₂ to C ₂₄ fatty acids or monoglycerides, diglycerides or glycerol containing C ₁ to C ₂₄ fatty acids. Triester. These fatty acids include short chain, medium chain, long chain fatty acids or short chain, medium chain, long chain triglycerides.

The α,β-unsaturated fatty aldehyde in the composition may be acrolein, trans-2-butenal, 2-methyl-butenal, 2-methyl-(E)-2-butenal, Trans-2-hexenal, 2-pentenal, trans-2-hexen-1-ol, 2-methyl-2-pentenal, 2-isopropylpropenal, 2-ethyl 2-butenal, 2-ethyl-2-hexenal, (Z)-3-hexenal, 3,7-dimethyl-6-octenal, 3,7-dimethyl- 2,6-octadienal, (2E)-3,7-dimethyl-2,6-octadienal, (2Z)-3,7-dimethyl-2,6-octadienal , trans-2-nonenal, (2E, 2Z)-nonadienal, 10-undecaldehyde, 2-dodecenal, and other α,β-unsaturated fatty aldehydes having antimicrobial and flavoring properties And its alcohol or acid form.

The "antimicrobial steroid" in the composition may include: allyl disulfide, citral, terpene, nerol, geraniol, carvacrol, eugenol, carvone, anethole, camphor, Menthol, limonene, myostatin, carotene, thymol, borneol, myrcene, terpinene, linalool or mixtures thereof. More specifically, the steroid may comprise allyl disulfide, thymol, citral, eugenol, limonene, carvacrol and carvone, or mixtures thereof.

The term "effective amount" of a compound refers to an amount that is capable of exerting the efficacy of the compound or the performance of the effective amount, such as an amount that is non-toxic but sufficient to provide the desired antimicrobial benefit. Thus, those of ordinary skill in the art can determine the appropriate effective amount using only routine experimentation.

The formulation may vary not only in the concentration of its main component (e.g., organic acid), but also in the aldehydes, hydrazines, surfactant types, and water concentrations used. The present invention can be modified in various ways by adding or removing terpenoids, types of organic acids, aldehydes, and types of surfactants from the formulation.

The term "synergistic effect" or "synergistic effect" means that when the ingredient is added as a mixture rather than being added as a single ingredient, the antiseptic effect can be enhanced.

Composition

The composition of the present invention comprises an effective amount of an organic acid of carbon chain 1-24 and an α,β-unsaturated fatty aldehyde such as trans-2-hexenal.

Antimicrobial mites, plant extracts containing essential mites or essential oils can be used like further purified mites. Terpenes are readily commercially available or are produced by various prior art techniques known in the art, such as solvent extraction or vapor extraction/distillation and chemical synthesis.

The surfactant is non-toxic and comprises an ethoxylated castor oil surfactant having from 1 to 2000, preferably from 20 to 100, polyethylene linkage.

Preferred compositions comprise from 1 to 90% by weight of an organic acid and from 1 to 30% of trans-2-hexenal, wherein the organic acid may be from 10 to 55% by weight acetic acid, from 10 to 55% by weight propionic acid, 10- 40% by weight of lactic acid, or 5.0-10% of citric acid and mixtures thereof. Preferred compositions may also comprise from 0.5 to 7% by weight of hydrazine, from 0.5 to 10% by weight of surfactant and from 1.0 to 10% by weight of water.

method

The present invention is effective against bacteria and fungi. It is applied by providing a uniform and homogeneous distribution of the mixture in the overall feed.

The invention is applicable to water.

The invention is applicable to raw materials prior to entering the mixer.

The invention can be applied to unmixed raw materials in a mixer.

The invention is applicable to the mixing process of the original ingredients.

The invention can be applied by nozzles.

One of the purposes is to control the amount of microorganisms in feed and feed ingredients. Several mixtures of organic acids, terpenes, and aldehydes produce formulations that are effective against bacteria in buffers and feeds. Another object is to formulate an antimicrobial agent with a natural compound or a compound that is safe to use. All of the chemicals used in the present invention have now been approved for use as anti-microbial, flavor enhancers and fragrances in humans.

When trans-2-hexenal is added to organic acids and terpenes, an unexpected result, a synergistic effect, is produced.

Various publications are cited throughout the application, the entire disclosure of which is hereby incorporated by reference.

Example

Examples 1 and 2

Methods: The following formulations were prepared for repeated in vitro testing. All reagents reached the highest purity and experimental grade. Acetic acid was formulated into a 56% solution. Succinic acid is diluted to 5% solution in water due to its solubility problem. Two commercially available products, a formic acid/propionic acid blend and a formaldehyde/propionic acid blend, were used as comparative tests.

A suspension of Salmonella typhimurium was added to two tubes containing 0.05% (500 ppm) of each formulation, which was vortexed, incubated at room temperature for 24 hours, and then plated onto SMA (standard) Method agar) for 48 hours, then count the number of Salmonella colonies.

Results: The table below shows that several formulations are effective in controlling the growth of Salmonella.

Results: Studies 1 and 2.

Conclusion: Each formulation can cause different responses in terms of anti-Salmonella. Formulations containing higher levels of trans-2-hexenal and lactic acid performed better than all other products except for formaldehyde and formic acid based products.

Example 3

METHODS: Six formulations were selected from previous in-vitro studies to test their effectiveness against Salmonella in feed. A formaldehyde/propionic acid mixture was used as a comparative experiment. The poultry mixed feed is supplemented with a meat and bone meal inoculum of Salmonella typhimurium. The contaminated feed is then treated with 1, 2 or 3 kg/MT of the formulations listed in the table below. After 24 hours, 10 g of the treated sample was suspended in 90 ml of Butterfield buffer. The dilution plate was plated on XLT-4 agar and incubated at 37 ° C for 48 hours, followed by Salmonella colony count. The formulations used in this experiment are shown in the table below.

*CO-60 is an ethoxylated castor oil surfactant having 60 ethylene units.

Results: The table below shows that several formulations are effective in controlling the growth of Salmonella.

Conclusion: Formulations containing trans-2-hexenal showed better anti-Salmonella effects. Formulations containing high levels of trans-2-hexenal exhibited similar effectiveness compared to formaldehyde (33% formaldehyde) and formic acid based products.

Example 4

Method: Five formulations were selected to test their effectiveness against Salmonella typhimurium. The poultry mixed feed is supplemented with a meat and bone meal inoculum of Salmonella typhimurium. The contaminated feed is then treated with 1, 2 or 3 kg/MT of the formulations listed in the table below. After 24 hours, 10 g of the treated sample was suspended in 90 ml of Butterfield buffer. The dilution plate was plated on XLT-4 agar and incubated at 37 ° C for 48 hours, followed by Salmonella colony count. Additional samples were obtained after 7 days and 14 days after the Salmonella count was completed. The formulations used are shown in the table below.

Results: The table below shows that several formulations are effective in controlling Salmonella.

Conclusion: All formulations can result in a reduction in Salmonella counts in feed. Formulations with low levels of trans-2-hexenal (<15%) are less effective than other formulations.

Example 5

Method: Formulations #17 and #18 containing trans-2-hexenal were used to compare with six other formulations containing relatively small amounts of the aldehyde but containing higher levels of lactic acid. The poultry mixed feed is supplemented with a meat and bone meal inoculum of Salmonella typhimurium. The contaminated feed is then treated with 1, 2 or 3 kg/MT of the formulations listed in the table below. After 24 hours, 10 g of the treated sample was suspended in 90 ml of Butterfield buffer. The dilution plate was plated on XLT-4 agar and incubated at 37 ° C for 48 hours, followed by Salmonella colony count. Additional samples were obtained after 7 days and 14 days after the Salmonella count was completed. The formulations used are shown in the table below.

*CO-60 is an ethoxylated castor oil surfactant having 60 ethylene units.

Results: The table below shows that several formulations are effective in controlling Salmonella.

Conclusion: When the content of trans-2-hexenal is reduced (to 5%-25%) and the lactic acid content is increased (to 26%-40%), high content of trans-2-hexene can be obtained. A similar reaction of aldehyde (30%).

Example 6

Method: This study tested four formulations that had been shown to have anti-S. typhimurium effects, a new formulation (Formulation 27), and a formaldehyde-based anti-strain from the seven formulations tested in Example 5. Microbial agent (33% formaldehyde). The poultry mixed feed is supplemented with a meat and bone meal inoculum of Salmonella typhimurium. The contaminated feed is then treated with 1, 2 or 3 kg/MT of the formulations listed in the table below. After 24 hours, 10 g of the treated sample was suspended in 90 ml of Butterfield buffer. The dilution plate was plated on XLT-4 agar and incubated at 37 ° C for 48 hours, followed by Salmonella colony count. Additional samples were obtained after 7 days, 14 days, and 21 days after the Salmonella count was completed. The formulations used are shown in the table below.

*CO-60 is an ethoxylated castor oil surfactant having 60 ethylene units.

Results: The table below shows that several formulations are effective in controlling Salmonella.

Conclusion: All formulations can achieve efficacy above 90% 2 or 3 weeks after treatment.

Example 7

This experiment was used to determine if Formulation #18 had post-treatment for residual activity. Commercially available poultry feed is ground into fine particles by a Romer mill to ensure uniform infusion of the inoculum into the feed. The feed (1 kg subsample) was transferred to 1 gallon of glass that was randomly dispensed for processing. Prior to treatment with 0, 1, 2, or 3 kg/ton of Formulation #18, the contents of a 1 gallon glass vial were added to a laboratory scale feed mixer for 1 to 2 minutes. The above operation is repeated once for each processing level. After treatment, the feed was recontaminated with 10 g of Salmonella inoculum and mixed for an additional 2 to 3 minutes. The contents of the mixer were transferred to an original 1 gallon glass bottle, sealed, and allowed to stand at room temperature (23-24 ° C) for 1 day. Feed samples (3 10 g-subsamples/treatment) were obtained 24 hours, 7 days and 14 days after re-contamination using aseptic technique. The subsample was transferred to a dilution bottle containing 100 ml of Butterfield buffer. This series of dilutions was plated onto 2 separate XLT-4 agar media. The agar medium was cultured at 37 ° C for 48 hours before the Salmonella count.

The content of three replicate samples/treated Salmonella obtained at different time intervals was averaged and reported in the following table.

Clearly, Formulation #18 remained effective for 14 days after the feed was contaminated with Salmonella.

It will be apparent to those skilled in the art of the invention that the present invention may be modified and varied without departing from the scope of the invention. The specification and the examples are intended to be illustrative and not restrictive.

Claims (12)

An antimicrobial composition comprising: 50 to 90% by weight, based on the total weight, of a C ₁ to C ₂₄ organic acid or a mixture thereof, wherein the organic acid is selected from the group consisting of acetic acid, butyric acid, propionic acid, lactic acid, and hydrazine a group consisting of acids; 5 to 30% by weight, based on the total weight, of an α,β-unsaturated fatty aldehyde, wherein the α,β-unsaturated aliphatic aldehyde is trans-2-hexenal, 2-pentyl Alkenals, or mixtures thereof; from 0 to 50% by weight, based on the total weight, of the oxime; from 0 to 20% by weight, based on the total weight of the surfactant, and water. The composition of claim 1, wherein the surfactant is an ethoxylated castor oil surfactant having an HLB (hydrophilic lipophilic balance) of from 4 to 18. The composition of claim 1, wherein the surfactant is an ethoxylated castor oil surfactant having from 1 to 200 ethylene molecules. The composition of claim 1, wherein the steroid is selected from the group consisting of allyl disulfide, citral, terpene, nerol, geraniol, carvacrol, eugenol, carvone, A group consisting of anethole, camphor, menthol, limonene, phycoerythritol, carotene, thymol, borneol, myrcene, terpenene, linalool, and mixtures thereof. The composition of claim 1, which comprises 5% by weight of citric acid, 11-25% by weight of acetic acid, 20-50% by weight of propionic acid and 5-30 weight Amount of trans-2-hexenal. The composition of claim 1, which comprises 5% by weight of citric acid, 11% by weight of acetic acid, 50% by weight of propionic acid and 25% by weight of trans-2-hexenal. A method of treating an animal feed, comprising: mixing animal feed with an effective amount of an antimicrobial composition, the composition comprising: based on the total weight, 50 to 90% by weight of a C ₁ to C ₂₄ organic acids, or mixtures thereof, Wherein the organic acid is selected from the group consisting of acetic acid, butyric acid, propionic acid, lactic acid, and citric acid; and 5 to 30% by weight, based on the total weight, of the α,β-unsaturated fatty aldehyde, wherein the α, a β-unsaturated fatty aldehyde trans-2-hexenal, 2-pentenal, or a mixture thereof; 0 to 50% by weight, based on the total weight, of hydrazine; 0 to 20% by weight, based on the total weight Surfactant, as well as water. The method of claim 7, wherein the surfactant is an ethoxylated castor oil surfactant having an HLB (hydrophilic lipophilic balance) of from 4 to 18. The method of claim 7, wherein the surfactant is an ethoxylated castor oil surfactant having from 1 to 200 ethylene molecules. The method of claim 7, wherein the steroid is selected from the group consisting of allyl disulfide, citral, terpene, nerol, geraniol, carvacrol, eugenol, carvone, and fennel. Brain, camphor, menthol, limonene, A group consisting of phycoerythritol, carotenes, thymol, borneol, myrcene, terpinene, linalool, and mixtures thereof. The method of claim 7, comprising 5 wt% of citric acid, 11-25 wt% of acetic acid, 20-50 wt% of propionic acid, and 5-30 wt% of trans-2-hexenal. . The method of claim 7, which comprises 5% by weight of citric acid, 11% by weight of acetic acid, 50% by weight of propionic acid and 25% by weight of trans-2-hexenal.