MXPA06002060A - Composition comprising a bacteriocin and an extract from a plant of the labiatae family. - Google Patents

Abstract

The present invention provides a composition comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, where (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and where when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition.

Description

COMPOSITION COMPRISING A BACTERIOCINE AND AN EXTRACT FROM A PLANT OF THE LABIATAE FAMILY The present invention relates to a composition that exhibits a microbicidal or microbiostatic action.

BACKGROUND OF THE INVENTION Bacteriocins are antimicrobial proteins or peptides that can be produced by certain bacteria, which can eliminate or inhibit the growth of closely related bacteria. Bacteriocins produced by a lactic acid bacterium are of particular importance since they have a great potential for the conservation of food and for the control of pathogens that originate in the food. (Wessels et al., 1998.) The best known bacteriocin is nisin, which is the only bacteriocin currently authorized as an additive for food. Nisin is produced by fermenting the starter bacterium culture in milk products Lactococcus lactis subsp. Lactis, and sold as the commercial extract Nisaplin® Natural Antimicrobial (Danisco). Nisin has an unusually broad antimicrobial spectrum for a bacteriocin, being active against most Gram-positive bacteria (eg species of Bacillus, Clostridium, Listeria, lactic acid bacteria). It is usually not effective against Gram-negative bacteria, yeasts or molds. Nisin is allowed as a food preservative worldwide but its levels of use and approval for food applications are strictly regulated, varying from country to country. Other bacteriocins with potential as food preservatives have been discovered, for example pediocin, (acticin, sakacin, lactococcin, enterococin, plantaricin, leucocin) These are also active, although usually with a narrower spectrum, against Gram-positive bacteria. Use in food is currently restricted to the production of the bacteriocin in situ, for example by the growth of the producer organism in the food.Antioxidants have been widely used in food products susceptible to oxidative degeneration.An antioxidant is defined by the Food and Drug Administration (21 CFR 170.3) as "a substance used to preserve food by delaying spoilage, rancidity, or discoloration due to oxidation." Plant species or extracts may be used in foods as antioxidants and to impart flavor. The advantage of these extracts is that they are perceived as natural ingredients They are compared with chemical antioxidants such as butyl hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). The plants of the Labiatae family contain various well-known herbs. It has been shown that extracts from plants have antioxidant activity and, in some cases, antimicrobial activity (Nychas &Skandamis, 2003; Smid and Gorris, 1999; Loliger, 1989). Said extracts can be essential oils and oleoresins (extracts with essential oil used in flavorings and fragrances) or "deodorized" ', extracts having a high content of phenolic diterpene and low level of flavor-inducing compounds. The essential oils are extracted by simple steam distillation of the plant material. It is reported that the most effective antioxidant compounds in rosemary and sage are carnosic acid, carnosol and rosmarinic acid (Cuvelier et al., 1996). Carnosic acid, a phenolic diterpene (C2oH2804), occurs naturally in leaves of plants of the family Labiatae, particularly rosemary and sage, but also thyme and marjoram. The dried rosemary or sage leaves contain .5-2.5% carnosic acid and 0.3-0.4% carnosol (US6231896). Carnosol is an oxidative artifact of carnosic acid (Wenkert et al., J. Org. Chem 30: 2931, 1965). The oxidation takes place in the presence of the harvest of the leaves that are left to dry in the air and if the leaves are subjected to extraction with solvents. Rosmanol can also be a product of the oxidation of carnosic acid. The use of extracts of plant material for the inhibition of the growth of micro-organisms has been taught in the art. Examples of such teachings include: WO 02/069741 which teaches extracts of Labiatae herb and hop extracts for the extension of the presence of color and the inhibition of the growth of micro-organisms in fresh meat, fish and poultry. Periago et al. 2001. Food Science &; Technology International. 7: 487-492 refers to the use of Carvacrol and thymol at 0.3 millimoles / liter in combination with nisin. It is taught that synergy was observed. JP 2001172159 relates to cosmetics comprising a range of components including antimicrobial agent and solvent extract of Labiatae. WO 98/56395 teaches a blend of tea tree oil and thyme essential oil. GB 2275 194 A discusses a disinfectant plant extract. US 6083921 discusses a combination of plant extracts including one from Labiatae: Scutellaria, preferably root (Radix scutellariae). US 5472684 teaches an oral composition for plaque and gingivitis containing thymol and eugenol. The safety of food and the prevention of food spoilage is a concern that is always present worldwide, particularly with the increasing tendency of convenient foods such as ready-to-eat foods, soups, sauces or sandwiches. Food spoilage is a major economic problem for food processing. Food manufacturers need to protect the health and safety of the public through managed products that are safe to eat. These foods can have a shelf life guaranteed, either at cold storage temperature or at room temperature. Consumers prefer high quality food with good flavor - this is difficult to achieve with chemical preservatives, irregular heating regimes and other processing measures. Food safety and protection is best achieved with a multiple preservation system using a combined method of moderate processing and natural preservatives. Micro-organisms of food origin are also less able to adapt and grow in preserved food with different conservation measures. There is great concern about the safety of food and the growth of food pathogens such as Listeria monocytogenes. This particular pathogen can grow at low temperatures, which are often used as an additional conservative measure. Pathogens that originate in food can sometimes be adapted to different preservatives and storage conditions, therefore a combination of conservative measures may be more successful than individual measures. There is an increasing need to develop economical, natural and effective food preservation systems to meet public demand for convenient, natural, safe, healthy, good quality food products with guaranteed shelf life. Bacteriocins such as nisin can be used as preservatives in food to help meet this need. Nisin is a natural conservative that has proven to be safe with GRAS status. Other bacteriocins can be used for conservation if they are produced in situ, by growth of the bacteriocin-producing organism in the feed. In some cases the required levels of bacteriocin to ensure the conservation or safety of the food may not be economically convenient, or are below effective levels due to regulatory and regulatory limitations. When bacteriocins are produced in situ, the resulting levels of bacteriocin may not be high enough to achieve the required conservative effect. The present invention solves the problems of the prior art. In one aspect the present invention provides a composition comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. In one aspect the present invention provides a method for preventing and / or inhibiting the growth of, and / or elimination of, a microorganism in a material, the method comprising the step of contacting the material with (a) an antimicrobial material.; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. In one aspect the present invention provides the use of (a) an antimicrobial material; and b) an extract obtained from or obtained from a plant of the Labiatae family, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material; where (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. In one aspect the present invention provides equipment for the preparation of a composition as defined in the present invention, the equipment comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition, in separate packages or containers; optionally with instructions for mixing and / or for putting in contact and / or for use. The aspects of the invention are defined in the appended claims. From the plant family Labiatae, rosemary and sage have antioxidant activity in foods that are mainly related to phenolic diterpenes such as carnosic acid and carnosol, as well as other phenolic compounds, including phenolic triterpenes such as betulinic acid, oleanolic acid and acid. ursolic; and rosmarinic acid. The antimicrobial activity has been attributed to some of these compounds, all of which can be obtained by selective extraction from the plants. The phenolic diterpenes, phenolic triterpenes and rosmarinic acid are different from the essential oils and oleoresins that are frequently used in flavorings and fragrances. The high levels of flavor and odor of essential oils are not conducive to their use in food. One skilled in the art could expect a combination of an antimicrobial material and an extract from the Labiatae plant family to provide a simple bactericidal or bacteriostatic additive effect. However, the in vitro studies described in the present invention have demonstrated synergistic improvement of bacteriocin activity by a deodorized extract of Rosmarinus officinaüs. This improved activity was also observed in a food model, the increasing bacteriocin (for example nisin) eliminates and controls the growth of Gram positive bacteria. The improved activity of the bacteriocin was also observed with rosemary extracts specifically prepared to contain high levels of the phenolic diterpenes camosol and carnosic acid, indicating that these compounds play an important role in the synergy. The improved activity of the bacteriocin was also observed with rosmarinic acid. The present invention provides a synergistic combination of components to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material, such as a food product. This combination of components allows lower levels of the antimicrobial material to be used to provide effective action and prevent the development of tolerance to the antimicrobial material. This is particularly important in food applications where dose reduction is desired and / or avoiding the development of tolerance for commercial and regulatory reasons. For ease of reference, these aspects and additional aspects of the present invention are now discussed under the headings of the appropriate section. However, the teachings under each section are not necessarily limited to each particular section.

Favorite aspects Antimicrobial Material In a preferred aspect the antimicrobial material is a bacteriocin. The antimicrobial material, such as a bacteriocin, can typically be selected from materials (bacteriocins) that can be used as preservatives in food. Preferably the antimicrobial material is selected from bacteriocins containing lanthionine, bacteriocins derived from Lactococcus, bacteriocins derived from Streptococcus, bacteriocins derived from Pedbcoccus, bacteriocins derived from Lactobacillus, bacteriocins derived from Carnobacterium, bacteriocins derived from Leuconostoc, bacteriocins derived from Enterococcus and mixtures from the same. Preferably the antimicrobial material is at least nisin. Preferably the antimicrobial material consists of nisin. Nisin is a bacteriocin containing lanthionine (US 5691301) derived from Lactococcus lactis subsp. Lactis (now known as Streptococcus-lactis) (US 5573801). In a preferred aspect of the present invention the bacteriocin used in the present invention is at least nisin.

As discussed in US 5573801, nisin is a polypeptide bacteriocin produced by the lactic acid bacterium, Lactococcus lactis subsp. Lactis (currently known as Streptococcus lactis Group N). Reportedly nisin is a collective name representing several closely related substances which have been designated as nisin compounds A, B, C, D and E (De Vuyst, L. and Vandamme, EJ 1994. Nisin, a lantibiotic produced by Lactococcus lactis subsp.lactis: properties, biosynthesis, fermentation and applications, in: bacteriocins of lactic acid bacteria, Microbiology, Genetics and Applications, Eds .: De Vuyst and Vandamme, Blackie Academic and Professional, London). The structure and properties of nisin are also discussed in the article by E. Lipinska, entitled "Nisin and Its Applications", The 25th Proceedings of the Easter School in Agriculture Science at the University of Nottingham, 1976, pp. 103-130 (1977), said article is incorporated herein by reference. In 1969, the FAO / WHO Joint Expert Committee on Food Additives established specifications for the purity and entity of nisin (FAO / WHO Joint Expert Committee on Food Additives, 1969. Specifications for identity and purity of some antibiotics. Series No. 430). This committee recognized nisin as a safe and legal conservator based on extensive toxicological evaluation. Nisin has the food additive number E234 and is classified as GRAS (Generally Recognized as Safe - for its acronym in English) (Food and Drug Administration 1988. Nisin preparation: Affirmation of GRAS status as a direct human ingredient. 53: 11247). The international activity unit (Ul onwards) was defined as 0.001 mg of an international reference preparation of nisin. Nisaplin® Natural Antimicrobial is the trade name for a nisin concentrate containing 1 million Ul per g, which is commercially available from Danisco. Nisin is a well-known and accepted food preservative with a long history of safe, effective use in food. There have been several reviews of nisin, for example Hurst 1981; 1983; Delves-Broughton, 1990; De Vuyst and Vandamme, 1994; Thomas et al. 2000; Thomas & Delves-Broughton, 2001). Nisin was discovered 50 years ago and the first commercial preparation, prepared in 1953, was Nisaplin®. Nisin has several characteristics that make it particularly suitable as a food preservative. Extensive toxicological evaluation has been carried out to demonstrate its safety. It is stable to heat, stable to acid and effective against a broad spectrum of Gram-positive bacteria. It is usually not effective against Gram-negative bacteria, yeasts or molds but activity against Gram-negative bacteria and yeast has been reported in the presence of chelating agents (PCT / US8902625, WO 89/12399). Nisin is an effective preservative in pasteurized foods and heat treated foods (eg processed cheese), cheese, pasteurized milk, dairy desserts, cream, mascarpone and other dairy products, puddings such as semolina, tapioca etc., pasteurized liquid egg, pasteurized potato products, soy products, fried fritters, Scottish pancakes, fried dough cakes, processed meat products, beverages, soups, sauces, ready-to-eat meals, canned foods, vegetable drinks) and foods with low acid content such as salad dressings, sauces, mayonnaise, beer, wine and other beverages. Although some loss of activity can be expected when used with processed foods, this can be improved for example by increasing the amount of nisin applied. As reported, effective levels of nisin to preserve food products have an interval of 25-500 IU / g or more. Other effective levels could be appreciated by one skilled in the art. For example, levels of 50-400 IU / g can be used. Since the discovery of the first bacteriocin, nisin, many other bacteriocins have now been found (Hoover, 993; Ray & Daeschel, 1994; Axelsen, 1998; Naidu, 2000; Ray et al. 2001; Ray & Miller, 2003). The pediocin bacteriocin, produced by Pediococcus pentosaceus, P. acidilactid, or LactobaciHus plantarum, can be used in the present invention. Similar to nisin, different structures of the pediocin have been described. Currently pediocin and other bacteriocins are allowed as food additives but their antibacterial activity can be achieved through the production of the bacteriocin in situ, as a consequence of the growth of the producer organism in the food. This is the purpose of commercial protective cultures such as HOLDBAC ™ Listeria (Danisco). Pediocin has a narrower antimicrobial spectrum compared to nisin, but there is much interest in its safety ability to eliminate, prevent or control the growth of the food pathogen Listeria monocytogenes (Ray &Miller, 2000). Other bacteriocins can be used in the present invention, including those generally named as divercin, leucocin, mesentericin, sakacin, curvacin, bavaricii, acidocin, bifidocin, carnobacteriocin, pisicocin, piscicolina, mundticina, enterocin, thermophilin, lacticin, plantarcinc, lactococcin, divercin , diplococcin, mesenterocin, leukonosin, carnosine, acidophilin, lactacin, brevicin, lactocin, helevticin, reutericin, propionicin.

Extract As discussed in the present invention when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition . In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.05% by weight based on the composition, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition.

In a preferred aspect, the composition comprises carvacrol in an amount of less than 0.05% by weight based on the composition, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less of 0.004% by weight, based on the composition. In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises carvone in an amount of less than 10% by weight based on the composition, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition. In a preferred aspect the composition comprises carvone in an amount of less than 10% by weight based on the composition, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.75% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition. In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises thymol in an amount of less than 15% by weight based on the composition, preferably less than 10% by weight, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight , based on the composition. In a preferred aspect the composition comprises thymol in an amount of less than 15% by weight based on the composition preferably less than 10% by weight, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight. % by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, preferably less than 0.004% by weight, based on the composition. In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises eugenol in an amount of less than 15% by weight based on the composition, preferably less than 10% by weight, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight , based on the composition. In a preferred aspect the composition comprises eugenol in an amount of less than 15% by weight based on the composition preferably less than 10% by weight, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight. % by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition. In a preferred aspect when the antimicrobial material consists of nlysine, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and each of carvone and thymol in amounts of less than 15% by weight based on the composition (preferably less than 10% by weight based on the composition, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight , preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition). In a preferred aspect, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and each of carvone and thymol in amounts of less than 15% by weight based on the composition (preferably less than 10% by weight). weight based on the composition, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition). In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and each of carvone, thymol and eugenol in amounts of less than 15% by weight based on the composition (preferably less than 10% by weight based on the composition, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less than 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition). In a preferred aspect, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and each of carvone, thymol and eugenol in amounts of less than 15% by weight based on the composition (preferably less than 10). % by weight based on the composition, preferably less than 7% by weight, preferably less than 5% by weight, preferably less than 2% by weight, preferably less than 1% by weight, preferably less than 0.75% by weight, preferably less 0.5% by weight, preferably less than 0.2% by weight, preferably less than 0.1% by weight, preferably less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight % by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the composition). In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises each of carvacrol and carvone in an amount of less than 1% by weight based on the extract. Preferably when the antimicrobial material consists of nisin the composition each comprises carvacrol and carvone in an amount of less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight. weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the extract. In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises each of carvacrol, carvone and thymol in amounts of less than 1% by weight based on the extract. Preferably when the antimicrobial material consists of nisin the composition each comprises carvacrol, carvone and thymol in an amount of less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02. % by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the extract. In a preferred aspect when the antimicrobial material consists of nisin, the composition comprises each of carvacrol, carvone, thymol and eugenol in amounts of less than 1% by weight based on the extract. Preferably when the antimicrobial material consists of nisin the composition each comprises carvacrol, "carvone, thymol and eugenol in an amount of less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the extract In a preferred aspect the composition comprises each of carvacrol and carvone in an amount of less than 1% by weight. weight based on the extract Preferably the composition comprises each of carvacrol and carvone in an amount of less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight , preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the extract In a preferred aspect the composition comprises each of carvacrol, arvone and thymol in amounts of less than 1% by weight based on the extract. Preferably the composition comprises each of carvacrol, carvone and thymol in an amount of less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the extract. In a preferred aspect the composition comprises each of carvacrol, carvone, thymol and eugenol in amounts of less than 1% by weight based on the extract. Preferably the composition comprises each of carvacrol, carvone, thymol and eugenol in an amount of less than 0.075% by weight, preferably less than 0.05% by weight, preferably less than 0.04% by weight, preferably less than 0.02% by weight, preferably less than 0.01% by weight, preferably less than 0.004% by weight, based on the extract. The extract used in the present invention is obtained from or obtained from a plant of the Labiatae family.

In one aspect the extract used in the present invention is obtained from a plant of the Labiatae family. It will be appreciated by one skilled in the art that by the term "extract" or "extracts" is meant any constituent of the plant which can be isolated from the entire plant. In one aspect the extract used in the present invention is obtained from a plant of the Labiatae family. It will be appreciated by one skilled in the art that an extract can be obtained from a plant or can be isolated from the plant, identified and then obtained from an alternative source, for example by chemical synthesis or enzymatic production. For example, the extract can be produced by eukaryotic or prokaryotic fermentation, by a genetic manipulation process. The present applicant has recognized that the products present in a plant of the Labiatae family can synergistically increase the activity of an antimicrobial material, preferably a bacteriocin. These products can be obtained from any source and will fall within the scope of the present invention. The invention comprises the use of a combination of a bacteriocin such as nisin and an extract of the plant family Labiatae, such as rosemary (Rosmarinus officinalis) or sage (Salvia officinalis) which together produce improved control of Gram-positive bacteria in a system of food. The extracts responsible for the synergy in the present invention preferably refer to the extracts of the plant family Labiatae which have been selectively extracted ("deodorized extracts") to increase their content of phenolic diterpene (such as carnosol and carnosic acid), content of phenolic triterpene (such as ursolic acid, betulinic acid and oleanolic acid) or rosmarinic acid content. These deodorized extracts can be distinguished by their high content of phenolic diterpene (for example greater than 3.5% by weight) and their low level (less than 1% by weight) of flavor-inducing compounds from essential vegetable oils and oleoresins which are used as flavorings or fragrances. Essential oils are typically extracted by simple steam distillation of the plant material. Essential oils comprise the various essential oils in plants that have the smell or taste of the plant from which they were extracted. Essential oils are typically terpenoids that frequently comprise monoterpenes. For example, an antioxidant of the rosemary extract type, which could be described as selectively extracted or deodorized, contains > 3.5% of phenolic diterpenes but less than 1% by weight of essential oils. A non-selective, flavoring extract contains 10-30% by weight of essential oils and a phenolic diterpene content of 2- > 3.5% by weight. An essential oil is commonly described as the volatile ether fraction obtained from a plant or plant part by a physical separation process such as distillation or chromatographic separation. Essential oils have also been described as a group of odorous principles, soluble in alcohol and to a limited extent soluble in water, consisting of mixtures of esters, aldehydes, ketones and terpenes. Essential oils are typically obtained by plants distilled with water; the oil that is separated from the distillate usually has highly characteristic odors identified with the vegetable origin. In the days of the alchemists, it was believed that the resulting mixture of organic compounds was the essence of the plant, hence the term "essential oil". In a preferred aspect the extract is a deodorized extract. Preferably the (deodorized) extract contains from 1.0 to 70% by weight of phenolic diterpenes, preferably from 3.5 to 70% by weight of phenolic diterpenes and less than 1% by weight of essential oil. In a preferred aspect the extract is selected from phenolic diterpenes, phenolic triterpenes and rosmarinic acid. In a preferred aspect the extract is or comprises a phenolic diterpene. Preferably the phenolic diterpene is selected from carosic acid, carnosol and methylcarnosic acid. Preferably the phenolic diterpene is selected from ca nosic acid and carnosol. In a preferred aspect the combined amount of phenolic diterpenes, and phenolic triterpenes and rosmarinic acid, based on the extract, is greater than 1.0% by weight. In a preferred aspect the combined amount of phenolic diterpenes, and phenolic triterpenes and rosmarinic acid, based on the composition, is greater than 1.0% by weight.

In a preferred aspect the combined amount of phenolic diterpenes, and phenolic triterpenes and rosmarinic acid, based on the extract, is greater than 3.5% by weight. In a preferred aspect the combined amount of phenolic diterpenes, and phenolic triterpenes and rosmarinic acid, based on the composition, is greater than 3.5% by weight. In a preferred aspect the amount of phenolic diterpenes, based on the extract, is greater than 1.0% by weight, for example greater than 5.0% by weight, greater than 10.0% by weight, greater than 20.0% by weight, or greater than 25.0 % in weigh. In a preferred aspect the amount of phenolic diterpenes, based on the composition, is greater than 1.0% by weight. In a preferred aspect the amount of phenolic diterpenes, based on the extract, is greater than 3.5% by weight. In a preferred aspect the amount of phenolic diterpenes, based on the composition, is greater than 3.5% by weight. In a preferred aspect the amount of phenolic diterpenes, based on the composition, is greater than 1.0% by weight, preferably greater than 2.0% by weight, preferably greater than 3.0% by weight, preferably greater than 3.5% by weight, preferably higher than 5.0% by weight, preferably greater than 10.0% by weight, preferably greater than 20.0% by weight, preferably greater than 40.0% by weight, preferably greater than 50.0% by weight.

In a preferred aspect the amount of phenolic diterpenes, based on the composition, is from 2.0 to 2.5% by weight, such as 2.3% by weight. In a preferred aspect the amount of phenolic diterpenes, based on the composition, is 4.0 to 4.5% by weight, such as 4.2% by weight. In a preferred aspect the amount of phenolic diterpenes, based on the extract, is greater than 1.0% by weight, preferably greater than 2.0% by weight, preferably greater than 3.0% by weight, preferably greater than 3.5% by weight, preferably higher than 5.0% by weight, preferably greater than 10.0% by weight, preferably greater than 20.0% by weight, preferably greater than 40.0% by weight, preferably greater than 50.0% by weight. In a highly preferred aspect the extract contains one or more phenolic triterpenes. Preferably the phenolic triterpenes are selected from betulinic acid, oleanolic acid, and ursolic acid. In a preferred aspect is or comprises a phenolic triterpene.

Preferably the phenolic triterpene is selected from betulinic acid, oleanolic acid, and ursolic acid. In a highly preferred aspect the amount of phenolic triterpenes, based on the extract, is greater than 3.5% by weight. In a highly preferred aspect the amount of phenolic triterpenes, based on the composition, is greater than 3.5% by weight.

In a preferred aspect the extract is or comprises rosmarinic acid. In a preferred aspect the amount of rosmarinic acid, based on the extract, is greater than 3.5% by weight. In a preferred aspect the amount of rosmarinic acid, based on the composition, is greater than 3.5% by weight. In a preferred aspect the extract contains flavor-inducing compounds and / or essential oils in an amount of less than 1% by weight based on the extract. In a preferred aspect the extract contains flavor-inducing compounds and / or essential oils in an amount of less than 1% by weight based on the composition. Typically the flavor-inducing compounds and / or essential oils are camphor, verbenone, borneol and alpha-terpineol. In a preferred aspect the combined amount of camphor present in the extract is less than 1% by weight (preferably less than 0.2% by weight, more preferably less than 0.15% by weight, more preferably less than 0.1% by weight) based on the abstract. In a preferred aspect the combined amount of verbenone present in the extract is less than 1% by weight (preferably less than 0.2% by weight, more preferably less than 0.15% by weight, more preferably less than 0.1% by weight) based on the abstract. In a preferred aspect the combined amount of borneol present in the extract is less than 1% by weight (preferably less than 9%). 0. 2% by weight, more preferably less than 0.15% by weight, more preferably less than 0.1% by weight) based on the extract. In a preferred aspect the combined amount of alpha-terpineol present in the extract is less than 1% by weight (preferably less than 0.2% by weight, more preferably less than 0.15% by weight, more preferably less than 0.1% by weight) based on in the extract. In a preferred aspect the combined amount of camphor, verbenone, borneol and alpha terpineol present in the extract is less than 1% by weight (preferably less than 0.2% by weight, more preferably less than 0.15% by weight, more preferably less than 0.1 % by weight) based on the extract. In a preferred aspect the extract contains less than 1% by weight of vegetable essential oils and / or oleoresins based on the extract. In a preferred aspect the extract contains less than 1% by weight of vegetable essential oils and / or oleoresins based on the composition. In a preferred aspect the extract contains essential oils in an amount of less than 1% by weight based on the extract. In a preferred aspect the extract contains essential oils in an amount of less than 1% by weight based on the composition. In a preferred aspect the plant of the Labiatae family is selected from rosemary, sage, oregano, marjoram, mint, lemon balm, savory and thyme. In a preferred aspect the plant of the Labiatae family is selected from rosemary, sage, oregano, marjoram, mint, lemon balm, and savory. It will be understood that these names cover all the species and varieties of plants known by these names. In a preferred aspect the plant of the family Labiatae is selected from rosemary (Rosmarinus officinalis L), sage. { Salvia officinalis L.) oregano. { Origanum vulgare L), marjoram. { Origanum marjorana L), mint. { Menina spp.), Lemon balm. { Melissa officinalis L), savory. { Satúrela hortensis), thyme. { Thymus vulgaris L). In a preferred aspect the plant of the Labiatae family is selected from rosemary. { Rosmarinus officinalis L), sage. { Salvia officinalis L), oregano. { Origanum vulgare L), marjoram. { Origanum marjorana L), mint. { Mentha spp.), Lemon balm. { Melissa officinalis L), and savory. { Satureia hortensis). In a preferred aspect the plant of the Labiatae family is selected from rosemary. { Rosmarinus officinalis L.), sage. { Salvia officinalis L.), marjoram. { Origanum marjorana L.), mint. { Mentha spp.), Lemon balm. { Melissa officinalis L.), and savory. { Satureia hortensis). In a preferred aspect the plant of the Labiatae family is rosemary. In a further preferred aspect phenolic diterpenes, phenolic triterpenes and rosmarinic acid are obtained by chemical synthesis. Therefore in highly preferred aspects the present invention provides a composition comprising (a) an antimicrobial material and (b) a compound selected from carnosic acid, carnosol, methicarboxylic acid, betulinic acid, oleanolic acid, ursolic acid and acid rosmarinic (preferably carnosic acid and carnosol). "A method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin; and (b) a compound selected from carnosic acid, carnosol, methiicarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid (preferably carnosic acid and carnosol). • the use of (a) an antimicrobial material and (b) a compound selected from carnosic acid, carnosol, methiicarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid (preferably carnosic acid and carnosol), to prevent and / or inhibit the growth of, and / or elimination of, a micro-organism in a material.

Microorganism As discussed herein in the present invention, it is possible to prevent and / or inhibit the growth of, and / or eliminate a micro-organism in a material. This can slow down or stop a microorganism, such as a bacterium, or eliminate the micro-organism present in contact with the present composition.

In one aspect the antimicrobial material and / or the extract are present in an amount to improve a microbicidal or microbiostatic effect. In one aspect the bacteriocin and the extract are present in an amount to provide a microbicidal or microbiostatic effect. In one aspect the bacteriocin and the extract are present in an amount to provide a microbicidal or microbiostatic synergistic effect. In one aspect the bacteriocin and the extract are present in an amount to provide a synergistic microbicidal effect. In a highly preferred aspect the microbicidal or microbiostatic effect is a bactericidal or bacteriostatic effect. It is advantageous for the bactericidal or bacteriostatic effect to be related with respect to Gram-positive bacteria and Gram-negative bacteria. Preferably the bactericidal or bacteriostatic effect is with respect to Gram positive bacteria. In a preferred aspect the bactericidal or bacteriostatic effect is with respect to an organism selected from Gram-positive bacteria associated with food spoilage or food-borne diseases including Bacillus species, Bacillus subtilis, Bacillus cereus, Listeria species, Listeria monocytogenes, lactic acid bacteria, lactic acid bacteria for food spoilage, Lactobacillus species, Staphylococcus aureus, Clostridium species, C. sporogenes, C. tyrobutyricum.

In a preferred aspect the bactericidal or bacteriostatic effect of the invention in combination with a chelating agent is with respect to an organism selected from other micro-organisms associated with spoilage of food or diseases originating in the food, including yeasts, molds and Gram-negative bacteria including Escherichia coli, Salmonella species, and Pseudomonas species. In a preferred aspect the bactericidal or bacteriostatic effect is with respect to an organism selected from Bacüíus cereus 204, B. cereus Campden, B. cereus NCTC2599, B. subtius Campden, Clostridlum sporogenes strain Campden, Clostridium sporogenes strain 1.221, Clostridium sporogenes NCIMB1793, Listeria monocytogenes 272, L monocytogenes NCTC 2426, L monocytogenes S23, Lactobacillus sake 272, Escherichia coli S15, E. coli CRA109, Salmonella Typhimurium S29, Pseudomonas fluorescens 3756. In a preferred aspect the bactericidal or bacteriostatic effect is with respect to the Listeria monocytogenes.

Food Product The composition, process and use of the present invention can prevent and / or inhibit the growth of, and / or eliminate a micro-organism in any material. However, in view of the problems associated with deterioration and contamination of food products and in view of the particular effectiveness of the present invention in food products, preferably the composition is a food product or can be added to a food product . It will be appreciated by one skilled in the art that when the present composition is a foodstuff the essential components of (a) an antimicrobial material and (b) an extract obtained from or obtained from a plant of the Labiatae family they are already present in the food product. These can be provided by one or more means. For example these may be added in the form of a composition containing the bacteriocin and the extract. The two components (the aforementioned bacteriocin and extract) can be added to the food product sequentially. In a further aspect one or more of the components can be formed in situ in the food product. For example, bacteriocin can be formed in situ in the food product by fermenting the culture of the starter bacterium in milk products Lactococcus lactis subsp. Lactis. In one aspect the composition of the present invention is a protective composition suitable for addition to a food product. Many food products can be protected by the present invention. Typical food products are raw meat, cooked meat, raw poultry products, cooked poultry products, raw fish and seafood products, cooked fish and seafood products, ready-to-eat meals, pasta sauces, pasteurized soups, mayonnaise, salad dressings, oil-in-water emulsions, margarines, low-fat spreads, water-in-oil emulsions, dairy products, cheese spreads, processed cheese, dairy desserts, flavored milk, cream, fermented milk products, cheese, butter, condensed milk products, ice cream mixes, soy products, pasteurized liquid egg, confectionery products, confectionery products, fruit products, and foods with fat-based or water-containing fillers.

Additional components The composition of the present invention or the composition for use in the present invention may contain one or more additional components. However, in certain aspects the protective composition of the present invention (suitable for addition to a food product) contains no additional components or contains no additional components that materially affect the properties of the composition. In these aspects the present invention provides a composition consisting essentially of (a) a bacteriocin and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and ( b) are different • a composition consisting of (a) a bacteriocin and (b) an extract obtained from or obtained from a plant of the Labiatae family, where (a) and (b) are different a composition consisting essentially of (a) a bacteriocin and (b) an extract obtained from or obtained from a plant of the family Labiatae, where (a) and (b) are different, wherein when the antimicrobial material consists of nisin, the composition comprising carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a composition consisting of (a) a bacteriocin and (b) an extract obtained from or obtained from a plant of the Labiatae family, where (a) and (b) are different, where when the antimicrobial material consists of nisin, the composition comprising carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. . In a preferred aspect the composition additionally comprises an emulsifier. Preferably the emulsifier is selected from polyoxyethylene sorbitan esters (E432-E436) otherwise known as polysorbates (for example Tween 80, Tween 20), monoglycerides, diglycerides, acetic acid esters of mono-diglycerides, tartaric acid mono-diglycerides and citric acid esters of mono-diglycerides. In a preferred aspect the composition additionally comprises a chelant. Preferably the chelant is selected from EDTA, citric acid, monophosphates, diphosphates, triphosphates and polyphosphates. Additional suitable chelators are taught in US 5573801 and include carboxylic acids, polycarboxylic acids, amino acids and phosphates. In particular, the following compounds and their salts may be useful: acetic acid, adenine, adipic acid, ADP, alanine, B-alanine, albumin, arginine, ascorbic acid, asparagine, aspartic acid, ATP, benzoic acid, n-butyric acid , casein, citraconic acid, citric acid, cistern, dehydracetic acid, Desferri-ferricrisina, Desferri-ferricromo, Desferri-ferrioxamina E, 3,4-Dihidroxibenzóico acid, diethylenetriaminpentaacetic acid (DTPA), Dimetilglioxima,?,? - Dimetilpurpurogalina, EDTA, formic acid, fumaric acid, globulin, gluconic acid, glutamic acid, glutaric acid, glycine, glycolic acid, glycylglycine, glycic sarcosine, guanosine, histamine, histidine, 3-hydroxy flavone, inosine, inosine triphosphate, iron free ferricrome, isovaleric acid, acid itaconic, kojic acid, lactic acid, leucine, lysine, maleic acid, malic acid, methionine, methylsalicylate, nitrilotriacetic acid (NTA), ornithine, ortho osmage, oxalic acid, oxystearin, B-phenylalanine, phosphoric acid, phytate, pimelic acid, pivalic acid, polyphosphate, proline, propionic acid, purine, pyrophosphate, pyruvic acid, riboflavin, salicylaldehyde, salicylic acid, sarcosine, serine, sorbitol, acid succinic, tartaric acid, tetrametaphosphate, thiosulfate, threonine, trimetaphosphate, triphosphate, tryptophan, uridine diphosphate, uridine triphosphate, n-valeric acid, valine, and xanthosine. Many of the aforementioned kidnapping agents are useful in food processing in their salt forms, which are comm alkaline earth metal salts or alkaline ferrous salts such as sodium, potassium or calcium or quaternary ammonium salts.

The multi-valency scavenger compounds can be used beneficially to adjust the pH or to selectively introduce or abstract metal ions for example in a food coating system. Additional information on chelators is described in T. E. Furia (Ed.), CRC Handbook of Food Additives, 2nd Ed., Pp. 271-294 (1972, Chemical Rubber Co.), and. S. Peterson and A. M. Johnson (Eds.), Encyclopaedia of Food Science, pp. 694-699 (1978, AVI Publishing Company, Inc.) such articles are incorporated herein by reference. The terms "chelant" are defined as organic or inorganic compounds capable of forming coordination complexes with metals. Also, as the term "chelator" is used in the present invention, it includes encapsulating molecular compounds such as cyclodextrin. The chelant may be inorganic or organic, but preferably is organic. Preferred chelators are non-toxic to mammals and include aminopolycarboxylic acids and their salts such as ethylenediaminetetraacetic acid (EDTA) or their salts (particularly their di and tri-sodium salts), and hydrocarboxylic acids and their salts such as citric acid. However, it is also believed that the non-citric hydrocarboxylic acid chelators and the non-citrate hydrocarboxylic acid chelators are useful in the present invention such as acetic acid, formic acid, lactic acid, tartaric acid and their salts. As mentioned above, the term "chelant" is defined and used in the present invention as a synonym for sequestering agent and is also defined to include encapsulating molecular compounds such as cyclodextrin. Cyclodextrins are cyclic carbohydrate molecules that have six, seven, or eight glucose monomers arranged in a donut-shaped ring, which are denoted alpha, beta or gamma cyclodextrin, respectively. As used in the present invention, cyclodextrin refers to both monomers and unmodified and modified cyclodextrin polymers. The molecular encapsulants of cyclodextrin are commercially available from American Maize-Products of Hammond, Ind. Cyclodextrins are further described in Chapter 11 entitled, "Industrial Applications of Cyclodextrin," by J. Szejtli, pages 331-390 of Inclusion Compounds, Vol. III (Academic Press, 1984) said chapter is incorporated herein by reference. Preferably the chelator improves the antimicrobial activity and / or antimicrobial spectrum of the bacteriocin. More preferably, the chelator improves the antimicrobial activity and / or antimicrobial spectrum of the bacteriocin with respect to Gram-negative bacteria and other microorganisms. In a preferred aspect the composition additionally comprises a lytic enzyme. Preferably the lytic enzyme is a lysozyme.

Procedure As discussed herein, in one aspect the present invention provides a method for preventing and / or inhibiting the growth of, and / or elimination of, a microorganism in a material, the method comprising the step of contacting the material with (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, where (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. In one aspect the bacteriocin and the extract are added together to the material. In one aspect the bacteriocin and the extract are added to the material sequentially. Therefore the present invention provides in one aspect a preservative / protector composition which can be added to a range of materials such as food systems and in another aspect a combination of two separate products which can be added sequentially to such materials. as food products. In one aspect the extract is added to the material.

In one aspect the bacteriocin is added to the material. In one aspect the extract is formed in situ in the material. In one aspect the bacteriocin is formed in situ in the material. Preferably when the bacteriocin is nisin, the bacteriocin can be formed in situ in the food product by fermenting the culture of the starter bacterium in milk products Lactococcus lactis subsp. Lactis.

Highly preferred aspects As discussed herein in one aspect the present invention provides a method for preventing and / or inhibiting the growth of, and / or elimination of, a microorganism in a material, the method comprising the step of contacting the material with (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. In one aspect the present invention provides the use of (a) an antimicrobial material; and b) an extract obtained from or obtained from a plant of the Labiatae family, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material; where (a) and (b) are different; and wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. Some highly preferred aspects of the present invention are set forth below: a composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) an extract obtained from a plant of the Labiatae family, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • to prevent and / or inhibit the growth of, and / or removal of a micro-organism in a material, the method comprises the step of contacting the material with (a) a bacteriocin; wherein the bacteriocin is nisin; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and where the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • the use of (a) a bacteriocin, wherein the bacteriocine is nisin; and (b) an extract obtained from or obtained from a plant of the Labiatae family, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material, wherein the The composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight. weight based on the composition • a composition comprising (a) a bacteriocin, and (b) an extract obtained from a plant of the Labiatae family selected from rosemary, sage, thyme, mint, lemon balm, savory and oregano, where (a) and (b) are different • a procedure to prevent and / or inhibit! growth of, and / or removal of a micro-organism in a material, the method comprises the step of contacting the material with (a) a bacteriocin; and (b) an extract obtained from or obtained from a plant of the Labiatae family selected from rosemary, sage, thyme, mint, lemon balm, savory and oregano, where (a) and (b) are different from • the use of (a) a bacteriocin, and (b) an extract obtained from or obtained from of a plant of the family Labiatae selected from rosemary, sage, thyme, mint, lemon balm, savory and oregano, to prevent and / or inhibit the growth of, and / or elimination of a micro-organism in a material, where (a) and (b) are different a composition comprising (a) a bacteriocin, and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid. • a method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin, and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid. • the use of (a) a bacteriocin, and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid, to prevent and / or inhibit the growth of, and / or elimination of a micro-organism in a subject! • a composition comprising (a) a bacteriocin, and (b) carnosic acid. • a method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin, and (b) carnosic acid . • the use of (a) a bacteriocin, and (b) carnosic acid, to prevent and / or inhibit the growth of, and / or elimination of, a micro-organism in a material. • a composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) an extract obtained from or obtained from a plant of the Labiatae family selected from rosemary, thyme, mint, lemon balm, savory, sage and oregano, wherein the composition contains phenolic diterpenes in a amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. "A method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin; wherein the bacteriocin is nisin; and (b) a selectively extracted extract obtained from or obtained from a plant of the Labiatae family selected from rosemary, sage, thyme, spearmint, lemon balm, savory and oregano, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based in the composition. • the use of (a) a bacteriocin, where the bacteriocin is nisin; and (b) a selectively extracted extract obtained from or obtained from a plant of the Labiatae family selected from rosemary, sage, thyme, spearmint, lemon balm, savory and oregano, to prevent and / or inhibit the growth of, and / or removal of, a microorganism in a material, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, carnosol, methylcamose acid, betulinic acid, oloanolic acid, ursolic acid and rosmarinic acid, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oloanolic acid, ursolic acid and rosmarinic acid, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. · The use of (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oloanolic acid, ursolic acid and rosmarinic acid, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. · A composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) carnosic acid. • a method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) carnosic acid. • the use of (a) a bacteriocin, where the bacteriocin is nisin; and (b) carnosic acid, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material. • a composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) an extract obtained from a plant of the Labiatae family selected from rosemary, sage, thyme, mint, lemon balm, savory and oregano, wherein the bacteriocin and the extract are present in an amount to provide an effect bactericidal or bacteriostatic synergistic, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin; wherein the bacteriocin is nisin; and (b) an extract obtained from or obtained from a plant of the family Labiatae selected from rosemary, sage, thyme, mint, lemon balm, savory and oregano, where the bacteriocin and the extract are present in an amount to provide a bactericidal or bacteriostatic synergistic effect wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight. weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • the use of (a) a bacteriocin, where the bacteriocin is nisin; and (b) an extract obtained from or obtained from a plant of the Labiatae family selected from rosemary, sage, thyme, spearmint, lemon balm, savory and oregano, to prevent and / or inhibit the growth of , and / or elimination of a micro-organism in a material, wherein the bacteriocin and the extract are present in an amount to provide a bactericidal or bacteriostatic synergistic effect, wherein the composition contains phenolic diterpenes in an amount of more than .0 % by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid, wherein the bacteriocin and the compound are present in an amount to provide a synergistic bacteriostatic or bacteriostatic effect. wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid, wherein the bacteriocin and the compound are present in an amount to provide a bactericidal or bacteriostatic synergistic effect wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the composition comprises carvacrol in an amount of less. 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. · The use of (a) a bacteriocin, where the bacteriocin is nisin; and (b) a compound selected from carnosic acid, carnosol, methylcarnosic acid, betulinic acid, oleanolic acid, ursolic acid and rosmarinic acid, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material, wherein the bacteriocin and the compound are present in an amount to provide a bactericidal or bacteriostatic synergistic effect wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein the The composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, wherein the bacteriocin and the compound are present in an amount to provide a bactericidal or bacteriostatic synergistic effect • a method for preventing and / or inhibiting the growth of, and / or elimination of a micro-organism in a material, the method comprises the step of contacting the material with (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, wherein the bacteriocin and the compound are present in an amount to provide a bactericidal or bacteriostatic synergistic effect · the use of (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) a compound selected from carnosic acid, for preventing and / or inhibiting the growth of, and / or elimination of, a microorganism in a material, wherein the bacteriocin and the compound are present in an amount to provide a bactericidal or bacteriostatic synergistic effect • a composition comprising (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) an extract obtained, from a plant of the family Labiatae, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, wherein the composition comprises carvacrol in an amount less than 0.04% by weight based on the composition and carvone in an amount of less than 0.04% by weight based on the composition. "A method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) a bacteriocin; wherein the bacteriocin is nisin; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, wherein the The composition comprises carvacrol in an amount of less than 0.04% by weight based on the composition and carvone in an amount of less than 0.04% by weight based on the composition. · The use of (a) a bacteriocin, wherein the bacteriocin is nisin; and (b) an extract obtained from or obtained from a plant of the Labiatae family, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material, wherein the The composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, wherein the composition comprises carvacrol in an amount of less than 0.04% by weight based on the composition and carvone in an amount of less than 0.04%. in weight based on the composition. The following are further defined aspects of the present invention: • a composition comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition. • a method for preventing and / or inhibiting the growth of, and / or removal of a micro-organism from a material, the method comprising the step of contacting the material with (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight. · The use of (a) an antimicrobial material; and b) an extract obtained from or obtained from a plant of the Labiatae family, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material; where (a) and (b) are different; wherein the composition contains phenoic diterpenes in an amount of more than 1.0% by weight. • equipment for the preparation of a composition as defined in the present invention, the equipment comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein the composition contains phenoic diterpenes in an amount of more than 1.0% by weight, based on the composition. • a composition comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • a method for preventing and / or inhibiting the growth of, and / or elimination of, a micro-organism in a material, the method comprising the step of contacting the material with (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • the use of (a) an antimicrobial material; and b) an extract obtained from or obtained from a plant of the Labiatae family, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material; where (a) and (b) are different; wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. • equipment for the preparation of a composition as defined in the present invention, the equipment comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different; wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition, in separate packages or containers; optionally with instructions for mixing and / or for putting in contact and / or for use. In the following, the present invention will be described in more detail by means of examples only with reference to the appended figures in which: Figure 1 is a graph showing synergistic improvement of nisin killing activity against Listeria monocytogenes in chicken soup at 25 ° C using a selectively extracted rosemary extract Figure 2 is a graph showing synergistic improvement of nisin growth control of Listeria monocytogenes in chicken soup cooled by a selectively extracted rosemary extract (GRE09) Figure 3 is a graph showing synergistic improvement by a selectively extracted rosemary extract of nisin control of the exocretion of B. cereus spore in chilled chicken soup. The minimum detection limit was 100 cfu / g. For the length of the test period, the samples containing the nisin and rosemary combination had Bacillus counts at or below 100 cfu / g. Figure 4 is a graph showing the combined effect of nisin, selectively extracted rosemary extracts and components of rosemary extract against L. monocytogenes in chicken soup at 20 ° C (minimum detection limit 00 cfu / g Figure 5 is a graph showing synergistic improvement of nisin activity by selectively extracted rosemary extracts or rosmarinic acid against Listeria monocytogenes in a chicken soup at room temperature. Figure 6 is a graph showing a demonstration of synergy between nisin and rosemary extract containing phenolic diterpene. Inhibition of L. monocytogenes at 8 ° C.

Figure 7 is a graph showing a demonstration of synergy between nisin and rosemary extract containing phenolic diterpene. Inhibition of B. cereus at 15 ° C. Figure 8 is a graph showing improved nisin growth inhibitory activity by a rosemary extract containing phenolic diterpene. Control of L. monocytogenes in carbonara sauce at 8 ° C. Figure 9 is a graph showing improved nisin growth inhibitory activity by a rosemary extract containing phenolic diterpene. Control of spores of B. cereus in carbonara sauce at 15 ° C. Figure 10 is a graph showing an improved killing effect of a nisin and rosemary extract containing phenolic diterpene against L monocytogenes in chicken soup at 20 ° C. a) pH 4.5 Figure 11 is a graph showing improved killing effect of a nisin and rosemary extract containing phenolic diterpene against L. monocytogenes in chicken soup at 20 ° C. b) pH 6.7 The present invention will be described in more detail in the following examples.

EXAMPLES Experimental evidence of benefit The in vitro studies described in the present invention have shown synergy between nisin and Rosmarinus officinalis extracts containing > 3.5% of phenolic diterpenes, significantly increasing the efficiency of nisin. This improved activity was also observed in food models, increasing elimination by nisin and controlling the growth of Gram positive bacteria. Experimental studies also showed that phenolic diterpenes, carnosic acid and carnosol were involved in this synergy. The results also indicated that rosmarinic acid can also improve the activity of nisin, through this synergistic effect that was not as strong as that observed with phenolic diterpenes.

I) In vitro demonstration of synergy between nisin and deodorised rosemary extract Materials: Rosemary Extract 09 GUARDIAN ™ (Danisco) (GRE09). This is a deodorized rosemary extract that is dispersed in water containing 4% phenolic diterpenes and < 1% of essential oils, extracted from rosemary leaves, were combined with polyoxyethylene sorbitan monooleate (Tween 80) and propylene glycol vehicles. A commercial extract of nisin at a potency of 1 x 106 IU / g: Nisaplin® Natural Antimicrobial (Danisco). Test strains: Bacillus cereus 204, B. cereus Campden, B. cereus NCTC2599, B. subtilis Campden, Listeria monocytogenes 272, L. monocytogenes NCTC12426, L. monocytogenes S23, Lactobacillus sake 272, Escherichia coli S15, E coli CRA109, Salmonella Typhimurium S29, Pseudomonas fluorescens 3756.

Method of analysis of microbial growth curve. A solution of GRE09 100,000 ppm was prepared in water and sterilized by filtration (0.2 μ ??). Additional dilutions were prepared in sterile deionized water at 1, 250-20,000 ppm. Broth was prepared from Brain Heart Infusion (Oxoid) and GRE09 storage solutions were added to produce the following GRE09 test solutions; 125, 250, 500, 750, 1000, 1250, 1500, 2000 ppm. A 10,000 U / ml nisin solution was prepared, sterilized by filtration, and then a range of storage solutions was prepared. Then a range of nisin concentrations in broth was prepared from Brain Heart Infusion. A fully automated microbial growth analyzer was used to determine the microbial growth curves (Microbiology Reader Bioscreen C analyzer associated with a PC with installed BioLink v 5.30 software, Labsystem Oy, Finland). The tests were prepared on plates for microtitre / cuvette in the form of honeycomb 2 (HC 2) with a capacity of 100 wells per plate. The wells were loaded with 270 μ? of the prepared medium and were inoculated at a level of 103 CFU (colony forming units) / ml with 30 μ? of microbial suspension. The incubation time and (at temperature were appropriate for the organism test.) This test allowed to determine suitable test levels for the compounds.Then rosemary extract and nisin in combination were evaluated, using the same procedure. Nisin were prepared at 50-1000 IU / ml in broth as mentioned above.The GRE09 solutions were prepared at 250, 500 and 1000 ppm as mentioned above.The combinations of all these test levels were prepared and evaluated in the Bioscreen as mentioned above.Results: The minimum inhibitory concentration of nisin alone, rosemary extract GRE09 alone and the two in combination on the Bioscreen after 48 hours at 30 ° C are shown in Table 1. The minimal inhibition was taken as the lowest concentration that causes the total inhibition of the bacteria after 48 hours at 30 ° C. The synergy was observed between the nisin and the rom extract. ery GRE09 against all Gram positive bacteria but no significant effect against Gram negative bacteria was observed. This can be determined from the table by comparing the data column showing MIC levels of nisin alone, GRE09 alone and the two combined. The last column gives much lower levels than the other two for the Gram positiveas bacteria (Bacillus, Listeria) but not for the Gram negative bacteria (E. coli, Salmonella).

TABLE 1 Synergy tests of nisin and rosemary extract GRE09 L. > 500 > 1000 50 + 250 50 + 500 monocytogenes 50 + 1000 S23 100 + 250 100 + 500 100 + 1000 200 + 250 200 + 500 200 + 1000 L. > 500 > 1000 50 + 250 50 + 500 monocytogenes 50 + 1000 272 100 + 250 100 + 500 100 + 1000 200 + 250 200 + 500 200 + 1000 L. > 500 > 1000 50 + 250 50 + 500 monocytogenes 50 + 1000 12426 100 + 250 100 + 500 100 + 1000 200 + 250 200 + 500 200 + 1000 E. coR S15 > 500 > 1000 > 1000 + - > 1000 E. coli CRA109 > 500 > 1000 > 1000+ > 1000 S. Typhimurium > 500 > 1000 > 1000+ -S29 > 1000 Ps. > 500 > 1000 > 1000+ Fluorescens > 1000 3756 II) Demonstration of synergy between nisin and rosemary extract GRE09 in food A) A synergy against üsteria monocyto fenes Compounds test: GRECO at 0.1%, 0.5%, Nisaplin® (Danisco). Test strains: A cocktail of L. monocytogenes strains NCTC12426, NCTC5105, NGC FS 60 and CRA3930 was prepared. The Listeria strains were grown at 30 ° C on agar with brain-heart infusion overnight and then inoculated into a broth at 30 ° C overnight. One volume of each broth was mixed together to produce a cocktail of strains with a cell concentration of approximately 09 CFU / ml. Medium: A chilled pasteurized chicken soup was used as a food model, because of this it was a good mix of different food components including vegetables, dairy products and poultry meat. This was comprised of a chicken broth with the addition of chicken, cream, vegetables, flour and seasonings. The pH was 6.12. After the addition of nisin and rosemary extract GRE09, the soup was pasteurized at a core temperature of 80 ° C for 2 minutes. The Listeria cocktail was diluted to 104 CFU / ml and inoculated into soup tests to produce a final cell count of approximately 102 CFU / g (growth inhibitory tests) and 107 CFU / ml (exterminating tests). The last test was incubated at 25 ° C for 2 hours and then evaluated by enumeration of viable count to estimate the degree of exterminating activity. The growth test was incubated at 8 ° C with regular sampling to estimate the bacteriostatic activity. Results The rosemary extract GRE09 at only 0.5% did not show listericidal activity. Nisin at 250 IU / g caused a drop of 1 log in Listeria numbers after 2 hours, but only a slight delay in growth after 24 hours (figure 1). In comparison the combination of the two products tested at these levels caused a 2-3 log drop in the Listeria numbers after 2 hours. After 24 hours the cells had not yet recovered to their initial inoculum level. This was a particularly severe test for any preservation system, since the test medium was a rich dietary model, the incubation temperature was at room temperature and the bacterial numbers were high. Therefore, any improved nisin activity was a good indication of synergy. The incubation for the bacteriostatic test was for 43 days: The results of this are shown in Figure 2 and Table 2. The synergy of nisin / rosemary was again clearly demonstrated in the food model against the Listeria cocktail. For example, Listeria growth reached 106 CFU / ml after 13 days in the presence of 100 IU / ml nisin; after 10 days in the presence of 0.1% GRE09 but only after a much longer period, 34 days, in the presence of the combination of these two ingredients. Similarly, the growth of Listeria reached 106 CFU / ml after 13 days in the presence of 100 IU / ml of nisin; after 20 days in the presence of 0.5% GRE09. The combination of the two components resulted in the absence of growth observed at the end of the test period.

TABLE 2 Summary of Lisleria growth inhibition in chilled chicken soup (the trial lasted 43 days) During the trial period (a) Nisaplin (00 Ul / ml) + GRE09 a 0. 5%, (b) Nisaplin (250 IU / ml) + GRE09 at 0.1%, and (c) Nisaplin (250 IU / ml) + GRE09 at 0.5% produced no total aerobic viable accounts above 100 cfu / g.

B) Synergy against Bacillus cereus Strains test: A cocktail of Bacillus spores was prepared as an inoculum, using Bacillus cereus strain 204, Bacillus cereus strain 199, B. cereus strain Campden, and B. cereus strain ABC 4/9.

Additions of the test compounds were made to the chicken soup, prepared as mentioned above. The soup was pasteurized to 70 ° C for 2 minutes, cooled and inoculated with approximately 103 CFU / g of a spore cocktail of Bacillus cereus. The incubation was for 56 days. The results are shown in figure 3 and are summarized in table 3. The bacteriostatic synergy between nisin and rosemary extract GRE09 was evident. By For example, food spoilage (for example 106 CFU / ml) occurred after 13 days in the presence of 25 IU / ml of nisin, and after 10 days in the presence of 300 ppm of GRE09. In the presence of both ingredients, there was no deterioration of the food at the end of the trial (56 days).

TABLE 3 Summary of test results with chilled chicken soup inoculated with Bacillus cereus spores (trial lasted 70 days) Test conditions Days until growth reached 106 CFU / ml Control 6 Nisin at 25 IU / ml 13 Rosemary Extract GRE09 at 300 ppm 10 Rosemary Extract GRE09 at 600 ppm 13 Nisin (25 IU / ml) + GRE09 at 300 ppm > 70 Nisin (25 IU / ml + GRE09 at 600 ppm> 70 C) Synergy against Clostridium sporoaenes Strains test: A cocktail of Clostridium spores was prepared as an inoculum, using Clostridium sporogenes strain Campden, Clostridium sporogenes 1221, and Clostridium sporogenes NCIMB1793. Additions of the test compounds were made to the chicken soup, prepared as mentioned above. The soup was pasteurized at 70 ° C for 2 minutes and transferred to sterile test tubes. These were inoculated with a cocktail of spores of Clostridium sporogenes to which a heat shock was applied, at a level of 2.2 x 102 CFU / g, then anaerobic conditions were created by covering the tubes with agar. The samples were incubated at 37 ° C and monitored daily for gas production (observed by extension of the gas plug and by distinctive clostridial odor). The results for an incubation period of 27 days, showing synergy, are shown in Table 4. For example, the synergy was clearly observed by the combined effect of 50 IU / ml of nisin and 300 ppm of GRE09, which prevented growth for 27 days (the length of the trial), while both individual ingredients prevented clostridial growth for 2 days (the same period of time as the control).

TABLE 4 Summary of the results of the chicken soup test inoculated with Clostridium sporogenes spores incubated at 37 ° C (the trial lasted 27 days) III) Demonstration of in vitro synergy with different extracts of deodorized, selectively extracted rosemary and rosmarinic acid The growth curves of the strains of Listeria monocytogenes and B. cereus were analyzed in laboratory medium as described above using the Bioscreen C analyzer. Minimum inhibitory concentrations (MIC) were determined for test compounds used particularly or in combination after 24 hours at 30 ° C. The results are shown in table 5. The test compounds comprised nisin (such as Nisaplin®, Danisco), GRE09 (Danisco), pure rosmarinic acid (RA; Sigma) and a range of deodorised rosemary extracts. These had been prepared by the selected extraction either with organic solvents or C02 to obtain extracts containing 28% phenolic diterpenes (28RE); Danisco) and an extract of rosemary containing 6% rosmarinic acid (6RA, Danisco). Improved nisin activity was evident with a combination of nisin combined with pure rosmarinic acid (RA, this may be partially due to low pH levels), a combination of nisin with a rosemary extract containing 6% rosmarinic acid (6RA ) and a combination of nisin with a deodorized rosemary extract containing 28% phenolic diterpenes and < 1% essential oils (28RE). The known synergy of nisin with Tween 80 was also observed. The other propylene glycol vehicle did not improve nisin activity. Synergies can be observed, as mentioned above, by comparing the MIC levels for nisin alone, the other test compound, and the two together (see table 5).

TABLE 5 MIC after growth at 30 ° C in laboratory medium Organism MIC test on Bioscreen after 24 hours at 30 ° C Individual components Combination with nisin L. monocytogenes Nisin at 1000 IU / ml - strain S23 0.1% GRE09 0.05% GRE09 + 50 IU / ml nisin 1% RA 0.25% RA + 250 IU / ml nisin 0.5% RA + 100 IU / ml of nisin 0.75% of RA + 50 IU / ml of nisin 1% of 6 RA < 0.1% of 6 RA + 250 IU / ml of nisin 0.5% of 6RA + 50 IU / ml of nisin L. monocytogenes 500 IU / ml nisin strain 272 0.25% GRE09 < 0.05% GRE09 + 50 IU / ml nisin 1% RE28 < 0.05% RE28 + 50 IU / ml nisin > 2% Tween 80 0.5% Tween 80 + 250 IU / ml nisin L monocytogenes 250 IU / ml nisin-strain NCTC12426 0.25% GRE09 < 0.05% of GR 09 + 50 IU / ml of nisin 1% of RE28 < 0.05% RE28 + 50 IU / ml nisin > 2% Tween 80 0.5% Tween 80 + 00 Ul / ml nisin B spores 500 IU / ml nisin -Cereus Campden 0.1% GRE09 0.05% GRE09 + 50 IU / ml 1% RA 0.5% RA + 250 IU / ml of nisin 0.75% of RA + 100 IU / ml of nisin 0.75% of RA + 50 IU / ml of nisin 1% of 6 RA 0.25% of 6 RA + 00 IU / ml of nisin 0.5% of 6RA + 50 IU / ml nisin IV) Demonstration of synergy for nisin activity with different components of rosemary extract deodorized in food Strains test: Listeria monocytogenes strains 272, CRA3930 and NCTC12426 The chicken soup model was used as mentioned above. The following samples were tested: GRE09, deodorized rosemary extracts containing 28% or 70% phenolic diterpenes (RE28 and RE70, Danisco), a water-soluble rosemary extract containing 6% rosmarinic acid (6RA, Danisco) and pure rosmarinic acid (RA; Sigma). Additions to the soup were made as appropriate. The soup was pasteurized (70 ° C / 2 minutes), the pH was recorded and then the soup was inoculated with a cocktail of Listeria cells prepared as described above. The tests were incubated at 20 ° C and enumeration of the viable counts was carried out after 0, 2, 4 and 24 hours at 20 ° C. Initial levels of Listeria were 1.3 x 105 CFU / ml. The test was repeated at the two nisin levels and for different periods of time. The pH of the soup without addition was pH 6.06-6.20. The addition of rosmarinic acid to 0.1% resulted in a slight drop in pH to pH 5.75. The addition of 6% RA resulted in a pH of the soup at pH 5.75-5.78. The addition of 0.5% RE28 resulted in a pH of the soup at pH 5.98. The addition of 0.5% RE70 resulted in a pH of the soup at pH 6.10. The addition of 0.5% GRE09 resulted in a pH of the soup at pH 6.02-6.09.

The results, shown in Figures 4 and 5, indicate that all deodorized extracts evaluated and rosmarinic acid contributed to the synergy with nisin in the elimination of Listeria cells. This could not be attributed to the drop in pH caused by some of the additions. The additional synergy with Tween 80 was observed in GRE09. The results indicate that the antioxidant compounds carnosol and carnosic acid, present in 28 and 70% in two of the evaluated extracts, synergistically improved the exterminating activity and the inhibitory activity of nisin growth against Listeria monocytogenes. A synergy of nisin with rosmarinic acid was evident but not as strong.

V) Demonstration of synergistic improvement of the inhibitory activity of nisin growth in different food systems using a mixture of nisin with a rosemary extract containing phenolic diterpene A) Tests with pasteurized chicken soup Method: Different additions of nisin (such as Nisaplin®, Danisco), an extract of rosemary containing 28% phenolic diterpenes (RE28), and a mixture of nisin with rosemary extract at levels of 50 IU / mg and 4.2% phenolic diterpenes were added to a commercial chicken soup that did not contain other preservatives. After the addition of the components the soup (pH 5.8) was pasteurized at a core temperature of 70 ° C for 2 minutes. The soup was cooled to room temperature and inoculated with either a cocktail of stationary phase cells of Listeria monocytogenes strains or Bacillus cereus spores. The strain cocci comprised: L monocytogenes strains NCIMB12426, strain 358, strain 272, strain CRA3930. The B. cereus cocktail comprised strains 204, 199, ABC4 / 9 and 3.046. The initial inoculum levels were approximately 102-103 CFU / g. The Bacillus tests were incubated at 15 ° C. The Listeria tests were incubated at 8 ° C. The microbiological analysis was carried out at regular intervals (agar for plate count with milk, selective agar at Listeria Oxford). Results: The results, shown as the time it took for the bacterial numbers to reach 106 CFU / g, are summarized in table 5. The total data are shown in figures 6 and 7. The results showed that the rosemary extract alone did not had activity against Bacillus, and only had a slight activity against Listeria. Rosemary extract significantly improved nisin growth inhibitory activity.

TABLE 5 Summary of results demonstrating the synergy of phenolic nisin / diterpene against Listeria and Bacillus in a pasteurized chicken soup B) Salsa tests from pasta with pasteurized meat Method: The sauce was prepared from shredded lean beef (50%), tomatoes and juice (48.9%), starch (0.5%), salt (0.4%) and sucrose (0.2%). The beef was fried for 5 minutes until it presented a brown color, then the dry ingredients were mixed followed by the tomatoes with juice. The sauce was simmered for 10 minutes and allowed to cool before mixing to a homogenous consistency. The final pH was 5.13. The additions of nisin, rosemary extract and mixtures were made. The sauce was pasteurized at a core temperature of 80 ° C for 2 minutes. A cocktail of strains of Listeria monocytogenes (as mentioned above) was inoculated after pasteurization and the tests were incubated at 8 ° C. Results: The results, shown as the time it took for the bacterial numbers to reach 106 CFU / g, are summarized in table 6. These showed that the rosemary extract alone had no activity against Bacillus, and only had a slight activity against Listeria. Rosemary extract significantly improved nisin growth inhibitory activity.

TABLE 6 Summary of results demonstrating the synergy of phenolic nisin / diterpene against Listeria in a sauce from pasteurized meat at 8 ° C C) Testing of sauce for pasta carbonara Method. A cooled commercial pasteurized sauce was used, containing cream, smoked bacon, cheese, mascarpone, butter, starch, onion, garlic mash. Protein 7 g, carbohydrate 6 g, fat 17 g. Additions of the test compounds were carried out before pasteurization (core temperature of 70 ° C for 10 minutes). The inoculations were carried out once the sauce had cooled. Samples were analyzed regularly for microbial numbers. Results: These are shown in Figures 8 and 9. As mentioned above, phenolic diterpene containing extract (8.4 ppm) synergistically enhanced nisin growth inhibitory activity against Listeria cells and Bacillus spores. The rosemary extract alone showed no activity.

VI) Demonstration of synergistic improvement of the killing activity of nisin in a food system using a mixture of nisin with rosemary extract containing phenolic diterpene Method: The diluted chicken soup (pH 6.2) was prepared as mentioned above, and It was divided into 2 batches with a batch being adjusted to pH 4.5 with HCI. The appropriate additions of nisin, rosemary extract and mixtures were carried out, then the soup was pasteurized. A cocktail of Listeria strains was inoculated to produce an initial inoculum of 105 CFU / g. Viable cells were enumerated by microbiological analysis at 0 and 2 hours. The test mixtures contained 1) 100 IU / g of nisin + 30 ppm of rosemary extract (for example 8.4 phenolic diterpenes), and 2) 150 IU / g of nisin + 45 ppm of rosemary extract (eg 12.6 phenolic diterpenes) ).

Results: The results showed that the presence of the rosemary extract containing phenolic diterpene synergistically improved the killing activity of nisin (figures 10 and 11), particularly at more acidic conditions (figure 10). The rosemary extract alone had no significant exterminating effect.

References Combination bacteriocin + rosemary JP 07-03955 & JP 3042573 (Asam Kasei K, Lion Corp) JP 3040282 (Asam Kasei KK) Nisin synergy + Tween 80 Jung, D.-S, Bodyfelt, F. W. and Daeschel,. 1992. Influence of fat and emulsifiers on the efficacy of nisin in inhibiting Listeria monocytogenes in fluid milk. Journal of Dairy Science 75: 387-393 Synergy between essential oils and nisin Pol, I. E., Krommer, J., and Smid, E. 2002. Bioenergetic consequences of nisin combined with carvacrol towards Bacillus cereus.

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Pediocin Ray, B., and Miller, K. W. 2000. Pediocin. In: Natural Food Antimicrobial Systems, ed. A. S. Naidu. Pp. 525-566. USA: CRC Press All publications mentioned in the aforementioned specification are incorporated herein by reference. Various modifications and variations of the described methods and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to said specific modalities. In fact, it is intended that various modifications of the modes described to carry out the invention, which are obvious to those skilled in chemistry, biology, food science or related fields, are within the scope of the following claims.

Claims (60)

1. NOVELTY OF THE INVENTION CLAIMS 1. - A composition comprising (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different where the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and where when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based in the composition. 2. The composition according to claim 1, further characterized in that the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition. 3. The composition according to claim 1 or 2, further characterized in that the composition comprises carvacrol in an amount of less than 0.04% by weight based on the composition. 4. The composition according to claim 1, 2 or 3, further characterized in that the composition comprises carvacrol in an amount of less than 0.02% by weight based on the composition. 5. The composition according to any of the preceding claims, further characterized in that the composition comprises carvone in an amount of less than 0.075% by weight based on the composition. 6. The composition according to any of the preceding claims, further characterized in that the composition comprises carvone in an amount of less than 0.04% by weight based on the composition. 7. The composition according to any of the preceding claims, further characterized in that the composition comprises carvone in an amount of less than 0.02% by weight based on the composition. 8. The composition according to any of the preceding claims, further characterized in that the composition comprises thymol in an amount of less than 0.1% by weight based on the composition. 9. - The composition according to any of the preceding claims, further characterized in that the composition comprises thymol in an amount of less than 0.075% by weight based on the composition. 10. The composition according to any of the preceding claims, further characterized in that the composition comprises thymus! in an amount of less than 0.05% by weight based on the composition. 11. The composition according to any of the preceding claims, further characterized in that the antimicrobial material is a bacteriocin. 12. The composition according to any of the preceding claims, further characterized in that the bacteriocin is selected from bacteriocins containing lanthionine, bacteriocins derived from Lactococcus, bacteriocins derived from Streptococcus, bacteriocins derived from Pediococcus, bacteriocins derived from Lactobacillus, bacteriocins derived from Carnobacterium, bacteriocins derived from Leuconostoc, bacteriocins derived from Enterococcus and mixtures thereof. 13. The composition according to any of the preceding claims, further characterized in that the bacteriocin is at least nisin. 14. The composition according to any of the preceding claims, further characterized in that the extract is obtained from a plant of the family Labiatae. 15. The composition according to any of the preceding claims, further characterized in that the plant of the family Labiatae is selected from rosemary, sage, oregano, marjoram, mint, lemon balm, savory, and thyme. 16. - The composition according to any of the preceding claims, further characterized in that the plant of the Labiatae family is selected from rosemary, sage, oregano, marjoram, mint, lemon balm, and savory. 17. The composition according to any of the preceding claims, further characterized in that the plant of the family Labiatae is rosemary. 18. The composition according to any of the preceding claims, further characterized in that the composition contains phenolic diterpenes in an amount greater than 2.0% by weight, based on the composition. 19. The composition according to any of the preceding claims, further characterized in that the composition contains phenolic diterpenes in an amount greater than 3.0% by weight, based on the composition. 20. The composition according to any of the preceding claims, further characterized in that the composition contains phenolic diterpenes in an amount greater than 3.5% by weight, based on the composition. 21. The composition according to any of the preceding claims, further characterized in that the phenolic diterpene is selected from carnosic acid, carnosol, methylcar- nosic acid and mixtures thereof. 22. - The composition according to any of the preceding claims, further characterized in that the extract comprises or is selected from phenolic diterpenes, phenolic triterpenes and rosmarinic acid. 23. The composition according to any of the preceding claims, further characterized in that the extract comprises a phenolic triterpene. 24. - The composition according to claim 17, further characterized in that the amount of phenolic triterpenes, based on the composition, is greater than 3.5% by weight. 25. The composition according to claim 16, 17 or 18, further characterized in that the phenolic triterpene is selected from betulinic acid, oleanolic acid, and ursolic acid. 26. The composition according to any of the preceding claims, further characterized in that the extract comprises a rosmarinic acid. 27. The composition according to claim 20, further characterized in that the amount of rosmarinic acid, based on the composition, is greater than 3.5% by weight. 28. The composition according to any of the preceding claims, further characterized in that the combined amount of phenolic diterpenes, phenolic triterpenes and rosmarinic acid, based on the composition, is greater than 3.5% by weight. 29. - The composition according to any of the preceding claims, further characterized in that the extract contains flavor-inducing compounds and / or essential oils in an amount of less than 1% by weight based on the composition. 30. The composition according to any of the preceding claims, further characterized in that the extract contains essential oils in an amount of less than 1% by weight based on the composition. 31. - The composition according to any of the preceding claims, further characterized in that the antimicrobial material and / or the extract are present in an amount to improve a microbiocidal or microbiostatic effect. 32. - The composition according to any of the preceding claims, further characterized in that the antimicrobial material and the extract are present in an amount to improve a microbicidal or microbiostatic effect. 33. - The composition according to any of the preceding claims, further characterized in that the antimicrobial material and the extract are present in an amount to provide a synergistic microbicidal or microbiostatic effect. 34. - The composition according to any of the preceding claims, further characterized in that the antimicrobial material and the extract are present in an amount to provide a synergistic microbicidal effect. 35. The composition according to any of claims 31 to 34, further characterized in that the microbicidal or microbiostatic effect is a bactericidal or bacteriostatic effect. 36. - The composition according to claim 35, further characterized in that the bactericidal or bacteriostatic effect is with respect to Gram-positive bacteria. 37. The composition according to claim 36, further characterized in that the bactericidal or bacteriostatic effect is with respect to an organism selected from the Bacülus species, Clostridium species, Listeria monocytogenes, lactic acid bacteria, and LactobacHIus species. 38. - The composition according to any of claims 35 to 37, further characterized in that the bactericidal or bacteriostatic effect is with respect to Listeria monocytogenes. 39. The composition according to any of the preceding claims, further characterized in that the composition is a food product. 40.- The composition according to any of the preceding claims, further characterized in that the composition is a protective composition suitable for addition to a food product. 41. - The composition according to claim 39 or 40, further characterized in that the food product is selected from raw meat, cooked meat, raw poultry products, cooked poultry products, raw fish and seafood products, cooked fish and seafood products, ready-to-eat meals, pasta sauces, pasteurized soups, mayonnaise, salad dressings, oil-in-water emulsions, margarines, low-fat spreads, water-in-oil emulsions, dairy products, spreads of cheese, processed cheese, dairy desserts, flavored milk, cream, fermented milk products, cheese, butter, condensed milk products, ice cream mixes, soy products, pasteurized liquid egg, confectionery products, confectionery products, fruit, and foods with fat-based fillers or that contain water. 42. - The composition according to any of the preceding claims, further characterized in that the composition further comprises an emulsifier. 43. - The composition according to claim 42, further characterized in that the emulsifier is selected from polysorbates, monoglycerides, diglycerides, acetic acid esters of mono-diglycerides, tartaric acid esters of mono-diglycerides and citric acid esters of mono-diglycerides. 44. - The composition according to any of the preceding claims, further characterized in that the composition additionally comprises a chelant. 45. The composition according to claim 44, further characterized in that the chelant is selected from EDTA, citric acid, monophosphates, diphosphates, triphosphates and polyphosphates. 46. The composition according to claim 44 or 45, further characterized in that the chelator improves the antimicrobial activity and / or antimicrobial spectrum of the antimicrobial material. 47. The composition according to claim 44, 45 or 46, further characterized in that the chelant improves the antimicrobial activity and / or antimicrobial spectrum of the antimicrobial material with respect to the Gram-negative bacteria. 48. - The composition according to any of the preceding claims, further characterized in that the composition additionally comprises a lytic enzyme. 49. The composition according to claim 48, further characterized in that the lytic enzyme is a lysozyme. 50. - The composition according to claim 1, further characterized in that it comprises (a) an antimicrobial material, wherein the antimicrobial material is nisin; and (b) an extract obtained from a plant of the Labiatae family, wherein the extract is a phenolic triterpene selected from betulinic acid, oleanolic acid or ursolic acid wherein the antimicrobial material and the extract are present in an amount to provide a bactericidal or bacteriostatic synergistic effect. 51. The composition according to claim 1, further characterized in that it comprises (a) an antimicrobial material, wherein the antimicrobial material is nisin; and (b) an extract obtained from a plant of the Labiatae family, wherein the extract is rosmarinic acid, wherein the antimicrobial material and the extract are present in an amount to provide a bactericidal or bacteriostatic synergistic effect. , 52.- A method for preventing and / or inhibiting the growth of, and / or elimination of, a microorganism in a material, the method comprising the step of contacting the material with (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different where the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, wherein when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. 53. The method according to claim 52, further characterized in that the antimicrobial material and the extract are added together to the material. 54. - The method according to claim 52, further characterized in that the antimicrobial material and the extract are added to the material sequentially. 55. - The method according to any of claims 52 to 54, further characterized in that the bacteriocin is added to the material. 56. - The method according to claim 52, further characterized in that the bacteriocin is formed in situ in the material. 57. - The method according to any of claims 52 to 56, further characterized in that the material is a food product. 58. - The method according to any of claims 52 to 57, further characterized by the characteristics according to any of claims 2 to 51. 59.- The use of (a) an antimicrobial material; and b) an extract obtained from or obtained from a plant of the Labiatae family, to prevent and / or inhibit the growth of, and / or elimination of, a microorganism in a material; where (a) and (b) are different; wherein the composition contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, and wherein when the antimicrobial material consists of nisin, the composition comprises carol in an amount of less than 0.075% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition. 60.- The use claimed in claim 59, wherein the material is a food product. 61- The use claimed in claim 59 or 60, to prevent synergistically and / or inhibit the growth of, and / or eliminate a micro-organism in a material. 62. - The use claimed in any of claims 59 to 61, which has the features of any of claims 2 to 51. 63. - A device for preparing a composition as defined in any of claims 1 to 51, the equipment comprises (a) an antimicrobial material; and (b) an extract obtained from or obtained from a plant of the Labiatae family, wherein (a) and (b) are different in separate packages or containers; optionally with instructions for mixing and / or for putting in contact and / or for use.