EP3526342A1 - Detergent composition - Google Patents

Detergent composition

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Publication number
EP3526342A1
EP3526342A1 EP18811851.7A EP18811851A EP3526342A1 EP 3526342 A1 EP3526342 A1 EP 3526342A1 EP 18811851 A EP18811851 A EP 18811851A EP 3526342 A1 EP3526342 A1 EP 3526342A1
Authority
EP
European Patent Office
Prior art keywords
bacterium
detergent composition
seq
pathogenic
composition according
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP18811851.7A
Other languages
German (de)
French (fr)
Inventor
Abdelhamid Jabrane
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ets Pollet SA
Original Assignee
Ets Pollet SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ets Pollet SA filed Critical Ets Pollet SA
Publication of EP3526342A1 publication Critical patent/EP3526342A1/en
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/381—Microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria

Definitions

  • the present invention relates to detergent compositions having the property of specifically inhibiting the growth of pathogens or undesirable ones.
  • Pathogenic bacteria are a health hazard. This danger, due to their pathogenicity, is amplified by their multi-resistance to antibiotics. In fact, the use or overuse of antibiotics has led to the emergence of multi-resistant pathogenic strains.
  • Disinfectants may present a solution but with toxicological and ecotoxicological risks that these disinfectants present.
  • Patent application WO 2014/079938 describes the addition of enzymes to detergent compositions for the treatment of biofilms resistant to conventional disinfectants and / or biocides.
  • US patent application 2008/0233093 describes the use of very particulate strains of Bacillus ssp to reduce the formation of biofilms.
  • US 2010/0028314 discloses the use of Bacillus amyloliquefaciens strain NRRL B-50154 in the treatment of biofilms and its use in several distinct embodiments, which are related to the secretion of amylase by this bacterial strain.
  • US 2010/0008893 discloses the use of Bacillus velezensis strain NRRL B-50150 in the treatment of biofilms, here also thanks to the secretion of amylase by this bacterial strain.
  • US 2011/0230345 describes the use of the strain NRRL B-50349 Bacillus amyloliquefaciens for phytosanitary use.
  • the present invention relates to a method for identifying a bacterium capable of interacting with the growth of pathogenic or undesirable microorganisms comprising the following successive steps:
  • one or more pathogenic or unwanted micro-organism (s) is taken;
  • the culture conditions for the growth of both said pathogenic or undesirable microorganism (s) and said bacterium (to be tested) are determined and / or a culture supernatant of said bacterium (to be tested) is taken;
  • a bacterium which causes a specific reduction in the growth of at least one of said pathogenic or unwanted microorganism (s).
  • the pathogen (s) is (are) selected from Gram- bacteria, Gram + bacteria, molds and yeasts.
  • At least 2, 3, 4 or 5 microorganisms are selected which are pathogenic (s) or undesirable (s).
  • the pathogenic or undesirable microorganism include Escherichia coli, Staphylococcus aureus (eg MRSA), and another pathogen selected from Pseudomonas aeruginosa, a yeast and a fungus, preferably a , 2, or each of said pathogens is (are) resistant to antibiotic treatment.
  • the growth of other pathogenic microorganisms, such as Salmonella enterica (Enterica subsp.) And Streptococcus pyogenes is also reduced by the composition of the present invention.
  • the pathogens are known to cause diseases, for example nosocomial diseases.
  • the bacterium (to be tested) is from the Bacillus amylollecaclens group.
  • At least 100 contiguous nucleotides of the genome of the bacterium has at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ. ID.NO: 4 and / or SEQ.ID.NO: 5.
  • a related aspect of the invention relates to the use of the bacterium obtainable by the above method (in particular a bacterium of the group Bacillus amylollquefaciens and / or a bacterium having at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ.ID.NO: 4 and / or SEQ.ID.NO: 5), in a detergent composition, intended preferably cleaning surfaces and / or floors and / or points of contact.
  • Such use is advantageous for communities, such as hospitals, nurseries for young children, retirement homes for the elderly or in revalidation, department stores, canteens and kitchens.
  • a liquid detergent composition of pH between 2.5 and 11.5 comprising (i) a bacterium, (ii) one or more molecule (s) having surface and / or surfactant and / or amphiphilic and (iii) a preservative for the reduction of pathogenic and / or nosocomial bacteria present on a surface and / or points of contact.
  • the bacterium is likely to be obtained by the method described above (in particular the bacteria of the Bacillus amyloliquefaciens group and / or those having at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ.ID.NO: 4 and / or SEQ.ID.NO: 5).
  • the preservative is preferably chosen from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, methyldibromoglutaronitrile, Sodium hydroxy methyl glycinate, triclosan and Sorbate (and / or sorbic acid).
  • liquid detergent composition comprising one or more molecule (s) having surface and / or surface active and / or amphiphilic properties in an amount by weight of between 1% and 30%; one or more preservative agent (s) in an amount by weight of between 0.001% and 10% and a bacterium obtainable by the method described above (in particular a bacterium of the Bacillus amyloliquefaciens group and / or a bacterium having at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ.ID.NO: 4 and / or SEQ.ID.NO : 5).
  • the preservative of the detergent composition is selected from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, quaternary ammoniums, methyldibromoglutaronitrile, Sodium hydroxy methyl glycinate, triclosan and Sorbate (and / or sorbic acid).
  • the bacterium is present at a concentration of at least 200,000 cfu / ml, preferably at least 1000000 cfu / ml, more preferably at a concentration of at least 10,000,000 cfu / ml.
  • the detergent composition further comprises one or more compound (s) selected from (mono-) nucleotides (or nucleosides), a phytohormone, mineral nitrogen and a carbon source.
  • compound (s) selected from (mono-) nucleotides (or nucleosides), a phytohormone, mineral nitrogen and a carbon source.
  • Another related aspect of the present invention is a liquid composition
  • a liquid composition comprising at least 200,000 cfu / ml, preferably at least 1000000 cfu / ml, more preferably at least 10,000,000 cfu / ml of a bacterium of which at least 100 contiguous nucleotides of its genome has at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID. NO: 1 or SEQ.ID. NO: 3 and / or SEQ.ID. NO: 4 and / or SEQ.ID. NO: 5.
  • this liquid composition further comprises a preserving agent (selected from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, quaternary ammoniums, methyldibromoglutaronitrile, sodium hydroxy methyl glycinate, triclosan and sorbate (and / or sorbic acid) and / or one or more compounds selected from nucleotides, a phytohormone, mineral nitrogen and a source of carbon.
  • a preserving agent selected from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, quaternary ammoniums, methyldibromoglutaronitrile, sodium hydroxy methyl glycinate, triclosan
  • Figure 1 depicts the interaction between the bacterium of the invention (BS) and Candida albicans (CA) in liquid culture.
  • Figure 2 depicts the interaction between the bacterium of the invention (BS) and Staphylococcus aureus (methicillin-resistant strain; MRSA) in liquid culture.
  • BS bacterium of the invention
  • MRSA methicillin-resistant strain
  • Figure 3 depicts the interaction between the bacterium of the invention (BS) and Escherichia coli (EC) in liquid culture.
  • the inventor has surprisingly noticed that the enrichment of bacteria specific to a detergent composition intended to be applied to a surface reduces the amount of pathogens on that surface without there being any biocidal or overall disinfecting effect.
  • the inventor has succeeded in introducing this bacterium into detergent compositions which, in both their concentrated and diluted forms, are compatible with the metabolism of these bacteria.
  • a first aspect of the invention is therefore a method for identifying a bacterium capable of interacting with the growth of pathogens (or harmful microorganisms) comprising the following successive stages:
  • a bacterium to be tested (identified) or a collection of bacteria to be tested (identified) is sampled (selected);
  • one or more pathogens are sampled (selected);
  • the growth of the pathogen (s) (and / or microorganism) is determined (i) in the absence of this bacterium and (ii) in the presence of this bacterium and / or the growth of the pathogen is determined in the presence culture supernatant of this bacterium;
  • a bacterium that causes a specific reduction in the growth of at least one of the pathogens.
  • the terminology “reduction (specific) of growth” means that the amount of the pathogen in condition (ii), therefore in the presence of the bacterium to be tested or the culture supernatant of the bacterium to be tested, is less than in condition (i), so in the absence of the bacterium to be tested.
  • the amount of the pathogen after culture in condition (ii) may also be less than the quantity seeded.
  • the term "specific (growth) reduction” means that the bacterium does not have a general biocidal effect.
  • the bacterium of the invention may, for example, act in competition with the pathogen (or microorganism), or produce a factor that specifically disrupts the metabolism of the pathogen (or microorganism).
  • Measure (ii) above can be done in several ways that can be cumulated. In particular, it is possible to take a supernatant from a culture of the bacteria to be tested and measure the effect of this supernatant on the growth of pathogens or the microorganism (in monoculture).
  • An alternative is to perform a co-culture test in which a pre-determined amount of the bacterium and the pathogen (or microorganism) are seeded.
  • a pre-determined amount of the bacterium and the pathogen or microorganism
  • different quantity ratios between the bacterium and the pathogen are tested.
  • the pathogen is advantageously in culture on a surface (but preferably not as a biofilm). This makes it possible to demonstrate a possible phenomenon of competition for the substrate, or even the presence of an inhibitory factor that would be produced by the bacterium to be tested (eg presence of an inhibition halo).
  • a preferred culture medium for co-cultivation includes yeast extracts, protein hydrolysates (peptone) and metabolizable sugars, e.g. glucose.
  • the pH of the medium is controlled, and maintained during the culture.
  • peptone protein hydrolysates
  • metabolizable sugars e.g. glucose.
  • the pH of the medium is controlled, and maintained during the culture.
  • the bacteria to be tested may be any bacterium that does not adversely affect the properties of the detergent composition.
  • these bacteria have no known toxicity to humans or animals and do not have a general biocidal effect.
  • These bacteria are preferably natural, that is, they are not modified by genetic engineering.
  • a preferred strain of these bacteria is to be found in the Bacillus subtilis complex.
  • Bacillus amyloliquefaciens or Bacillus velezensis More preferably, the bacteria to be tested have an element of their genome having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, (approximately) 95%, 96%, 97%, 98%, 99% or even 100% identity with at least 100 contiguous nucleotides of 16S or 23S ribosomal RNA from Bacillus amyloliquefaciens (eg ATCC strains 39374 and 39320, see SEQ.ID.Our: 1, 2) or
  • Bacillus velezensis eg strain NRRL B-41580, SEQ.ID.NO: 3
  • Bacillus velezensis eg strain NRRL B-41580, SEQ.ID.NO: 3
  • a related aspect of the present invention is that of using the bacterium described above to reduce, for example on a surface, the amount of nosocomial bacteria, yeasts and / or fungi and / or which can cause diseases and / or or unwanted.
  • yeast and / or nosocomial fungi and / or which can cause diseases we find in particular certain strains of Escherichia coli, Staphylococcus aureus, in particular meticillin-resistant strains (MRSA), Pseudomonas aeruginosa and Candida albicans or even Salmonella enterica (Enterica subsp.) and Streptococcus pyogenes.
  • MRSA meticillin-resistant strains
  • Pseudomonas aeruginosa and Candida albicans
  • Salmonella enterica Enterica subsp.
  • Streptococcus pyogenes Salmonella enterica
  • Another related aspect of the present invention is the use (or incorporation) of the bacterium identified above in detergent compositions.
  • the preferred detergent compositions are those for the treatment of surfaces, particularly surfaces of communities such as hospitals, nurseries for young children, nursing homes for elderly or revalidating, department stores (including the handles of caddies), canteens.
  • the detergent compositions are not particularly limited.
  • the detergent compositions are liquids. They preferably have pH (in general the detergent compositions contain water, which does not exclude the presence, even majority, of organic solvents) and the various compounds in concentration which are compatible with the life of the bacterium (described above) for a sufficiently long period (for example several weeks or more, in general, the compositions are stable for several months, for example 10 months or more).
  • the bacterium of the invention may be formulated in an intermediate composition, preferably a liquid, which must be mixed just before use with the detergent composition, preferably in simple proportions (eg between 1: 100 and 100: 1, preferably between 1: 10 and 10: 1).
  • the detergent composition is a liquid solution (aqueous and / or comprising one or more organic solvents) and this solution comprises (besides the bacterium described above) at least one molecule having surface properties and and / or surfactant and / or amphiphilic surfactants and soaps (preferably less than 30% by weight) and one (at least one) preservative (and / or stabilizing) agent (up to 10% by weight for total preservatives present).
  • the detergent composition is aqueous, including a composition comprising one or more organic solvents
  • its pH is set, preferably between 2.5 and 11.5, advantageously between 4 and 10, or even between 7 and 10.
  • composition further comprises, advantageously, waxes and / or polymers (preferably less than 30% by weight), bleaching agents (preferably less than 15% by weight), acids, bases (preferred) or salts (preferably a total of less than 30% by weight), as well as one or more compounds selected from the group consisting of chelants, enzymes, thickeners and corrosion inhibitors.
  • the detergent composition may optionally include one or more perfume (s) and / or one or more dyeing agent (s).
  • Another related aspect of the present invention is the bacterium identified above in a liquid medium at a concentration of at least 200,000 cfu / ml, for example at least 500,000 cfu / ml, at least 1 million cfu / ml, at least 5 million cfu / ml, at least 10 million cfu / ml, for example about 40 to about 70 million cfu / ml.
  • Cfu is the English acronym for "colony forming unit", which is the usual way of quantify the presence of a bacterium by its capacity to generate colonies, once this bacterium seeded on a suitable culture medium.
  • this liquid medium further comprises at least one molecule having surface and / or surfactant and / or amphiphilic properties such as surfactants and soaps (preferably less than 30% by weight) and a preservative (stabilizing agent). ) (up to 10% by weight, preferably up to 1% by weight).
  • a preservative stabilizing agent
  • the liquid medium advantageously further comprises one or more compounds for promoting the growth of the bacterium of the invention (once placed on a surface to be cleaned), preferably chosen from nucleotides (eg, adenine, guanine, 0.1 to 1% by weight: weight of liquid medium ) / a phytohormone, for example 3-acetic indole (from 0.01% to 0.1% by weight, liquid medium ), mineral nitrogen (eg NH 4 S0 4 : 0, 1-1% by weight liquid miiieu) and a carbon source (eg. glucose).
  • nucleotides eg, adenine, guanine, 0.1 to 1% by weight: weight of liquid medium
  • a phytohormone for example 3-acetic indole (from 0.01% to 0.1% by weight, liquid medium ), mineral nitrogen (eg NH 4 S0 4 : 0, 1-1% by weight liquid miiieu) and a carbon source (eg. glucose).
  • This compound for promoting the growth of the bacterium of the invention is particularly useful for formulations where the concentration of the bacterium of the invention is lower (eg 200,000 cfu / ml, 500,000 cfu / ml, 1000000 cfu / ml)
  • the liquid medium may be aqueous, which means that it comprises water and may also include non-aqueous solvents.
  • a preferred liquid medium comprises at least 50% by weight (organic solvent weight: weight of all solvents) of a solvent (i.e. a non-aqueous liquid at room temperature) selected from the group consisting of alcohols (eg benzyl, ethanol, isopropanol), glycol ethers (eg butyldiglycol, monopropylene glycol, dipropylene glycol), petroleum distillates (eg "white spirit", petroleum ether, paraffin oil), amino compounds (ex. monoethanolamine), ketone and aldehyde derivatives (eg methylal, dimethoxy methane).
  • a solvent i.e. a non-aqueous liquid at room temperature
  • the preservative (s) (stabilizer (s)) are present at concentrations of 10% by weight maximum, preferably between 0.001% and 10%, more preferably between 0.01% and 1%, of even more preferably, between 0.05% and 0.2% (weight of the preservative (s) present (s): weight of the composition).
  • the preservative (s) are preferably selected from the group consisting of isothiazolinone (eg 1,2-benzisothiazol-3-one; 2-methyl-2H-isothiazol-3-one; Chloro-2-methyl-isothiazolin-3 (2H) -one (CMI) 2-methylisothiazolin-3 (2H) -one (MI)), formaldehyde and its liberators (eg glutaraldehyde, 2-bromo-2-nitropropane -1,3-diol; 5-bromo-5-nitro-1,3-dioxane), preservatives (stabilizers) with an amine function (eg Diazolinidylurea, Chloroacetamide), preservatives (stabilizers) with an alcohol function (ex: Phenoxypropanol, benzyl alcohol, o-phenylphenol, phenoxyethanol), benzoate and its derivatives, parabens and derivatives, methyldi
  • Example 2 Development of a liquid medium common to the bacterium of the invention and SP.
  • the inventor has noticed that all the SP strains above (same CA, whose culture is recommended rather at pH 5, 6), but also the bacterium of the invention, are capable of growing in medium A comprising an extract yeast, peptone and glucose at pH 7 and at room temperature.
  • Example 3 Analysis of the interaction between the bacterium of the invention and SP in liquid culture.
  • the starting inoculum of the strain of the bacterium of the invention was fixed at a high concentration.
  • a colony of the bacterium of the invention is taken from an agar plate and inoculated into 10 ml of medium A in a vial.
  • the number of CFU / ml is determined after a 24-hour incubation with the particle counter.
  • the concentration of the starting inoculum of the SP strains is 5.10 2 CFU / ml.
  • several colonies are removed from the agar plate and suspended in a precise volume of medium A. This culture is then counted on the particle counter and then diluted to the desired concentration.
  • the bacterium of the invention reduced EC growth, greatly reduced AC growth and had an even stronger effect on
  • the bacterium of the invention also greatly reduced the growth in liquid medium Salmonella enterica (subs Enterica) and Streptococcus pyogenes.
  • the bacterium of the present invention did not have any significant effect in liquid culture on the growth of KP or PA, which shows that the bacterium of the invention does not have general biocidal properties, and suggests a specific effect.
  • the interaction between the bacterium of the invention and each strain of SP was studied in agar medium: firstly, the effect of the culture supernatant was tested on an agar culture of each SP; on the other hand, the effect of the bacterium of the invention on the growth of SP strains was tested by streaking or by adding colonies of the bacterium of the invention to a SP carpet.
  • CA and EC were cultured, and show carpet growth.
  • Measurement of striated interactions showed significant growth of the bacterium of the invention, and a low number of CA colonies. The effect is even more dramatic in the case of MRSA, and also observable in the case of EC.
  • the inventor also cultured the SPs on agar, and after drying, 3 colonies of the bacterium of the invention.
  • the bacterium of the invention has had a significant growth, which covers CA.
  • the inventor has observed that the bacterium of the invention, where it was seeded, caused MRSA growth inhibition halos. The effect is less pronounced for other SPs.

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Abstract

Liquid detergent composition comprising a bacterium capable of interacting with the growth of pathogenic or undesirable microorganisms, and uses of said composition.

Description

Composition détergente  Detergent composition
Domaine technique de 1 ' invention invention  Technical Field of the Invention
La présente invention a trait à des compositions détergentes ayant la propriété d'inhibition spécifique de la croissance de pathogènes ou d'indésirables. The present invention relates to detergent compositions having the property of specifically inhibiting the growth of pathogens or undesirable ones.
Arrière-plan technologique de l'invention Technological background of the invention
Les bactéries pathogènes constituent un danger pour la santé. Ce danger, dû à leur pouvoir pathogène, est amplifié par leur multi résistance aux antibiotiques. En effet, l'utilisation, voire la surutilisation, des antibiotiques a conduit à l'émergence de souches pathogènes multi résistantes. Pathogenic bacteria are a health hazard. This danger, due to their pathogenicity, is amplified by their multi-resistance to antibiotics. In fact, the use or overuse of antibiotics has led to the emergence of multi-resistant pathogenic strains.
Il y a donc un besoin pour des compositions capables de limiter l'émergence de bactéries résistantes aux antibiotiques. Les désinfectants peuvent présenter une solution mais avec des risques toxicologiques et écotoxicologiques que présentent ces désinfectants .  There is therefore a need for compositions capable of limiting the emergence of bacteria resistant to antibiotics. Disinfectants may present a solution but with toxicological and ecotoxicological risks that these disinfectants present.
La demande de brevet WO 2014/079938 décrit l'addition d'enzymes à des compositions détergentes pour le traitement des biofilms résistants aux désinfectants et/ou biocides classiques.  Patent application WO 2014/079938 describes the addition of enzymes to detergent compositions for the treatment of biofilms resistant to conventional disinfectants and / or biocides.
La demande de brevet US 2008/0233093 décrit l'utilisation de souches très particulaires de Bacillus ssp pour réduire la formation de biofilms.  US patent application 2008/0233093 describes the use of very particulate strains of Bacillus ssp to reduce the formation of biofilms.
US 2010/0028314 décrit l'utilisation de la souche NRRL B-50154 de Bacillus amyloliquefaciens dans le traitement de biofilms et son utilisation dans plusieurs modes de réalisations distincts, qui sont liés à la sécrétion d'amylase par cette souche bactérienne .  US 2010/0028314 discloses the use of Bacillus amyloliquefaciens strain NRRL B-50154 in the treatment of biofilms and its use in several distinct embodiments, which are related to the secretion of amylase by this bacterial strain.
De manière similaire, US 2010/0008893 décrit l'utilisation de la souche NRRL B-50150 de Bacillus velezensis dans le traitement de biofilms, ici aussi grâce à la sécrétion d'amylase par cette souche bactérienne. Similarly, US 2010/0008893 discloses the use of Bacillus velezensis strain NRRL B-50150 in the treatment of biofilms, here also thanks to the secretion of amylase by this bacterial strain.
Enfin, US 2011/0230345 décrit l'utilisation de la souche NRRL B-50349 de Bacillus amyloliquefaciens pour un usage phytosanitaire .  Finally, US 2011/0230345 describes the use of the strain NRRL B-50349 Bacillus amyloliquefaciens for phytosanitary use.
Aucun document de l'art antérieur ne décrit l'addition de bactéries à des compositions détergentes comprenant en outre un agent de conservation dans le but d' interagir spécifiquement avec des pathogènes. No prior art document describes the addition of bacteria to detergent compositions further comprising a preservative for the purpose of specifically interacting with pathogens.
Description résumée de l ' invention Summary description of the invention
La présente invention a trait à une méthode pour l'identification d'une bactérie susceptible d'interagir avec la croissance de micro-organismes pathogènes ou indésirables comprenant les étapes successives suivantes : The present invention relates to a method for identifying a bacterium capable of interacting with the growth of pathogenic or undesirable microorganisms comprising the following successive steps:
on prélève une bactérie à tester ;  a bacteria to be tested is taken;
on prélève un ou plusieurs micro-organisme ( s ) pathogène (s) ou indésirable ( s ) ;  one or more pathogenic or unwanted micro-organism (s) is taken;
on détermine les conditions de culture permettant la croissance à la fois dudit ou desdits micro-organismes pathogènes ou indésirables et de ladite bactérie (à tester) et/ou on prélève un surnageant de culture de ladite bactérie (à tester) ;  the culture conditions for the growth of both said pathogenic or undesirable microorganism (s) and said bacterium (to be tested) are determined and / or a culture supernatant of said bacterium (to be tested) is taken;
on détermine la croissance dudit ou desdits micro organisme (s) pathogène (s) ou indésirable ( s ) (i) en l'absence de ladite bactérie (à tester) et (ii) en présence de ladite bactérie (à tester) ou dudit surnageant de culture de ladite bactérie (à tester) ;  the growth of said microorganism (s) pathogenic (s) or undesirable (s) (i) in the absence of said bacterium (to be tested) and (ii) in the presence of said bacterium (to be tested) or said culture supernatant of said bacterium (to be tested);
on identifie une bactérie qui provoque une réduction spécifique de la croissance d'au moins un dudit ou desdits micro-organisme ( s ) pathogène (s) ou indésirable ( s ) . De manière préférée, dans la méthode, le ou les pathogène (s) est (sont) sélectionné ( s ) parmi les bactéries Gram-, les bactéries Gram+, les moisissures et les levures. a bacterium is identified which causes a specific reduction in the growth of at least one of said pathogenic or unwanted microorganism (s). Preferably, in the method, the pathogen (s) is (are) selected from Gram- bacteria, Gram + bacteria, molds and yeasts.
Avantageusement, on sélectionne au moins 2, 3, 4 ou 5 micro organisme pathogène (s) ou indésirable ( s ) .  Advantageously, at least 2, 3, 4 or 5 microorganisms are selected which are pathogenic (s) or undesirable (s).
De préférence, les micro-organisme ( s ) pathogène (s) ou indésirable ( s ) comprennent Escherichia coli, Staphyloccocus aureus (ex. MRSA) , et un autre pathogène choisi parmi Pseudomonas aeruginosa, une levure et un champignon, de préférence où un, 2, ou chacun desdits pathogènes est (sont) résistant (s) à un traitement par antibiotique. La croissance d'autres micro-organismes pathogènes, tels que Salmonella enterica (subs. Enterica) et Streptococcus pyogenes est également réduite par la composition de la présente invention. Avantageusement, les pathogènes sont connus pour causer des maladies, par exemple des maladies nosocomiales.  Preferably, the pathogenic or undesirable microorganism (s) include Escherichia coli, Staphylococcus aureus (eg MRSA), and another pathogen selected from Pseudomonas aeruginosa, a yeast and a fungus, preferably a , 2, or each of said pathogens is (are) resistant to antibiotic treatment. The growth of other pathogenic microorganisms, such as Salmonella enterica (Enterica subsp.) And Streptococcus pyogenes is also reduced by the composition of the present invention. Advantageously, the pathogens are known to cause diseases, for example nosocomial diseases.
De manière préférée, la bactérie (à tester) est du groupe Baclllus amylollquefaclens.  Preferably, the bacterium (to be tested) is from the Bacillus amylollecaclens group.
De préférence au moins 100 nucléotides contigus du génome de la bactérie (à tester) a au moins 75% d'identité avec au moins 100 nucléotides contigus de la SEQ.ID.NO :1, SEQ.ID.NO :3, SEQ.ID.NO : 4 et/ou de la SEQ.ID.NO :5.  Preferably at least 100 contiguous nucleotides of the genome of the bacterium (to be tested) has at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ. ID.NO: 4 and / or SEQ.ID.NO: 5.
Un aspect lié de l'invention a trait à l'utilisation de la bactérie susceptible d'être obtenue par la méthode ci-dessus (en particulier une bactérie du groupe Baclllus amylollquefaclens et/ou une bactérie ayant au moins 75% d'identité avec au moins 100 nucléotides contigus de la SEQ.ID.NO : 1 , SEQ.ID.NO :3, SEQ.ID.NO :4 et/ou de la SEQ.ID.NO :5), dans une composition détergente, destinée de préférence au nettoyage de surfaces et/ou des sols et/ou de points de contact .  A related aspect of the invention relates to the use of the bacterium obtainable by the above method (in particular a bacterium of the group Bacillus amylollquefaciens and / or a bacterium having at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ.ID.NO: 4 and / or SEQ.ID.NO: 5), in a detergent composition, intended preferably cleaning surfaces and / or floors and / or points of contact.
Une telle utilisation est avantageuse pour les collectivités, telles que les hôpitaux, les crèches pour enfants en bas-âge, les maisons de repos pour personnes âgées ou en revalidation, les grands magasins, les cantines et les cuisines. Such use is advantageous for communities, such as hospitals, nurseries for young children, retirement homes for the elderly or in revalidation, department stores, canteens and kitchens.
Un autre aspect associé de la présente invention est l'utilisation d'une composition détergente liquide de pH compris entre 2,5 et 11,5 comprenant (i) une bactérie, (ii) une ou plusieurs molécule (s) ayant des propriétés de surface et/ou tensio-active et/ou amphiphile et (iii) un conservateur pour la réduction des bactéries pathogènes et/ou nosocomiales présentes sur une surface et/ou points de contact.  Another related aspect of the present invention is the use of a liquid detergent composition of pH between 2.5 and 11.5 comprising (i) a bacterium, (ii) one or more molecule (s) having surface and / or surfactant and / or amphiphilic and (iii) a preservative for the reduction of pathogenic and / or nosocomial bacteria present on a surface and / or points of contact.
Pour cette utilisation, de préférence, la bactérie est susceptible d'être obtenue par la méthode décrite ci-dessus (en particulier les bactéries du groupe Bacillus amyloliquefaciens et/ou celles ayant au moins 75% d'identité avec au moins 100 nucléotides contigus de la SEQ.ID.NO :1, SEQ.ID.NO :3, SEQ.ID.NO : 4 et/ou de la SEQ.ID.NO :5).  For this use, preferably, the bacterium is likely to be obtained by the method described above (in particular the bacteria of the Bacillus amyloliquefaciens group and / or those having at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ.ID.NO: 4 and / or SEQ.ID.NO: 5).
Pour cette utilisation, de préférence, le conservateur est choisi dans le groupe constitué de l' isothiazolinone, du formaldéhyde et ses libérateurs, des conservateurs avec une fonction amine, des conservateurs avec une fonction alcool, du benzoate et de ses dérivés, des parabènes et dérivés, du methyldibromoglutaronitrile, du Sodium hydroxy methyl glycinate, du triclosan et du Sorbate (et/ou acide sorbique) . For this use, the preservative is preferably chosen from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, methyldibromoglutaronitrile, Sodium hydroxy methyl glycinate, triclosan and Sorbate (and / or sorbic acid).
Un autre aspect associé de la présente invention est une composition détergente liquide comprenant une ou plusieurs molécule (s) ayant des propriété de surface et/ou tensio-active et/ou amphiphile dans une quantité pondérale comprise entre 1% et 30% ; un ou plusieurs agent (s) conservateur ( s ) dans une quantité pondérale comprise entre 0,001% et 10% et une bactérie susceptible d'être obtenue par la méthode décrite ci-dessus (en particulier une bactérie du groupe Bacillus amyloliquefaciens et/ou une bactérie ayant au moins 75% d'identité avec au moins 100 nucléotides contigus de la SEQ.ID.NO :1, SEQ.ID.NO :3, SEQ.ID.NO : 4 et/ou de la SEQ.ID.NO :5). De préférence, le conservateur de la composition détergente est choisi dans le groupe constitué de l' isothiazolinone, du formaldéhyde et ses libérateurs, des conservateurs avec une fonction amine, des conservateurs avec une fonction alcool, du benzoate et de ses dérivés, des parabènes et dérivés, des ammoniums quaternaires, du methyldibromoglutaronitrile, du Sodium hydroxy methyl glycinate, du triclosan et du Sorbate (et/ou acide sorbique) . Another related aspect of the present invention is a liquid detergent composition comprising one or more molecule (s) having surface and / or surface active and / or amphiphilic properties in an amount by weight of between 1% and 30%; one or more preservative agent (s) in an amount by weight of between 0.001% and 10% and a bacterium obtainable by the method described above (in particular a bacterium of the Bacillus amyloliquefaciens group and / or a bacterium having at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1, SEQ.ID.NO: 3, SEQ.ID.NO: 4 and / or SEQ.ID.NO : 5). Preferably, the preservative of the detergent composition is selected from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, quaternary ammoniums, methyldibromoglutaronitrile, Sodium hydroxy methyl glycinate, triclosan and Sorbate (and / or sorbic acid).
De préférence, dans la composition détergente, la bactérie est présente à une concentration d'au moins 200000 cfu/ml, de préférence au moins 1000000 cfu/ml, de manière encore plus préférée à une concentration d'au moins 10000000 cfu/ml.  Preferably, in the detergent composition, the bacterium is present at a concentration of at least 200,000 cfu / ml, preferably at least 1000000 cfu / ml, more preferably at a concentration of at least 10,000,000 cfu / ml.
De manière avantageuse, la composition détergente comprend en outre un ou plusieurs composé (s) choisis parmi les (mono- ) nucléotides (ou des nucléosides) , une phytohormone, l'azote minéral et une source de carbone.  Advantageously, the detergent composition further comprises one or more compound (s) selected from (mono-) nucleotides (or nucleosides), a phytohormone, mineral nitrogen and a carbon source.
Un autre aspect lié de la présente invention est une composition liquide comprenant au moins 200000 cfu/ml, de préférence au moins 1000000 cfu/ml, de manière encore plus préférée au moins 10000000 cfu/ml d'une bactérie dont au moins 100 nucléotides contigus de son génome a au moins 75% d'identité avec au moins 100 nucléotides contigus de SEQ.ID. NO :1 ou de SEQ.ID. NO :3 et/ou de SEQ.ID. NO :4 et/ou de SEQ.ID. NO : 5.  Another related aspect of the present invention is a liquid composition comprising at least 200,000 cfu / ml, preferably at least 1000000 cfu / ml, more preferably at least 10,000,000 cfu / ml of a bacterium of which at least 100 contiguous nucleotides of its genome has at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID. NO: 1 or SEQ.ID. NO: 3 and / or SEQ.ID. NO: 4 and / or SEQ.ID. NO: 5.
Avantageusement, cette composition liquide comprend en outre un agent conservateur (choisi dans le groupe constitué de 1' isothiazolinone, du formaldéhyde et ses libérateurs, des conservateurs avec une fonction amine, des conservateurs avec une fonction alcool, du benzoate et de ses dérivés, des parabènes et dérivés, des ammoniums quaternaires, du methyldibromoglutaronitrile, du Sodium hydroxy methyl glycinate, du triclosan et du Sorbate (et/ou acide sorbique) ) et/ou un ou plusieurs composé (s) choisis parmi les nucléotides, une phytohormone, l'azote minéral et une source de carbone. Brève description des figures Advantageously, this liquid composition further comprises a preserving agent (selected from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, quaternary ammoniums, methyldibromoglutaronitrile, sodium hydroxy methyl glycinate, triclosan and sorbate (and / or sorbic acid) and / or one or more compounds selected from nucleotides, a phytohormone, mineral nitrogen and a source of carbon. Brief description of the figures
La figure 1 décrit l'interaction entre la bactérie de l'invention (BS) et Candida albicans (CA) en culture liquide.Figure 1 depicts the interaction between the bacterium of the invention (BS) and Candida albicans (CA) in liquid culture.
La figure 2 décrit l'interaction entre la bactérie de l'invention (BS) et Staphylococcus aureus (souche résistante à la méticilline ; MRSA) en culture liquide. Figure 2 depicts the interaction between the bacterium of the invention (BS) and Staphylococcus aureus (methicillin-resistant strain; MRSA) in liquid culture.
La figure 3 décrit l'interaction entre la bactérie de l'invention (BS) et Escherichia coli (EC) en culture liquide.  Figure 3 depicts the interaction between the bacterium of the invention (BS) and Escherichia coli (EC) in liquid culture.
Description détaillée de l'invention Detailed description of the invention
L'inventeur a remarqué, de manière surprenante que l'enrichissement de bactéries spécifiques à une composition détergente destinée à être appliquée à une surface réduisait la quantité de pathogènes à cette surface sans qu'il n'y ait un effet biocide ou désinfectant global. The inventor has surprisingly noticed that the enrichment of bacteria specific to a detergent composition intended to be applied to a surface reduces the amount of pathogens on that surface without there being any biocidal or overall disinfecting effect.
En outre, l'inventeur a réussi à introduire cette bactérie dans des compositions détergentes, qui, tant dans leur formes concentrées que diluées, sont compatibles avec le métabolisme de ces bactéries.  In addition, the inventor has succeeded in introducing this bacterium into detergent compositions which, in both their concentrated and diluted forms, are compatible with the metabolism of these bacteria.
Un premier aspect de l'invention est donc une méthode pour l'identification d'une bactérie susceptible d'interagir avec la croissance de pathogènes (ou de micro organismes nuisibles) comprenant les étapes successives suivantes : A first aspect of the invention is therefore a method for identifying a bacterium capable of interacting with the growth of pathogens (or harmful microorganisms) comprising the following successive stages:
on prélève (sélectionne) une bactérie à tester (identifier) ou une collection de bactéries à tester (identifier) ; a bacterium to be tested (identified) or a collection of bacteria to be tested (identified) is sampled (selected);
on prélève (sélectionne) un ou plusieurs pathogènes (et/ou micro organismes nuisibles) ; one or more pathogens (and / or microorganisms) are sampled (selected);
on détermine les conditions de culture permettant la croissance à la fois de ce (s) pathogène (s) (et/ou micro organisme nuisible) et de cette bactérie et/ou on prélève un surnageant de culture de cette bactérie à identifier ; the culture conditions for the growth of both this pathogen (s) (and / or microorganism) and this bacterium are determined and / or a supernatant is taken culture of this bacterium to identify;
on détermine la croissance du (des) pathogène (s) (et/ou micro organisme nuisible) (i) en l'absence de cette bactérie et (ii) en présence de cette bactérie et/ou on détermine la croissance du pathogène en présence du surnageant de culture de cette bactérie ; the growth of the pathogen (s) (and / or microorganism) is determined (i) in the absence of this bacterium and (ii) in the presence of this bacterium and / or the growth of the pathogen is determined in the presence culture supernatant of this bacterium;
on identifie une bactérie qui provoque une réduction spécifique de la croissance d'au moins un des pathogènes. a bacterium is identified that causes a specific reduction in the growth of at least one of the pathogens.
Dans le contexte de la présente invention, la terminologie « réduction (spécifique) de la croissance » signifie que la quantité du pathogène en condition (ii) , donc en présence de la bactérie à tester ou du surnageant de culture de la bactérie à tester, est moindre qu'en condition (i) , donc en l'absence de la bactérie à tester. Avantageusement, la quantité du pathogène après culture en condition (ii) peut également être inférieure à la quantité ensemencée.  In the context of the present invention, the terminology "reduction (specific) of growth" means that the amount of the pathogen in condition (ii), therefore in the presence of the bacterium to be tested or the culture supernatant of the bacterium to be tested, is less than in condition (i), so in the absence of the bacterium to be tested. Advantageously, the amount of the pathogen after culture in condition (ii) may also be less than the quantity seeded.
Dans le contexte de la présente invention, la terminologie « réduction spécifique (de la croissance) » signifie que la bactérie n'a pas un effet biocide général. La bactérie de l'invention peut, à titre d'exemple, agir par compétition avec le pathogène (ou le micro organisme nuisible) , ou produire un facteur qui perturbe spécifiquement le métabolisme du pathogène (ou du micro organisme nuisible) .  In the context of the present invention, the term "specific (growth) reduction" means that the bacterium does not have a general biocidal effect. The bacterium of the invention may, for example, act in competition with the pathogen (or microorganism), or produce a factor that specifically disrupts the metabolism of the pathogen (or microorganism).
La mesure (ii) ci-dessus peut se faire de plusieurs façons qui peuvent être cumulées. En particulier on peut prélever un surnageant d'une culture de la bactérie à tester et mesurer l'effet de ce surnageant sur la croissance de pathogènes ou du micro organisme nuisible (en monoculture) .  Measure (ii) above can be done in several ways that can be cumulated. In particular, it is possible to take a supernatant from a culture of the bacteria to be tested and measure the effect of this supernatant on the growth of pathogens or the microorganism (in monoculture).
Une alternative est la réalisation d'un test en co-culture où une quantité pré-déterminée de la bactérie et du pathogène (ou du micro organisme nuisible) sont ensemencées. De manière avantageuse, différents ratios de quantité entre la bactérie et le pathogène sont testés. Le pathogène est, de manière avantageuse, en culture sur une surface (mais de préférence pas en tant que biofilm) . Ceci permet de mettre en évidence un éventuel phénomène de compétition pour le substrat, voire même la présence d'un facteur inhibiteur qui serait produit par la bactérie à tester (ex. présence d'un halo d'inhibition). An alternative is to perform a co-culture test in which a pre-determined amount of the bacterium and the pathogen (or microorganism) are seeded. Advantageously, different quantity ratios between the bacterium and the pathogen are tested. The pathogen is advantageously in culture on a surface (but preferably not as a biofilm). This makes it possible to demonstrate a possible phenomenon of competition for the substrate, or even the presence of an inhibitory factor that would be produced by the bacterium to be tested (eg presence of an inhibition halo).
Cependant, un test en co-culture en milieu liquide est possible. De manière avantageuse, les deux tests, sur surface et en milieu liquide, sont pratiqués.  However, a co-culture test in a liquid medium is possible. Advantageously, both tests on surface and in liquid medium are practiced.
Dans le cas de co-culture en milieu liquide, il est avantageux de mesurer la quantité optimale de bactérie à ensemencer, ou le ratio (quantité de bactérie à ensemencer : quantité de pathogène) optimal ou, au minimum, de tester 3, 4, 5, voire 10 ratios différents, présentant une gamme de variation d'un facteur de 10 au moins (par exemple 1:1 et 20:1) .  In the case of co-culture in a liquid medium, it is advantageous to measure the optimal amount of bacterium to be seeded, or the ratio (quantity of bacterium to be sown: amount of pathogen) optimal or, at least, to test 3, 4, 5 or 10 different ratios having a variation range of at least a factor of 10 (eg 1: 1 and 20: 1).
Un milieu de culture préféré pour la co-culture comprend des extraits de levure, des hydrolysats de protéines (peptone) et de sucres métabolisables , par exemple le glucose. De préférence, le pH du milieu est contrôlé, et maintenu au cours de la culture. L'homme du métier sait comment déterminer le pH optimum, que ce soit pour la culture simple, et pour la co culture .  A preferred culture medium for co-cultivation includes yeast extracts, protein hydrolysates (peptone) and metabolizable sugars, e.g. glucose. Preferably, the pH of the medium is controlled, and maintained during the culture. One skilled in the art knows how to determine the optimum pH, both for simple cultivation and for cultivation.
Dans le contexte de la présente invention, la bactérie à tester peut être n' importe quelle bactérie qui ne pénaliserait pas les propriétés de la composition détergente. Ainsi, ces bactéries n'ont pas de toxicité connue pour l'homme ou l'animal et n'ont pas d'effet biocide général. Ces bactéries sont, de préférence naturelles, c'est-à-dire qu'elles ne sont pas modifiées par ingénierie génétique.  In the context of the present invention, the bacteria to be tested may be any bacterium that does not adversely affect the properties of the detergent composition. Thus, these bacteria have no known toxicity to humans or animals and do not have a general biocidal effect. These bacteria are preferably natural, that is, they are not modified by genetic engineering.
Une souche préférée de ces bactéries est à trouver dans le complexe Bacillus subtilis . De préférence des Bacillus amyloliquefaciens ou Bacillus velezensis . De manière encore préférée, les bactéries à tester ont un élément de leur génome ayant au moins 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, (environ) 95%, 96%, 97%, 98%, 99% voire même ont 100% d'identité avec au moins 100 nucléotides contigus de l'ARN ribosomial 16S ou 23S de Bacillus amyloliquefaciens (ex. les souches ATCC 39374 et 39320 ; voir SEQ.ID.Nos : 1, 2) ou deA preferred strain of these bacteria is to be found in the Bacillus subtilis complex. Preferably Bacillus amyloliquefaciens or Bacillus velezensis. More preferably, the bacteria to be tested have an element of their genome having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, (approximately) 95%, 96%, 97%, 98%, 99% or even 100% identity with at least 100 contiguous nucleotides of 16S or 23S ribosomal RNA from Bacillus amyloliquefaciens (eg ATCC strains 39374 and 39320, see SEQ.ID.Our: 1, 2) or
Bacillus velezensis (ex. la souche NRRL B-41580 ; SEQ.ID.NO :3), ou encore ayant au moins 70%, 75%, 80%, 85%,Bacillus velezensis (eg strain NRRL B-41580, SEQ.ID.NO: 3), or at least 70%, 75%, 80%, 85%,
90%, 91%, 92%, 93%, 94%, (environ) 95%, 96%, 97%, 98%, 99% voire même ont 100% d'identité avec au moins 100 nucléotides contigus de l'ADN de la sous unité alpha de la Gyrase (voir SEQ.ID.NO :4, Bacillus amyloliquefaciens souche CP 1604 et/ou SEQ.ID.NO :5, Bacillus velezensis, souche B-41580). Une telle comparaison de séquence peut se faire avantageusement en utilisant BLASTn, bien connu de l'homme du métier. 90%, 91%, 92%, 93%, 94%, (approximately) 95%, 96%, 97%, 98%, 99% or even 100% identity with at least 100 contiguous nucleotides of DNA of the alpha subunit of Gyrase (see SEQ.ID.NO: 4, Bacillus amyloliquefaciens strain CP 1604 and / or SEQ.ID.NO: 5, Bacillus velezensis, strain B-41580). Such a sequence comparison can be done advantageously using BLASTn, well known to those skilled in the art.
Un aspect connexe de la présente invention est celui de l'utilisation de la bactérie décrite ci-dessus pour réduire, par exemple sur une surface, la quantité des bactéries, levures et/ou champignons nosocomiaux et/ou qui peuvent causer des maladies et/ou indésirables. A related aspect of the present invention is that of using the bacterium described above to reduce, for example on a surface, the amount of nosocomial bacteria, yeasts and / or fungi and / or which can cause diseases and / or or unwanted.
Ceux-ci sont avantageusement choisis parmi le groupe constitué de Actinomyces, Bacillus anthracis, Brucella , Campylobacter, Clostridium sp . (C. botuminium, C. perfrinqens, C. tetanl) , These are advantageously chosen from the group consisting of Actinomyces, Bacillus anthracis, Brucella, Campylobacter, Clostridium sp. (C. botuminium, C. perfrinqens, C. tetanl),
Borrelia sp., Corynebacterium sp . , Enterobacter sp . , Escherichia coli , Flavobacterium meninqosepticum, Haemophilus sp . , Hélicobacter pylori , Klebsiella sp . , Leqionella sp . , Listeria ivanovii, Listeria monocytoqenes, Mycobacterium sp . , neisseria meninqitidis, Pasteurella sp . , Proteus mirabilis, Pseudomonas aeruqinosa , Salmonella sp (ex. S. typhimurium) , Shiqella dysenteriae, Staphylococcus aureus, Streptococcus sp . (ex. S. pneumoniae ou S. pyoqenes) Vibrio sp (ex. V. cholerae) , Yersinia sp (ex. Y. pestis et Y. enterolitica) , Asperqillus fumiqatus, Candida albicans, Trichphyton sp, Bacillus cereus, Cladosporidium sp . , Pénicillium sp . , Asperqillus sp . , Alternaria alternats et Ulocladium botrytis . Parmi les bactéries, levures et/ou champignons nosocomiaux et/ou qui peuvent causer des maladies, l'on retrouve en particulier certaines souches d' Escherichia coli, de Staphyloccocus aureus, en particulier les souches résistantes à la méticilline (MRSA) , de Pseudomonas aeruginosa et de Candida albicans, voire de Salmonella enterica (subs. Enterica) et Streptococcus pyogenes . Borrelia sp., Corynebacterium sp. , Enterobacter sp. , Escherichia coli, Flavobacterium meninqosepticum, Haemophilus sp. , Helicobacter pylori, Klebsiella sp. , Leqionella sp. , Listeria ivanovii, Listeria monocytogenes, Mycobacterium sp. , Neisseria meninqitidis, Pasteurella sp. , Proteus mirabilis, Pseudomonas aeruquinosa, Salmonella sp (eg S. typhimurium), Shiqella dysenteriae, Staphylococcus aureus, Streptococcus sp. (eg S. pneumoniae or S. pyoqenes) Vibrio sp (eg V. cholerae), Yersinia sp (eg Y. pestis and Y. enterolitica), Asperqillus fumiqatus, Candida albicans, Trichphyton sp, Bacillus cereus, Cladosporidium sp. Penicillium sp. , Asperqillus sp. Alternaria alternates and Ulocladium botrytis. Among the bacteria, yeast and / or nosocomial fungi and / or which can cause diseases, we find in particular certain strains of Escherichia coli, Staphylococcus aureus, in particular meticillin-resistant strains (MRSA), Pseudomonas aeruginosa and Candida albicans or even Salmonella enterica (Enterica subsp.) and Streptococcus pyogenes.
Un autre aspect connexe de la présente invention est l'utilisation (ou l'incorporation) de la bactérie identifiée ci-dessus dans des compositions détergentes. Another related aspect of the present invention is the use (or incorporation) of the bacterium identified above in detergent compositions.
Les compositions détergentes préférées sont celles pour le traitement des surfaces, en particulier des surfaces de collectivités telles que les hôpitaux, les crèches pour enfants en bas-âge, les maisons de repos pour personnes âgées ou en revalidation, les grands magasins (en ce compris les poignées des caddies), les cantines.  The preferred detergent compositions are those for the treatment of surfaces, particularly surfaces of communities such as hospitals, nurseries for young children, nursing homes for elderly or revalidating, department stores (including the handles of caddies), canteens.
Ceci permet avantageusement de nettoyer ces surfaces, tout en réduisant de manière directe l'abondance des pathogènes qui y seraient présents.  This advantageously makes it possible to clean these surfaces, while directly reducing the abundance of pathogens that would be present.
Dans le contexte de la présente invention, les compositions détergentes ne sont pas particulièrement limitées.  In the context of the present invention, the detergent compositions are not particularly limited.
Cependant, les compositions détergentes sont des liquides. Elles ont, de préférence, le pH (en général les compositions détergentes contiennent de l'eau, ce qui n'exclut pas la présence, même majoritaire, de solvants organiques) et les différents composés en concentration qui sont compatibles avec la vie de la bactérie (décrite ci-dessus) pendant une période suffisamment longue (par exemple plusieurs semaines, ou plus ; en général, les compositions sont stables plusieurs mois, par exemple 10 mois ou plus) .  However, the detergent compositions are liquids. They preferably have pH (in general the detergent compositions contain water, which does not exclude the presence, even majority, of organic solvents) and the various compounds in concentration which are compatible with the life of the bacterium (described above) for a sufficiently long period (for example several weeks or more, in general, the compositions are stable for several months, for example 10 months or more).
Si ce n'est pas possible, la bactérie de l'invention peut être formulée dans une composition intermédiaire, de préférence liquide, qui doit être mélangée juste avant usage avec la composition détergente, de préférence dans des proportions simples (ex. entre 1 : 100 et 100 :1, de préférence entre 1 : 10 et 10 : 1 ) . If this is not possible, the bacterium of the invention may be formulated in an intermediate composition, preferably a liquid, which must be mixed just before use with the detergent composition, preferably in simple proportions (eg between 1: 100 and 100: 1, preferably between 1: 10 and 10: 1).
Dans le contexte de la présente invention, la composition détergente est une solution liquide (aqueuse et/ou comprenant un ou plusieurs solvants organiques) et cette solution comprend (outre la bactérie décrite ci-dessus) au moins une molécule ayant des propriété de surface et/ou tensio-active et/ou amphiphile tels que des surfactants et des savons (de préférence moins de 30% en poids) et un (au moins un) agent conservateur (et/ou stabilisant) (jusqu'à 10% en poids pour le total des agents conservateurs présents) . Lorsque la composition détergente est aqueuse, en ce compris une composition comprenant un ou plusieurs solvants organiques, son pH est fixé, de préférence entre 2,5 et 11,5, avantageusement entre 4 et 10, voire entre 7 et 10.  In the context of the present invention, the detergent composition is a liquid solution (aqueous and / or comprising one or more organic solvents) and this solution comprises (besides the bacterium described above) at least one molecule having surface properties and and / or surfactant and / or amphiphilic surfactants and soaps (preferably less than 30% by weight) and one (at least one) preservative (and / or stabilizing) agent (up to 10% by weight for total preservatives present). When the detergent composition is aqueous, including a composition comprising one or more organic solvents, its pH is set, preferably between 2.5 and 11.5, advantageously between 4 and 10, or even between 7 and 10.
La composition comprend en outre, de manière avantageuse, des cires et/ou des polymères (de préférence moins de 30% en poids) , des agents de blanchiment (de préférence moins de 15% en poids) , des acides, bases (préférés) ou sels (de préférence un total de moins de 30% en poids) , ainsi que un ou plusieurs composés choisis dans le groupe constitué des chélatants, enzymes, épaississants et inhibiteurs de corrosion.  The composition further comprises, advantageously, waxes and / or polymers (preferably less than 30% by weight), bleaching agents (preferably less than 15% by weight), acids, bases (preferred) or salts (preferably a total of less than 30% by weight), as well as one or more compounds selected from the group consisting of chelants, enzymes, thickeners and corrosion inhibitors.
La composition détergente peut optionnellement comprendre un ou plusieurs parfum(s) et/ou un ou plusieurs agent (s) de coloration .  The detergent composition may optionally include one or more perfume (s) and / or one or more dyeing agent (s).
Un autre aspect connexe de la présente invention est la bactérie identifiée ci-dessus dans un milieu liquide à une concentration d'au moins 200000 cfu/ml, par exemple au moins 500000 cfu/ml, au moins 1 million cfu/ml, au moins 5 millions cfu/ml, au moins 10 millions cfu/ml, par exemple environ 40 à environ 70 millions cfu/ml. Cfu est l'acronyme en anglais de « colony forming unit », ce qui est la manière usuelle de quantifier la présence d'une bactérie par sa capacité à générer des colonies, une fois cette bactérie ensemencée sur un support de culture approprié. Another related aspect of the present invention is the bacterium identified above in a liquid medium at a concentration of at least 200,000 cfu / ml, for example at least 500,000 cfu / ml, at least 1 million cfu / ml, at least 5 million cfu / ml, at least 10 million cfu / ml, for example about 40 to about 70 million cfu / ml. Cfu is the English acronym for "colony forming unit", which is the usual way of quantify the presence of a bacterium by its capacity to generate colonies, once this bacterium seeded on a suitable culture medium.
De préférence ce milieu liquide comprend en outre au moins une molécule ayant des propriétés de surface et/ou tensio-active et/ou amphiphile tels que des surfactants et des savons (de préférence moins de 30% en poids) et un agent conservateur (stabilisant) (jusqu'à 10% en poids, de préférence jusqu'à 1% en poids) .  Preferably, this liquid medium further comprises at least one molecule having surface and / or surfactant and / or amphiphilic properties such as surfactants and soaps (preferably less than 30% by weight) and a preservative (stabilizing agent). ) (up to 10% by weight, preferably up to 1% by weight).
Le milieu liquide comprend avantageusement en outre un ou plusieurs composés pour favoriser la croissance de la bactérie de l'invention (une fois mise sur une surface à nettoyer), de préférence choisis parmi les nucléotides (ex ; adénine, guanine ; 0,1 à 1% poids : poidsmiiieu liquide)/ une phytohormone , par exemple l'indole 3-acétique (de 0,01% à 0,1 % en poidsmiiieu liquide) l'azote minéral (ex. NH4S04 : 0,1 à 1% en poidsmiiieu liquide) et une source de carbone (ex. le glucose) . Ce composé pour favoriser la croissance de la bactérie de l'invention est particulièrement utile pour les formulations où la concentration de la bactérie de l'invention est plus faible (ex. 200000 cfu/ml, 500000 cfu/ml, 1000000 cfu/ml) The liquid medium advantageously further comprises one or more compounds for promoting the growth of the bacterium of the invention (once placed on a surface to be cleaned), preferably chosen from nucleotides (eg, adenine, guanine, 0.1 to 1% by weight: weight of liquid medium ) / a phytohormone, for example 3-acetic indole (from 0.01% to 0.1% by weight, liquid medium ), mineral nitrogen (eg NH 4 S0 4 : 0, 1-1% by weight liquid miiieu) and a carbon source (eg. glucose). This compound for promoting the growth of the bacterium of the invention is particularly useful for formulations where the concentration of the bacterium of the invention is lower (eg 200,000 cfu / ml, 500,000 cfu / ml, 1000000 cfu / ml)
Le milieu liquide peut être aqueux, ce qui signifie qu'il comprend de l'eau et peut également comprendre des solvants non-aqueux . The liquid medium may be aqueous, which means that it comprises water and may also include non-aqueous solvents.
Un milieu liquide préféré comprend au moins 50% en poids (poids solvant organique :poids de l'ensemble des solvants) d'un solvant (c'est-à-dire une substance non-aqueuse liquide à température ambiante) choisi dans le groupe constitué des alcools (ex. benzylique, éthanol, isopropanol) , des éthers de glycols (ex. butyldiglycol , monopropylène glycol, dipropylène glycol) , des distillats pétroliers (ex. « white spirit », éther de pétrole, huile de paraffine), des composés aminés (ex. monoéthanolamine) , des dérivés cétoniques et aldéhydiques (ex. méthylal, diméthoxy méthane) . A preferred liquid medium comprises at least 50% by weight (organic solvent weight: weight of all solvents) of a solvent (i.e. a non-aqueous liquid at room temperature) selected from the group consisting of alcohols (eg benzyl, ethanol, isopropanol), glycol ethers (eg butyldiglycol, monopropylene glycol, dipropylene glycol), petroleum distillates (eg "white spirit", petroleum ether, paraffin oil), amino compounds (ex. monoethanolamine), ketone and aldehyde derivatives (eg methylal, dimethoxy methane).
Le (s) conservateur ( s ) ( stabilisant ( s ) ) sont présents à des concentrations de 10% en poids maximum, de préférence entre 0,001% et 10%, de manière plus préférée, entre 0,01% et 1%, de manière encore plus préférée, entre 0,05% et 0,2% (poids du ou des conservateur ( s ) présent (s) :poids de la composition).  The preservative (s) (stabilizer (s)) are present at concentrations of 10% by weight maximum, preferably between 0.001% and 10%, more preferably between 0.01% and 1%, of even more preferably, between 0.05% and 0.2% (weight of the preservative (s) present (s): weight of the composition).
Le (s) conservateur ( s ) (stabilisants) sont de préférence choisis parmi le groupe constitué de 1 ' isothiazolinone (ex. 1,2- Benzisothiazol-3-one; 2-Methyl-2H-isothiazol-3-one ; 5-Chloro-2- methyl-isothiazolin-3 (2H) -one (CMI) ; 2-methylisothiazolin- 3 (2H) -one (MI)), du formaldéhyde et de ses libérateurs (ex glutaraldehyde ; 2-bromo-2-nitropropane-l , 3-diol ; 5-bromo-5- nitro-1 , 3-dioxane) , des conservateurs (stabilisants) avec une fonction amine (ex Diazolinidylurea ; Chloroacetamide) , des conservateurs (stabilisants) avec une fonction alcool (ex : Phenoxypropanol ; Benzyl alcohol ; o-Phenylphenol ; Phenoxy- ethanol) , du benzoate et de ses dérivés, des parabènes et dérivés, du methyldibromoglutaronitrile, du sodium hydroxy methyl glycinate, du triclosan et du sorbate (et/ou acide sorbique) , ainsi que des mélanges de plusieurs de ces conservateurs (stabilisants) . Par exemple un mélange CMI + MI dans des proportions massiques de 3:1.  The preservative (s) (stabilizers) are preferably selected from the group consisting of isothiazolinone (eg 1,2-benzisothiazol-3-one; 2-methyl-2H-isothiazol-3-one; Chloro-2-methyl-isothiazolin-3 (2H) -one (CMI) 2-methylisothiazolin-3 (2H) -one (MI)), formaldehyde and its liberators (eg glutaraldehyde, 2-bromo-2-nitropropane -1,3-diol; 5-bromo-5-nitro-1,3-dioxane), preservatives (stabilizers) with an amine function (eg Diazolinidylurea, Chloroacetamide), preservatives (stabilizers) with an alcohol function (ex: Phenoxypropanol, benzyl alcohol, o-phenylphenol, phenoxyethanol), benzoate and its derivatives, parabens and derivatives, methyldibromoglutaronitrile, sodium hydroxy methyl glycinate, triclosan and sorbate (and / or sorbic acid), and mixtures of several of these preservatives (stabilizers). For example a CMI + MI mixture in mass proportions of 3: 1.
Exemples Examples
Exemple 1. Souches pathogènes (SP) .  Example 1. Pathogenic strains (SP).
Exemple 2. Mise au point d'un milieu liquide commun à la bactérie de l'invention et aux SP. L' inventeur a remarqué que toutes les souches SP ci-dessus (même CA, dont la culture est recommandée plutôt à pH 5, 6) , mais également la bactérie de l'invention, sont capables de croitre dans un milieu A comprenant un extrait de levure, du peptone et du glucose à pH 7 et à température ambiante. Example 2. Development of a liquid medium common to the bacterium of the invention and SP. The inventor has noticed that all the SP strains above (same CA, whose culture is recommended rather at pH 5, 6), but also the bacterium of the invention, are capable of growing in medium A comprising an extract yeast, peptone and glucose at pH 7 and at room temperature.
Exemple 3. Analyse de l'interaction entre la Bactérie de l'invention et SP en culture liquide. Example 3. Analysis of the interaction between the bacterium of the invention and SP in liquid culture.
L' inoculum de départ de la souche de la bactérie de l'invention a été fixé à une concentration élevée. Pour ce faire, une colonie de la bactérie de l'invention est prélevée à partir d'une boite agar et inoculée dans 10 ml du milieu A dans une fiole. Le nombre de CFU/ml est déterminé à l'issue d'une incubation de 24h au compteur de particules. The starting inoculum of the strain of the bacterium of the invention was fixed at a high concentration. To do this, a colony of the bacterium of the invention is taken from an agar plate and inoculated into 10 ml of medium A in a vial. The number of CFU / ml is determined after a 24-hour incubation with the particle counter.
La concentration de l' inoculum de départ des souches SP est de 5.102 CFU/ml. Vu la difficulté de mesurer des concentrations aussi faibles au compteur de particules, plusieurs colonies sont prélevées sur boite agar et suspendues dans un volume précis de milieu A. Cette culture est alors dénombrée au compteur de particules et ensuite diluée à la concentration voulue . The concentration of the starting inoculum of the SP strains is 5.10 2 CFU / ml. In view of the difficulty of measuring such low concentrations in the particle counter, several colonies are removed from the agar plate and suspended in a precise volume of medium A. This culture is then counted on the particle counter and then diluted to the desired concentration.
Comme montré aux figures 1-3, en culture liquide, la bactérie de l'invention a réduit la croissance de EC, réduit fortement la croissance de CA et a eu un effet encore plus marqué surAs shown in FIGS. 1-3, in liquid culture, the bacterium of the invention reduced EC growth, greatly reduced AC growth and had an even stronger effect on
MRSA. MRSA.
La bactérie de l'invention a également fortement réduit la croissance en milieu liquide Salmonella enterica (subs. Enterica) et de Streptococcus pyogenes .  The bacterium of the invention also greatly reduced the growth in liquid medium Salmonella enterica (subs Enterica) and Streptococcus pyogenes.
Par contre, la bactérie de la présente invention n'a pas eu d'effet significatif en culture liquide sur la croissance de KP ou de PA, ce qui montre que la bactérie de l'invention n'a pas de propriétés biocides générales, et suggère un effet spécifique . On the other hand, the bacterium of the present invention did not have any significant effect in liquid culture on the growth of KP or PA, which shows that the bacterium of the invention does not have general biocidal properties, and suggests a specific effect.
Exemple 3. Analyse de l'interaction entre la Bactérie de l'invention et SP en milieu solide. Example 3. Analysis of the Interaction Between the Bacterium of the Invention and SP in a Solid Medium
L'interaction entre la bactérie de l'invention et chaque souche de SP a été étudiée en milieu gélosé : d'une part, l'effet du surnageant de culture a été testé sur une culture gélosée de chaque SP ; d'autre part, l'effet de la bactérie de l'invention sur la croissance des souches SP a été testée par étalement en stries ou par ajout de colonies de la bactérie de l'invention sur un tapis de SP. The interaction between the bacterium of the invention and each strain of SP was studied in agar medium: firstly, the effect of the culture supernatant was tested on an agar culture of each SP; on the other hand, the effect of the bacterium of the invention on the growth of SP strains was tested by streaking or by adding colonies of the bacterium of the invention to a SP carpet.
CA et EC ont été mises en culture, et montrent une croissance en tapis. L'ajout du surnageant de culture de la bactérie de l'invention a inhibé la croissance de CA et de EC. Un effet inhibiteur était moins évident dans le cas des autres SP.  CA and EC were cultured, and show carpet growth. The addition of the culture supernatant of the bacterium of the invention inhibited the growth of CA and EC. An inhibitory effect was less evident in the case of other SPs.
La mesure d' interactions par stries a montré une croissance importante de la bactérie de l'invention, et un faible nombre de colonies CA. L'effet est encore plus spectaculaire dans le cas de MRSA, et également observable dans le cas de EC.  Measurement of striated interactions showed significant growth of the bacterium of the invention, and a low number of CA colonies. The effect is even more dramatic in the case of MRSA, and also observable in the case of EC.
L' inventeur a également mis les SP en culture sur agar, puis après séchage, 3 colonies de la bactérie de l'invention. Dans le cas de CA, la bactérie de l'invention a eu une croissance importante, qui recouvre CA. L'inventeur a observé que la bactérie de l'invention, là où elle a été ensemencée, a causé des halos d'inhibition de la croissance de MRSA. L'effet est moins marqué pour les autres SP.  The inventor also cultured the SPs on agar, and after drying, 3 colonies of the bacterium of the invention. In the case of CA, the bacterium of the invention has had a significant growth, which covers CA. The inventor has observed that the bacterium of the invention, where it was seeded, caused MRSA growth inhibition halos. The effect is less pronounced for other SPs.

Claims

Revendications claims
1. Composition détergente liquide comprenant une ou plusieurs molécule (s) ayant des propriétés de surface et/ou tensio- active et/ou amphiphile dans une quantité pondérale comprise entre 1% et 30% ; un ou plusieurs agent (s) conservateur ( s ) dans une quantité pondérale comprise entre 0,001% et 10% et une bactérie antagoniste de un ou de plusieurs micro organisme (s) pathogène (s) ou indésirable ( s ) . A liquid detergent composition comprising one or more molecule (s) having surface and / or surfactant and / or amphiphilic properties in an amount by weight of between 1% and 30%; one or more preservative agent (s) in a weight amount of between 0.001% and 10% and a bacterium antagonist of one or more microorganisms (s) pathogenic (s) or undesirable (s).
2. Composition détergente liquide selon la revendication 1 dans laquelle la bactérie antagoniste de un ou de plusieurs micro organisme (s) pathogène (s) ou indésirable ( s ) agit par compétition avec le ou les micro-organisme ( s ) pathogène (s) ou indésirable ( s ) , ou produit un facteur qui perturbe spécifiquement le métabolisme du ou des micro-organismes pathogène (s) ou indésirable ( s ) .  A liquid detergent composition according to claim 1 wherein the bacterium antagonizing one or more pathogenic or undesirable microorganism (s) acts in competition with the pathogenic microorganism (s) or undesirable (s), or produces a factor that specifically disrupts the metabolism of the pathogenic or unwanted microorganism (s).
3. Composition détergente liquide comprenant une ou plusieurs molécule (s) ayant des propriétés de surface et/ou tensio- active et/ou amphiphile dans une quantité pondérale comprise entre 1% et 30% ; un ou plusieurs agent (s) conservateur ( s ) dans une quantité pondérale comprise entre 0,001% et 10% et une bactérie susceptible d'être obtenue par une méthode comprenant les étapes successives suivantes :  A liquid detergent composition comprising one or more molecule (s) having surface and / or surfactant and / or amphiphilic properties in an amount by weight of between 1% and 30%; one or more preservative agent (s) in a weight content of between 0.001% and 10% and a bacterium that can be obtained by a method comprising the following successive stages:
- on prélève une bactérie à tester ;  a bacterium to be tested is taken;
- on prélève un ou plusieurs micro-organisme ( s ) pathogène (s) ou indésirable ( s ) ;  - one or more pathogenic microorganisms (s) or unwanted (s);
- on détermine les conditions de culture permettant la croissance à la fois dudit ou desdits micro-organismes pathogènes ou indésirables et de ladite bactérie et/ou on prélève un surnageant de culture de ladite bactérie ;  the culture conditions for the growth of both said pathogenic or undesirable microorganism (s) and of said bacterium are determined and / or a culture supernatant of said bacterium is taken;
- on détermine la croissance dudit ou desdits micro organisme (s) pathogène (s) ou indésirable ( s ) (i) en l'absence de ladite bactérie et (ii) en présence de ladite bactérie ou dudit surnageant de culture de ladite bactérie ;  determining the growth of said microorganism (s) pathogenic (s) or undesirable (i) in the absence of said bacterium and (ii) in the presence of said bacterium or said culture supernatant of said bacterium;
- on identifie une bactérie qui provoque une réduction spécifique de la croissance d'au moins un dudit ou desdits micro-organisme ( s ) pathogène (s) ou indésirable ( s ) .  a bacterium is identified which causes a specific reduction in the growth of at least one of said pathogen (s) or unwanted microorganism (s).
4. Composition détergente selon une quelconque des revendications précédentes dans laquelle au moins 100 nucléotides contigus du génome de la bactérie a au moins 75% d'identité avec au moins 100 nucléotides contigus de SEQ.ID.NO :1 ou de SEQ.ID.NO :3 et/ou de SEQ.ID.NO :4 et/ou de SEQ.ID.NO :5. 4. Detergent composition according to any one of preceding claim wherein at least 100 contiguous nucleotides of the genome of the bacterium has at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID.NO: 1 or SEQ.ID.NO: 3 and / or SEQ .ID.NO: 4 and / or SEQ.ID.NO: 5.
5. Composition détergente selon une quelconque des revendications précédentes dans laquelle la bactérie est du groupe Bacillus amyloliquefaciens.  A detergent composition according to any one of the preceding claims wherein the bacterium is from the group Bacillus amyloliquefaciens.
6. Composition détergente selon une quelconque des revendications précédentes, dans laquelle le conservateur est choisi dans le groupe constitué de l' isothiazolinone, du formaldéhyde et ses libérateurs, des conservateurs avec une fonction amine, des conservateurs avec une fonction alcool, du benzoate et de ses dérivés, des parabènes et dérivés, des ammoniums quaternaires, du methyldibromoglutaronitrile, du Sodium hydroxy methyl glycinate, du triclosan et du Sorbate (et/ou acide sorbique) .  A detergent composition according to any one of the preceding claims, wherein the preservative is selected from the group consisting of isothiazolinone, formaldehyde and its liberators, preservatives with an amine function, preservatives with an alcohol function, benzoate and its derivatives, parabens and derivatives, quaternary ammoniums, methyldibromoglutaronitrile, Sodium hydroxy methyl glycinate, triclosan and Sorbate (and / or sorbic acid).
7. Composition détergente selon une quelconque des revendications précédentes, dans laquelle la bactérie est présente à une concentration d'au moins 200000 cfu/ml, de préférence au moins 1000000 cfu/ml, de manière encore plus préférée à une concentration d'au moins 10000000 cfu/ml.  A detergent composition according to any of the preceding claims, wherein the bacterium is present at a concentration of at least 200,000 cfu / ml, preferably at least 1000000 cfu / ml, more preferably at a concentration of at least 10,000,000 cfu / ml.
8. Composition détergente selon une quelconque des revendications précédentes, comprenant en outre un ou plusieurs composé (s) choisis parmi les nucléotides, une phytohormone, l'azote minéral et une source de carbone.  A detergent composition according to any one of the preceding claims, further comprising one or more compounds selected from nucleotides, phytohormone, mineral nitrogen and a carbon source.
9. Composition détergente selon une quelconque des revendications précédentes, dans laquelle le (s) micro organisme (s) pathogène (s) ou indésirable ( s ) comprennent Escherichia coli, Staphyloccocus aureus (MRSA) , et un autre pathogène choisi parmi Pseudomonas aeruginosa, une levure et un champignon, de préférence où un, 2, ou chacun dudit (desdits) pathogène (s) est (sont) résistant (s) à un traitement par antibiotique.  A detergent composition according to any one of the preceding claims, wherein the pathogenic or undesirable microorganism (s) comprises Escherichia coli, Staphylococcus aureus (MRSA), and another pathogen selected from Pseudomonas aeruginosa, yeast and a fungus, preferably wherein one, 2, or each of said pathogen (s) is (are) resistant to antibiotic treatment.
10. Composition détergente selon une quelconque des revendications précédentes, dans laquelle les pathogènes sont connus pour causer des maladies, par exemple des maladies nosocomiales . 10. A detergent composition according to any one of the preceding claims, wherein the pathogens are known to cause diseases, for example nosocomial diseases.
11. Composition liquide comprenant au moins 200000 cfu/ml, de préférence au moins 1000000 cfu/ml, de manière encore plus préférée au moins 10000000 cfu/ml d'une bactérie dont au moins 100 nucléotides contigus de son génome a au moins 75% d'identité avec au moins 100 nucléotides contigus de SEQ.ID. NO :1 ou de SEQ.ID.NO :3 et/ou de SEQ.ID.NO :4 et/ou de SEQ.ID.NO :5. 11. A liquid composition comprising at least 200,000 cfu / ml, preferably at least 1000000 cfu / ml, more preferably at least 10,000,000 cfu / ml of a bacterium of which at least 100 contiguous nucleotides of its genome has at least 75% identity with at least 100 contiguous nucleotides of SEQ.ID. NO: 1 or SEQ.ID.NO: 3 and / or SEQ.ID.NO: 4 and / or SEQ.ID.NO: 5.
12. Composition liquide selon la revendication 11 comprenant en outre un agent conservateur.  The liquid composition of claim 11 further comprising a preservative.
13. Composition liquide selon la revendication 11 ou 12 comprenant en outre un ou plusieurs composé (s) choisis parmi les nucléotides, une phytohormone, l'azote minéral et une source de carbone.  The liquid composition of claim 11 or 12 further comprising one or more compounds selected from nucleotides, phytohormone, mineral nitrogen and a carbon source.
14. Utilisation de la composition détergente selon une quelconque des revendications précédentes 1 à 10 ou de la composition liquide selon une quelconque des revendications précédentes 11 à 13 pour le nettoyage de surfaces, de préférence à l'usage des collectivités, de préférence choisies parmi les hôpitaux, les crèches pour enfants en bas- âge, les maisons de repos pour personnes âgées ou en revalidation, les grands magasins, les cantines et les cuisines .  14. Use of the detergent composition according to any one of the preceding claims 1 to 10 or the liquid composition according to any one of the preceding claims 11 to 13 for cleaning surfaces, preferably for use by communities, preferably chosen from the hospitals, nurseries for infants, retirement homes for the elderly or in revalidation, department stores, canteens and kitchens.
15. Méthode de nettoyage de surfaces de préférence présentes dans les collectivités, de préférence choisies parmi les hôpitaux, les crèches pour enfants en bas-âge, les maisons de repos pour personnes âgées ou en revalidation, les grands magasins, les cantines et les cuisines, comprenant l'application de la composition détergente selon une quelconque des revendications précédentes 1 à 10 ou de la composition liquide selon une quelconque des revendications précédentes 11 à 13 sur lesdites surfaces.  15. Method of cleaning surfaces preferably present in communities, preferably selected from hospitals, nurseries for young children, retirement homes for the elderly or in revalidation, department stores, canteens and kitchens , comprising applying the detergent composition of any of the preceding claims 1 to 10 or the liquid composition of any of the preceding claims 11 to 13 to said surfaces.
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