Composition for releasing and sampling microorganisms

Abstract

Sterile sampling composition comprising a surfactant, one or more enzymes selected from the group consisting of a protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase, a sampling kit comprising this composition, a sampling method based on this composition or kit, and use of this composition or kit for the detection of bacteria on a surface.

Description

[0001]COMPOSITION FOR REMOVING AND COLLECTING MICROORGANISMS Technical field The present invention relates to a composition for removing and collecting microorganisms, to its use, to a kit for removing, collecting and preserving microorganisms comprising said composition and to a method for removing, collecting and preserving microorganisms. Prior art In many fields of activity such as the agri-food sectors, communities, medical and veterinary sectors, problematic contaminations due to the presence of microorganisms are observed very frequently. Conventionally, in hospitals and in medical or veterinary practices, the presence of microorganisms is responsible for nosocomial diseases while in the agri-food and community sectors,These microorganisms are responsible for the degradation of perishable goods but also for the transfer of contaminants to consumers of products from, for example, a meat, fruit, vegetable or other production chain. However, nowadays, strict hygiene standards are imposed and it is therefore necessary to ensure that the number of microorganisms responsible for such contamination is kept below an acceptable threshold value, this acceptable threshold value being specific to each field of activity. Planktonic bacteria are a prime example of contaminating microorganisms, these bacteria are free at the level of a liquid or solid substrate and pose a problem in themselves because they are capable of directly contaminating any type of surface such as foodstuffs, medical tools, conveyor belts, storage tanks or even human patients or animals themselves. However, today,It is widely recognized that the problems related to contamination by microorganisms are all the more important as these form biofilms. Indeed, in all types of industry and more particularly in the food industry, biofilms are inevitably formed, given the richness of the surrounding environment. Biofilms are defined as a consortium of microorganisms embedded in a matrix of extracellular polymeric substances. In other words, biofilms are viscous films that develop on all surfaces, following the adhesion of microorganisms on these surfaces and the secretion by them of polymers covering them,facilitating their adhesion to the surface and thus forming the extracellular matrix. Biofilms thus constitute a protective layer around microorganisms that is highly resistant to chemical and thermal aggression, which makes their elimination very difficult using conventional biocides and also represents a recurring and significant source of contamination of the surrounding environment. This resistance is explained on the one hand by the matrix of extracellular polymeric substances that forms a physical barrier to the diffusion of biocidal molecules in the biofilm and, on the other hand, by the sessile state of the microorganisms and their ability to exchange genes responsible for certain biocide resistance mechanisms very quickly. Therefore, it is easy to understand that biofilms are very resistant and extremely varied structures; for example, different strains of the same bacterium can form different biofilms,These biofilms are even more varied as they are composed of several different bacteria. In addition, bacteria exchange resistance genes among themselves and the environment also influences the biofilm, further accentuating their complexity and resistance. In addition, the extracellular matrix of biofilms can be identified when it is highly developed but, in the majority of cases, the biofilm develops insidiously in the facilities and its presence (or the presence of problematic microorganisms) will only be detected during the analysis of the quality of the finished product. Regarding the growth of the biofilm, this takes place in a cyclical manner comprising a growth phase during which the accumulation of microorganisms occurs and a detachment phase during which whole pieces of biofilms and microorganisms detach by erosion and under the effect of their weight to contaminate surfaces, foodstuffs,medical tools, conveyor belts, storage tanks or even human patients, consumers or the animals themselves. This must lead to the industrialist stopping their production line, carrying out a cleaning cycle to eliminate the biofilm, representing many hours of work, deployed resources and loss of yield. From all this, it emerges that contaminating microorganisms and / or biofilms constitute a real problem, particularly in the fields of healthcare (hospitals, dental or medical practices), veterinary care and the food industry. This problem is all the more critical since microorganisms and / or biofilms can involve bacteria responsible for potentially fatal infections in individuals, whether these bacteria are present in hospitals,veterinary practices or even in the food industry and ultimately end up on food products intended for consumption. As indicated above, the problem of microorganisms and more particularly biofilms is twofold. On the one hand, conventional disinfectants and biocides are very often ineffective because they fail to reach the microorganisms protected by the extracellular matrix of the biofilm, this matrix being of complex and highly variable structure and composition. On the other hand, a biofilm is generally mixed, that is to say it contains a multitude of different bacteria or the same bacteria but which are of different strains, which promotes the spread of resistance genes between the bacteria in the biofilm and thus makes their treatment very difficult or even ineffective. Consequently, from one environment to another or from one sector of activity to another,It is quite common for the detected biofilms to be completely different. In order to detect biofilms, there are kits for detecting the presence of biofilms on surfaces or in more specific installations (water circuits, etc.), these kits (such as the one disclosed in document EP2537601) allow selective staining of biofilms. It is therefore currently possible to determine areas where biofilms are present in order to eliminate them without knowing the precise nature of the microorganisms (bacteria) at the origin of the targeted biofilm. As a result, biofilm removal compositions comprising several enzymes are used without knowing whether the enzymes used and possibly formulated in a detergent base (see for example document EP2243821) are actually adequate to act on a given biofilm. For this reason,Current treatments are rather random and non-specific, which is economically unprofitable and can lead to a considerable loss of time and downtime of installations. Furthermore, techniques for sampling microorganisms from surfaces have been developed in order to characterize and / or quantify the populations of microorganisms present on surfaces and responsible for contamination. These sampling methods therefore make it possible to determine, on a surface, the different types (of strains) of bacteria and microorganisms present. Among the methods for sampling microorganisms present on a surface,One of the references in this area is the ISO 18593:2004(F) Standard, which provides and defines horizontal methods for sampling techniques on surfaces (more specifically on surfaces found in the environment of the food industry). The "cloth / sponge" method is an example of sampling techniques cited in this Standard, this method making it possible to search for and / or count microorganisms present on a surface. Briefly, according to this sampling method, it involves wetting the cloth / sponge with a quantity of diluent which is sterile physiological serum, sampling the surface in two perpendicular directions before introducing the cloth / sponge into a sterile container with the diluent. Subsequently, after possible storage of the sample, the latter is analyzed quantitatively and / or qualitatively. Unfortunately,if such a method allows for the collection of free microorganisms on the surface of a substrate, i.e. a collection of planktonic microorganisms, it turns out to be ineffective when the surface is contaminated by microorganisms that have formed a biofilm. Indeed, as explained above, the microorganisms protected by the extracellular matrix of the biofilm, this matrix being of complex and highly variable structure and composition and biofilms being mixed, they are even more varied and complex as they are composed of several different bacteria or different strains. In addition, it turns out that sampling techniques based on a support impregnated with water can have a beneficial effect on the development of microorganisms, even when sampling from a surface. Indeed, it is recognized that certain bacteria exhibit increased growth in the presence of water,the water-soaked support therefore promoting this growth during sampling but also after it since a thin film of water remains on the treated support where the sampling took place. This can therefore promote the development of certain non-sampled microorganisms, in particular when they are protected by a biofilm. In order to overcome the disadvantages of water-soaked sampling supports, document DE10304331 is known from the state of the art, which discloses an enzymatic preparation that can be used to eliminate biofilms from surfaces in the absence of biocides. This enzymatic preparation comprises one or more enzymes from the group of polysaccharidases and / or proteases and possibly nucleases. Unfortunately, even if the compositions according to document DE10304331 have a certain effectiveness in terms of sampling microorganisms,it appears that they are insufficient given the diversity and complexity of the microorganisms and biofilms that may be present on a surface of an installation and that they do not allow a representative sampling of the bacterial populations that are nevertheless present there, that is to say that they only allow a low coverage of the microorganisms. Indeed, the biofilms and / or microorganisms present on a surface of an installation are on the one hand composed of a varied and complex population of microorganisms, that is to say different microorganisms but also these different microorganisms can still be of different strains, this leading to different resistances but also the formation of varied, complex and different extracellular matrices of biofilms,sometimes within the same surface of an installation. There is therefore a real need to provide a composition which aims to overcome the disadvantages of the state of the art by allowing efficient, representative and adequate sampling of microorganism populations, whether free but also in the form of a biofilm with a complex structure which can vary considerably from one sample to another. In addition, there is a need to provide a composition allowing the sampling of microorganisms on different types of surfaces, in different environments, stable over time and which is easy to use by users or even compatible with the production chain, for example food in an agri-food industry. In this respect, patent application WO 2018 / 002013 describes a viscous aqueous solution comprising one or more enzymes and 8,4% surfactants for application to a horizontal or even vertical surface, for prolonged impregnation of a fabric placed on this surface, so as to recover a maximum of any microorganisms that may be present, including in the form of biofilm, while preserving their viability and quantifying their presence. In another technical field, patent EP0578666 discloses a composition for dishwashers comprising surfactants, enzymes and preservatives. The composition may, in one embodiment, be in liquid and concentrated form. However, this publication does not describe polyol-type stabilizing agents and provides information on the use of substances that are not compatible with sampling. Similarly, patent US5571504 discloses a composition for the treatment of contact lenses comprising a significant amount (1.75%) of detergents. Furthermore, again,the presence of polyol-type stabilizing agents is not disclosed. Patent application US2019 / 256803 describes a composition for the destruction of biofilms. This document does not describe the use of polyol-type stabilizers, or even sequestering agents. Finally, patent JP5466782 describes a cleaning composition having anti-biofilm efficacy and comprising a protease, propylene glycol and surfactants. However, this document does not describe a sampling composition: the surfactant contents, or even their compatibility with the sampling, are, for example, not specified. Brief summary of the invention The present invention relates to a (sterile) composition for sampling microorganisms present on a surface, the sampling being compatible with downstream microbiological tests, comprising: one or more surfactants, present at a content of at least 0.01% by weight and at most 1,5% by weight relative to the weight of said composition (sum of the weights of surfactants: weight of the composition), this or these surfactants comprising (or consisting essentially of) an anionic surfactant; at least two enzymes chosen from the group consisting of a (serine) protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase; at least one stabilizing agent present at a content of at least 15% relative to the weight of said composition, said stabilizer being chosen from the group of polyols (preferably glycerol) and, preferably at least one sequestering agent. The present invention also relates to the use of this composition for sampling for the detection of microorganisms, advantageously Salmonella sp. and / or Listeria sp. Detailed description of an embodiment of the invention The techniques for evaluating the presence of microorganisms include culture,biochemistry (measuring ATP or NADH production, measuring enzymatic activity, for example catalase, or even measuring proteins in a medium), genetic methods of "Polymerase Chain Reaction" (PCR) and sequencing (e.g. 16S ribosomal RNA) and immunological methods. In other words, many enzymes are used for biochemical analysis or for molecular biology analyses, flow cytometry or immunoassays, such as alkaline phosphatase, Taq polymerase, luciferase, etc. However, inventors noticed that residual proteolytic activity interfered with these tests. In addition, proteases interfered with antigen-antibody complexes during immunological type reactions, also widely used for the detection of microorganisms. Often several methods are combined,in particular the detection of pathogenic microorganisms potentially present at low intensity comprises a culture step, followed by immunological and / or genetic marking. This implies that the sampling of the microorganisms must ensure sufficient recovery, sufficient viability, but also that the products used do not interfere with the tests to be carried out downstream. There is therefore a very delicate balance to be respected. To solve this problem, the invention provides a sterile composition for sampling, detaching and / or collecting microorganisms potentially present on a surface comprising: - at least one anionic surfactant, the surfactant(s) together representing at least 0.01% (preferably at least 0.02%) by weight relative to the weight of said composition (weight of surfactants: weight of the solution), and at most 1.5%, preferably at most 1%, or even 0.5%, or even 0,1%; - at least one polyol (glycerol) as stabilizing agent and / or viscosifying agent representing at least 15%, preferably at least 18%, preferentially approximately 20% by weight relative to the weight of said composition, and / or preferably less than 30%, or even less than 25%, by weight relative to the weight of said composition, - advantageously, at least one sequestering agent (di- or multi-valent cations), preferably in a weight content of at least 0.2%, preferably at least 0.3%, or even at least 0.4%; - at least two enzymes (in a concentration capable of degrading any bacterial biofilms or soiling and in a concentration which does not interfere with downstream microbiological tests), said at least two enzymes being chosen from the group consisting of a (serine) protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase. - Preferably,the surfactant(s) present in the collection composition (e.g. at a content of between 0.02% and 1.5%, or maximum 1%, maximum 0.5%, maximum 0.1%) consist (essentially) of one or more anionic surfactants. This composition preferably has (i) a pH of between 5 and 11, preferably between 6 and 10, even more preferably between 7 and 9, or between 7.5 and 8 and / or (ii) a dynamic viscosity (20°C) of between 1, preferably 2, 5, 10, 20, 30, 40, 50 mPa.s and 150, preferably 100, 90, 80, or even 70 mPa.s. In the context of the present invention, the word "surface" is understood broadly and preferably means a surface of objects or premises of individuals, industry, or even public spaces (including public transport or public toilets) potentially contaminated by microorganisms,including pathogenic bacteria or microorganisms. For example, many surfaces in the food industry risk being contaminated by potentially harmful microorganisms (toxic or spoiling), which is intolerable and forces, if necessary, the removal of infected batches. Among these surfaces, we can find tables, utensils, handles. Pharmacies, pharmaceutical industries, hospitals, medical practices also have many surfaces potentially contaminated by potentially harmful microorganisms, which must be identified quickly and precisely, which the present invention allows. Among the microorganisms whose rapid and / or precise identification is of interest, we find, for the food industry, pathogenic microorganisms (for example, Escherichia Coli, Salmonella sp., Listeria monocytogenes, Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa,Campylobacter jejuni, Clostridium perfringens, Aspergillus flavus), spoilage microorganisms (e.g., Brochotrix thermosphacta, Lactaobcillus rahmnosus, Lactococcus piscium, Leuconostoc carnosum / gelidum, Paenibacillus sp, Zygosaccharomyces bailii / rouxi, Candida sp), or ferments (Lactobacillus casei, Lactococcus lactis, Lactobacillus sakei, Streptococcus thermophilus, Pédiococcus acidilactici, Staphylococcus xylosus, Saccharomyces cerevisiae). More generally, microorganisms potentially present on (non-food) surfaces and of interest to identify quickly and / or precisely include, for example, Escherichia coli, Salmonella sp., Pseudomons sp (e.g. P. aeruginosa), Staphylococcus sp. (e.g. S. aureus), Candida albicans, Alicyclobacillus sp., Stenotrophomonas sp., Acinetobacter sp., Klebsiella sp., Serratia sp., Bacillus sp., Ralstonia picketi and Burkholderia cepacia. Unlike conventional cleaning or sampling solutions,The detergents of the present invention are advantageously chosen so as not to affect the viability of the bacteria to be sampled: both their content and their chemical structure are important. Thus, their concentration is less than 1.5% (weight of all the detergents: weight of the entire composition). However, a minimum content, for example at least 0.01 or at least 0.02 or even 0.03% is necessary. Among the preferred detergents are laureth sulfate (or other "mild" anionic detergents, ideally a hydrocarbon chain of 12 to 14 carbon atoms, a poly(oxyethane-1,2-diyl) segment and an anionic head, for example a sodium sulfate). Those skilled in the art know other anionic detergents that are "mild", or are even able to test other anionic detergents that are considered "mild", for example by means of an in vitro test on microorganism culture (see below). Preferably,the HLB index of all the emulsifiers in the composition is between 10 and 16. In the context of the present invention, the terminology HLB index (for "Hydrophilic-lipophilic balance") relates to the hydrophilic or lipophilic character of the added surfactants. Advantageously, the composition has been sterilized by physical means, for example by irradiation, such as gamma rays. An advantageous alternative consists of sterilizing the composition by sterilizing filtration (e.g. passage through a filter whose meshes have a maximum diameter of 0.22 µm) and / or by UVc irradiation (approximately 200 to 240 nm). Thus, sterilizing filtration of the composition, followed by its incorporation into a bottle (or other container), followed by irradiation of this bottle (the container and the contents) with UVc is preferred because it ensures good sterilization of the contents, and also of the container, which is valuable,for example for use in sensitive areas (hospitals, certain food industries, certain laboratories, etc.). In addition, the inventors have noted that the stability of the enzymatic composition is little affected by this type of irradiation. An alternative to sterilizing filtration (or in synergy with it) is formulation from quasi-sterile components. For example, formulation from water containing less than 10, 6 , preferably less than 10 5 , preferably less than 10 4 , preferably less than 10 3 , preferably less than 10 2, preferably less than 10 CFU (colony forming units) per liter is preferred. This formulation made from this quasi-sterile water, or the bottles comprising it, can then advantageously be subjected to sterilizing irradiation, as described above (gamma, low intensity, electron beam, UVc). Preferably, this composition does not contain a preservative. Alternatively, any preservative that may nevertheless be present must be as diluted as possible, or neutralized quickly (before application of the composition to a surface or just after), so as not to interfere with the metabolism of the microorganisms sampled. Thus, the inventors have noted that compositions, where the usual preservatives, including phenoxyethanol, were present at less than 50 ppm, were acceptable.Preferred compositions advantageously have a lower preservative concentration, for example less than 20 ppm, less than 10 ppm, less than 5 ppm (part per million: weight of the sum of the preservatives (e.g. phenoxyethanol): total weight of the composition), or even a concentration which is below the detection limit and / or 0.1 ppm. Thus, the choice of the constituents (e.g. enzymes) forming the composition is based in part on their preservative content, the lowest possible and / or non-interfering preservatives. Thus, a low residual content of 2-phenoxyethanol is acceptable, although it is best to keep it as low as possible. This sampling composition is preferably for application to a surface, such as a surface used in the food industries.The composition can advantageously be applied to other industrial surfaces such as, for example, the pharmaceutical industry, the healthcare environment or even petrochemicals. The characteristics of the composition according to the present invention have the advantage of providing a composition for sampling microorganisms, whether they are in planktonic form or in the form of complex and varied biofilms. This composition allows for precise sampling of microorganisms. The composition according to the present invention, by comprising at least two enzymes chosen from the group comprising a protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase, advantageously makes it possible to effectively degrade and deconstruct the extracellular matrix of biofilms and in particular of a varied and complex population of biofilms having different extracellular matrices, thus making it possible to release the bacteria in order to sample them.However, the inventors noted that the use of enzymes was particularly delicate when the microorganisms to be sampled were sensitive and in small quantities. Thus, the viability of certain bacteria, particularly Gram-positive ones, is affected by many compounds: downstream tests that involve the culture or metabolism of such a bacterium would give false negative results, which is unacceptable (a "false positive" is less serious than a "false negative," the latter of which can be fatal). Indeed, Listeria is a dreaded contaminant, and the inventors noted that many of the components used prevented its detection downstream, which is all the more pernicious since this is accompanied by a good recovery of other microorganisms, which suggests that the process would have worked well.Beyond Listeria, the inventors have noticed that other Gram-positive bacteria are very sensitive to the various components of the sampling solutions commonly used. Preferably, the composition comprises at least one protease, such as a serine protease (e.g., a subtilisin), the at least one other enzyme being selected from a polysaccharidase, a laccase, a lipase, a cellulase, and a mannanase. The use of a protease greatly increases the sampling capacity of the composition, but, as stated above, the inventors have noticed that this enzyme interferes with downstream tests and even, in some cases, with the other components of the sampling composition itself. Certainly, protease activity can easily be inhibited, at least under laboratory conditions.However, the inventors have noticed that, in practice (on-site, out of the laboratory), the available protease inhibitors are either complicated to use, ineffective, or themselves interfere with downstream microbiological tests. Thus, after numerous unsuccessful tests, the inventors favor the incorporation of a low-intensity protease activity. In the case of Alcalase (e.g., Alcalase 2.5 DX), this is preferably about 0.5 to 5 Anson Units per kg of the composition (0.2 g to 2 g of Alcalase 2.5 DX / Kg of composition), for example, about 1 Anson Unit per kg of the composition. Other proteases, including other commercial batches of Alcalase, may be used, for example, serine proteases.The activity to be incorporated either corresponds to that mentioned above for alcalase (2.5 DX), or is determined in light of the examples of the present invention: the highest concentration that does not interfere with downstream microbiological tests. The person skilled in the art is well aware of how to test for proteolytic activity, for example by using a substrate whose coloration (or fluorescence properties) change after proteolytic cleavage. Thus, determining an activity equivalent to that of a certain amount of alcalase 2.5 DX is within his reach. Alternatively, or in addition, preferably, the substrate used is N-succinyl-alanyl-alanyl-propyl-phenylalanyl-p-nitroanilide (CAS 70967-97-4), which is dissolved in a phosphate buffer at pH 7, 25 mM at a concentration of 1 mM.1 ml of this substrate brought to 20°C is placed in the presence of 100 µl of the solution to be tested (also at 20°C), for example at pH 7, or at an optimal pH for enzymatic activity and the absorbance is measured as a function of time. The enzymatic activity is then easily determined downstream, for example by the Lambert-Beer law. In other words, in light of the above, determining an equivalent activity represents a routine activity. For example, an activity equivalent to 0.2; 0.3; 0.4; 0.5; 0.6; 0.7; 0.8; 0.9; 1; 1.5 or 2 g / Kg of alcalase 2.5 DX advantageously means, in the context of the present invention, that the protease considered (e.g. a serine protease, or even another subtilisin) has the same activity under the conditions described above (CAS substrate 70967-97-4).In light of the present invention, a routine test can be developed, either as an alternative or in combination, by mixing several dilutions of a Listeria culture with the above low amount of alcalase, or with a saline solution or with another protease considered (at several concentrations), followed by measurement by a specific Listeria detection test: the saline composition and the alcalase (applied at a low dose) representing two positive controls. In the non-preferred case where a protease inhibitor is added after sampling, the protease concentrations can be revised upwards. However, the nature and concentration of the protease inhibitor must, particularly preferably, be determined so as not to interfere with downstream microbiological tests.Furthermore, the sequestering agent as well as the pH between 5 and 11 (between 7 and 8.5) of the composition according to the present invention advantageously makes it possible to form complexes with mineral ions which will be fixed in a form preventing their precipitation by the usual reactions and thus to stabilize over time the composition according to the present invention, in particular in view of the enzymes present in such proportions. In the context of the present invention, the sequestering agent is preferably a chemical substance having the capacity to form complexes with mineral ions which it fixes in a form preventing their precipitation by the usual reactions.For example, the sequestering agent may be glucono-delta-lactone, sodium gluconate, potassium gluconate, calcium gluconate, citric acid, phosphoric acid, tartaric acid, sodium acetate, sorbitol, a compound containing a phosphorus atom, or even carboxymethyl inulin (the sodium salt). So-called "mild" sequestering agents are preferred: the inventors have noted that these "mild" sequestering agents allow for better enzymatic activity at the sampling level and / or for downstream microbiological tests involving enzymes (e.g., the inventors have noted interference with downstream PCR tests when too much sequestering agent and / or too strong sequestering agents are present).Alternatively, the sequestering agent may be a phosphorus oxide such as a phosphonate, a phosphinate or a phosphate or a mixture thereof, or a salt thereof, an amine or an amine oxide carrying at least, in its structure, one phosphine, phosphine oxide, phophinite, phosphonite, phosphite, phosphonate, phosphinate or phosphate functional group, alone or in combination, or a salt thereof. Carboxymethyl inulin is advantageous and the inventors have shown that this compound does not interfere with downstream microbiological tests. Similarly, mixtures of carboxymethyl inulin with a glycolate (e.g. in a mass ratio of between 0.5:1 and 2:1; carboxymethyl inulin: sodium glycolate) are preferred. The inventors have also obtained good results with phosphonates.In the context of the present invention, it has been determined that the composition according to the invention, comprising at least one anionic surfactant (comprising one or more surfactants, said surfactant(s) being (essentially) one or more anionic surfactants) which represents at least 0.01% by weight relative to the weight of this composition (preferably at least 0.02%, and less than 1.5%, preferably less than 1%, or even less than 0.5%, or even less than 0.1%), makes it possible in a particularly advantageous manner to detach and collect a representative and complete proportion of a population of microorganisms present on a surface, whether they are planktonic or in the form of biofilms since the enzymes of the composition according to the invention allow their release. It is therefore a delicate balance between the nature of the detergents, their low concentration, the nature of the enzymes and a low activity (protease).Advantageously, this anionic surfactant consists of a foaming surfactant or a foaming surfactant is incorporated into the anionic surfactant(s), this having the advantage on the one hand of detaching and collecting the microorganisms and on the other hand of trapping the microorganisms collected in the composition according to the invention in order to carry out an effective and representative sampling of the population of contaminants of a surface. In addition, the composition according to the present invention comprising at least one anionic surfactant and at least one polyol or propylene glycol as a stabilizing agent and / or viscosifying agent at at least 15% by weight, is stable over time.Furthermore, it has been determined that the composition according to the present invention, by not being too viscous, makes it possible to significantly reduce the contact time required with the microorganisms and / or biofilms present on a surface so as to destructure them sufficiently to release the bacteria and thus be able to sample them and / or identify them for the specific and appropriate subsequent treatment of the surfaces. In other words, the composition according to the present invention therefore makes it possible to completely surround the microorganisms or biofilms and to destructure them more quickly and more efficiently, thus providing an adequate and optimal sampling composition, and also making it possible to trap the microorganisms and contaminants sampled from a surface without the problem of rapid evaporation of the liquid sampling compositions being encountered.As a result, the composition according to the present invention, unlike current compositions, makes it possible to obtain a truly representative and sufficiently complete sample of the contaminants present on a surface, for different types of surfaces and for different environments. This is particularly advantageous because by having a representative and complete sample of the microorganisms, these can subsequently be identified and a targeted and effective elimination treatment, using the appropriate compounds and no longer a cocktail of compounds which is often ineffective, can be applied to the surface. This of course represents a considerable economic and time advantage.The interest of the composition according to the present invention is therefore twofold in that it makes it possible, on the one hand, to collect and sample all the contaminants from a surface, whether planktonic or in the form of biofilms, for analysis and then makes it possible to determine precisely which treatment for removing these contaminants will be applied. Preferably, as described above, the at least one anionic surfactant of the composition according to the invention is chosen from the group comprising sodium laureth sulfate. This molecule has the CAS number 9004-82-4. It is preferably sodium laureth-2 sulfate and / or sodium laureth-3 sulfate. Preferably, the at least one polyol or fatty alcohol as stabilizing agent and / or viscosifying agent is glycerol.The inventors have noticed that glycerol (or other polyols), when present in large quantities (for example more than 15% by weight), increases the stability of the composition over time. Preferably, the composition according to the invention further comprises a pH regulating agent. Preferably, the composition according to the invention has a pH of between 6 and 10, preferably between 7 and 9, preferentially between 8 and 9, particularly advantageously between 8.2 and 8.6. Preferably, the composition according to the invention is in liquid form, preferably in sprayable liquid form. Preferably, the composition according to the invention further comprises at least one additional enzyme chosen from the group consisting of oxidoreductases, lyases, transferases, lipases, esterases, glucosaminidases.As explained above, the interest of the composition according to the present invention is twofold in that it allows on the one hand to collect and sample all the contaminants from a surface whether they are planktonic or in the form of biofilms and that it then allows to precisely determine the nature of these contaminants, which makes it possible to know which treatment for eliminating these contaminants will be applied. Consequently, it is important that the composition according to the invention, which contains the contaminants, microorganisms and biofilms following sampling, can allow a reliable, correct and representative analysis of the sampled population without said composition after sampling interfering with subsequent analyses. The invention therefore also relates to a kit for sampling, detaching, collecting and preserving microorganisms (in a living or revivable state) for the purpose of subsequent laboratory analyses.This kit includes the composition described above, and a means of controlled application thereof. For example, a sprayer calibrated to spray the same quantity of composition, with the same dispersion and in droplets of identical volume.In other words, the present invention provides a kit for sampling, removing (from a surface), taking samples and preserving microorganisms in a living or revivable state for subsequent laboratory analyses comprising: - a composition for sampling (removing and taking samples) microorganisms according to the present invention, - at least one device for controlled application of this composition for sampling (removing and taking samples) on the surface; - preferably at least one means for sterile harvesting of these microorganisms, - preferably at least one sterile container, for example a sterile sachet, comprising a solution for preserving microorganisms for subsequent laboratory analyses, - preferably, this kit further comprises means for specific detection and / or specific quantification by immunology or genetics of bacteria, preferably chosen from Salmonella (e.g.Salmonella enterica, Salmonella bongori), Pseudomonas (e.g. Pseudomonas aeruginosa) and / or Listeria (e.g. Listeria monocytogenes). Preferably, the at least one means for sterile harvesting of the microorganisms of the kit according to the present invention is a tissue, a wipe or a sponge. The preservation solution, in the context of the present invention, preferably comprises polyoxyethylene sorbitan and / or a phosphoglyceride derivative (e.g. lecithin); this composition preferably has the function of neutralizing any potential biocidal activity that would be present in the sampling composition, for example caused by the presence of preservatives that would always be present or disinfecting agents accidentally present. Preferably, the preservation composition comprises a phosphate buffer (e.g. pH 7, 25 mM) which may advantageously further comprise salts, such as magnesium chloride and / or calcium chloride.This preservation solution may advantageously further comprise a protein hydrolysate (e.g. peptone) or a protein-rich compound, so as to provide nutrients to the sampled microorganisms and / or to counteract the deleterious effects associated with residual protease activity, if present. This preservation solution advantageously comprises reducing thiol groups, for example via cysteines (in reduced form) and / or glutathione. Thiols which are not of this nature are also possible, provided that they remain compatible with the metabolism of the microorganisms. When the thiol group is cysteine, it may be present via the protein hydrolysate, for example a protein hydrolysate rich in cysteine ​​and / or where the reducing potential has been ensured. Preferably the polyoxyethylene sorbitan derivative of this kit is polyoxyethylene sorbitan monooleate and / or the phosphoglyceride derivative is phosphatidylcholine.Advantageously, the polyoxyethylene sorbitan derivative, when present, is present at a content of at least 4 g / L, preferably at a content of at least 4.3 g / L, preferentially at a content of at least 4.6 g / L, advantageously at a content of at least 4.9 g / L. Similarly, the phosphoglyceride derivative, when present, is present at a content of between 0.5 and 3 g / L, preferably at a content of between 0.5 and 2.5 g / L, preferentially between 0.5 and 2 g / L, advantageously between 0.5 and 1.5 g / L. Preferably, the protein hydrolyzate (peptone) is present at a content of at least 10 g / L, preferably at least 11 g / L, preferably at least 12 g / L, preferentially at least 13 g / L, advantageously at least 14 g / L, particularly advantageously at least 15 g / L.The invention further relates to a method for sampling, removing, collecting and preserving microorganisms in a living or revivable state for subsequent laboratory analyses comprising the following steps: a) at least one controlled application of a composition for sampling, removing and collecting microorganisms according to the present invention, b) allowing this sampling (removing) composition to act on this surface to be sampled, c) depositing at least one means for collecting these microorganisms on this surface to be sampled, d) allowing this collecting means to be soaked with this sampling (removing) composition, e) sterilely collecting this soaked microorganism collecting means and depositing it in at least one sterile container, for example a sterile bag comprising a solution for preserving microorganisms in a living or revivable state for subsequent laboratory analyses.Advantageously, step b) of the method according to the invention takes place for a duration of at least 10 seconds, at least 20 seconds, at least 30 seconds, at least 1 minute, preferably at least 2 minutes, preferably at least 3 minutes, advantageously at least 4 minutes, particularly advantageously at least 5 minutes and preferably less than 30 minutes, preferably less than 15 or 10 minutes. Preferably, the method according to the invention further comprises at least one preliminary step of preheating this sampling (detachment) composition, preferably to a temperature of between 20 and 60 °C, preferably between 30 and 50 °C, advantageously between 40 and 50 °C. Preferably, the method according to the invention further comprises at least one step of storing this sterile container containing this soaked harvesting means, at a temperature below 15°C, preferably below 10°C, advantageously below 5°C, such as 4 to 8°C.Alternatively, storage may be carried out at less than 0°C, provided that microbiological activity can be restored later. Preferably the controlled application of a volume of the sampling composition is an application of a content of between 0.5 and 2 grams of composition (per 100 cm²), preferably of a content of between 0.7 and 1.8 grams of composition per application, preferably between 0.9 and 1.6 grams of composition (per 100 cm²), advantageously between 1.1 and 1.4 grams of composition (per 100 cm²). Preferably, the sterile collection means is a tissue, a wipe, a swap, a scraper, a swab or a sponge.Advantageously, the subsequent laboratory analyses of step e) described above comprise one or more of the following tests: a test involving a microorganism culture step; a biochemical test (e.g. ATP-metry, NADH detection, catalase activity measurement); a test comprising an immunological recognition step, and a molecular biology test (a test step involving markers and / or probes and / or genetic primers and a combination of these steps, sequencing). As will be described below, these tests are distorted by the sampling solutions of the prior art, but are now possible when the sampling solutions of the present invention are used, even for the most delicate microorganisms, such as Listeria sp., or microorganisms present in low quantities (CFU).Thus, another related aspect of the present invention relates to the use of the composition or kit described above for the detection of contamination of the genus Salmonella on surfaces, preferably of the genus Salmonella enterica and / or Salmonella bongori and / or for the detection of contamination of the genus Listeria on surfaces, preferably of the genus Listeria monocytogenes and / or for the detection of contamination of the genus Pseudomonas on surfaces, preferably of the genus Pseudomonas aeruginosa and / or for the detection of contamination of the genus Staphylococcus on surfaces, preferably of the genus Staphylococcus aureus. In other words, this aspect of the present invention relates to the use of a liquid sampling composition comprising one or more surfactants and one or more (e.g.at least 2) enzymes selected from the group consisting of a protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase, for the detection of bacteria selected from Salmonella (e.g. Salmonella enterica, Salmonella bongori), Pseudomonas (e.g. Pseudomonas aeruginosa) and / or Listeria (e.g. Listeria monocytogenes) potentially present on a surface. Preferably, following this use, the detection is carried out by immunology or by genetic test and / or after culturing the bacteria. Preferably, in this use, the sampling composition comprises surfactants present at a content of between 0.02% and 1% (or even 1.5%; preferably between 0.1% and 0.5%) by weight relative to the weight of said sampling composition.Preferably, in this use, the surfactant(s) is (are) anionic, advantageously chosen from the group consisting of alkyl ether sulfate, preferably a salt of laureth sulfate, alkyl sulfate, alkylbenzene sulfonate and soaps, preferably the surfactants are essentially one or more anionic surfactants. Preferably, in this use, the sampling composition further comprises at least 15% of a stabilizing agent chosen from a polyol, preferably glycerol, and propylene glycol, relative to the weight of said sampling composition, and / or at least one sequestering agent (di- or multi-valent cations). Preferably, in this use, the sampling composition comprises a protease being a serine protease. Other characteristics, details and advantages of the invention will emerge from the description given below, in a non-limiting manner and with reference to the examples. Examples.The first obstacles that the inventors had to overcome concerned the lack of representativeness of the biofilms generated in the laboratory, compared to the situation on site. This complicates in particular the choice of enzymatic activities (nature of enzymes, their abundance, presence of potentially deleterious compounds). Based on these preliminary tests, the inventors therefore decided that all compositions that would be formulated in the laboratory should be tested in real conditions: many promising formulations at the laboratory scale are unusable, in practice, and the inventors have not found published guidelines, even with the help of their expertise, that could have helped them identify functional compositions.However, once a complex formulation has been identified, such as those described below, some compounds of this formulation can be replaced by others, which is easily testable in the laboratory (efficacy, absence of toxicity for microorganisms), then on site, or directly on site. Thus, to a certain extent, one protease is substitutable by another, in particular when these are serine proteases (subtilisins). However, when commercial enzymatic compositions are used, the inventors have noticed that, depending on the suppliers, and even the brands, different additives may interfere with microbiology analyses, thus variations of the composition of the present invention are possible, thanks to the teaching of the present invention, but require following the rules of caution described in the present invention.Example 1 A meat processing company located in Belgium was selected for this study. All facilities were pre-cleaned and disinfected before sampling.The detachment protocol is applied with a limitation of mechanical actions that can introduce a reproducibility error; it includes the following steps: Quickly scan (screen) the areas to be sampled with the UV lamp and BDK detection kit (Realco); Preheat the sampling solution to 45°C; Place two templates on the surfaces to be sampled (in order to delimit a surface of 10cm by 10cm): physiological water vs. sampling composition (14 sprays to cover the area; 1.2 g / spray); Leave the sampling solution to act for 5 minutes; Place the sterile wipe on the two sampling areas and let it soak; Collect sterile samples using forceps and place them in a sterile transport bag; Store the samples in refrigerators at 4°C; Evaluate the two areas with the lamp and the BDK.A total of 6 sampling areas were included in this study: Zone 1 round table; Zone 2 lid for trash table; Zone 3 stainless steel bin; Zone 4 saw table; Zone 5 plastic surface; Zone 6 derinding machine. In these 6 areas, the sampling composition comprising enzymes (including a subtilisin, activity equivalent to 10 Anson Units / kg) and detergents (approximately 10% by weight) allows a much better recovery of microorganisms, both in classical microbiology and by ATP (2G) measurement or by culture. Conversely, there were no more microorganisms left on the surfaces where the sampling solution was placed, unlike the surface treated with physiological water. The difference is especially marked at the level of classical microbiology, compared to ATP 2G or by test involving a culture step (few differences noted).The inventors reproduced the example, to include metagenomic analyses, while varying the contact time of the sampling composition on the surface (30 seconds and 5 minutes). Again, the enzymatic composition allows for much better sampling (e.g., 100 times better in conventional microbiology and 14 times better in qPCR). The bacteria found show an increased proportion of Pseudomonas. Example 2 – addition of a preservation solution; Salmonella The inventors, suspecting deleterious effects associated with the sampling composition, sought to determine to what extent an adjuvant solution could counteract these, in the context of tests aimed at determining the presence of Salmonella and / or Listeria, either by PCR or qPCR, or by using commercial detection kits, for example those from BioMérieux.To do this, a so-called 'preservation' solution was tested (in g / l): Proteins (beef extract; 5); Peptone, 10; Polysorbate 80 (Tween 80; 5); Lecithin 0.7; NaCl, 5. In addition, various serine protease inhibitors were used, with the aim of neutralizing the subtilisin activity of the sampling solution. The first series of tests was carried out in the laboratory, where known quantities of bacteria (0.1 ml of 10. 7 CFU / ml of Salmonella) were incubated either in physiological solution (100 ml), or in the collection composition of Example 1 (6 ml + 94 ml physiological water), or in the composition of Example 1, before the preservation solution was added (6 ml collection composition, 90 ml water, 4 ml of the preservation solution). When 10 7 CFU / ml of Salmonella are analyzed (therefore 10 4CFU / ml in the final mixture), the different situations are compatible with the identification of the microorganism by means of PCR analyses or the Vidas® Salmonella test. However, detection by the Vidas SLM® test (broth culture, then specific measurement by antigenic test and fluorescence; this test provides information on the absence or presence of Salmonella) fails when the sample composition is present, whereas it works when the bacteria are diluted in physiological saline or when the preservation solution has been added. Furthermore, in this example, the inventors consider that the Salmonella concentrations are high, which does not necessarily, or even rarely, represent the situation in the field, where even 1 CFU must be detected.Example 3 – Addition of a preservation solution; Listeria In this case, the inventors noticed that the enzymatic solution caused problems with detection (counting and qPCR); these problems were reduced thanks to the preservation solution, however the commercial Vidas® kit (LMX) still did not work. Furthermore, when the amount of Listeria inoculated was reduced, the microorganism was generally no longer detected following mixing (Vidas LMX®, never; Vidas LMO2®, not every time) with the sampling composition, whereas detection remained robust when mixing was carried out only in physiological saline. In this (intentionally extreme) case, the preservation composition did not improve detection. As a precaution, the tests were reproduced and three Listeria strains were tested with always the same results; these were ATCC 19114, NCTC 11994 and ATCC 13932.By comparing all the results, the inventors conclude that the sampling composition slows the growth of microorganisms (Listeria), a problem that is partially resolved by the preservation composition. On the other hand, the sampling composition interferes with certain downstream microbiological tests. Since the nature and abundance of microorganisms on a surface to be tested is unknown in advance, the inventors therefore conclude that the compositions of Examples 1 to 3 are useful, but would benefit from further improvement. Example 4 - Identification of deleterious compounds The inventors searched the various commercial enzymes to see if certain compounds interfered with downstream microbiological tests. The inventors noticed that the presence of compounds such as phenoxyethanol, potassium sorbate or thiazolinone derivatives is quite widespread.Thus, the inventors selected enzymatic compositions which are devoid of such compounds, or as poor as possible, even though the inventors found significant concentrations of 2-phenoxyethanol in many commercial enzymatic compositions. Among the compounds investigated are Bronopol, triclosan, idocarb, chlorhexidine, DCOIT, BIT, OIT, CIT, MIT, ethylparaben, methylparaben, butythylparaben, phenylparaben, isopropylparaben, isobutylparaben, propylparaben, salisylic acid, 4-hydroxybenzoic acid, 4-methylbenzoic acid, benzoic acid, sorbic acid, dehydroacetic acid, 1-phenoxypropan-2-ol, 1,2-dibromo-2,4-dicyanobutane, 2-hydroxybiphenyl, 2-phenoxyethanol (most abundantly found, sometimes up to 0.4% by weight), bensyl alcohol and chlorophenesin.Comparative Example 1 Suspecting interference between the proteases in the sample composition and the enzymes or antibodies in the downstream tests, the inventors tested the effect of adding protease inhibitors to the preservation solution (see Example 3), while avoiding as much as possible commercial enzymatic compositions comprising compounds identified as deleterious (see Example 4). Different ways of applying the protease inhibitor were tested. Since the chosen protease was a subtilisin or trypsin, two serine protease inhibitors were tested: PMSF and AEBSF. PMSF provided good inhibition of the proteolytic activities of the serine proteases tested. On the other hand, AEBSF only inhibited trypsin.Measurement tests where Listeria (the 3 strains above were tested) is brought into contact with a composition comprising a subtilisin, then AEBSF in a concentration sufficient to cause complete inhibition of subtilisin have shown that different microbiological tests gave lower values, or even did not detect Listeria under the test conditions, unlike the control without the enzymes or when PMSF is added. Thus, the inventors chose to incorporate PMSF into the preservation solution to be tested, despite the disadvantages of this molecule. Unfortunately, even under these conditions, the Vidas® LMX test does not allow the presence of Listeria monocytogenes to be detected. Example 5 – Optimization of the sampling composition The inventors then decided to test the effectiveness of a highly diluted sampling composition avoiding as much as possible the compounds identified as deleterious.Furthermore, enzymatic activities are reduced, as is the detergent content. Detergents are chosen after laboratory tests for their compatibility with the life of microorganisms, including Listeria, which has been identified as more sensitive. First, with detergent concentrations of 10% by weight, even in the presence of low enzymatic activity, in the presence of minimal preservative contents (e.g., almost entirely non-detectable; phenoxyethanol less than 4 ppm), and in the absence of PMSF, Listeria detection tests, such as Vidas®, do not work well (massive underquantification), or not reproducibly. Even less specific tests do not work when a Listeria culture is subjected to these different treatments.Thus, all the parameters of the composition must be checked, with the problem that too strong a dilution of the enzymatic activities risks no longer allowing the detachment of the microorganisms, while too strong a dilution of the detergents risks no longer allowing a homogeneous application of the composition or a good contact with the biofilm. Thus, the inventors noticed that, in addition to the quantity of the components, their nature had to be reconsidered. A diluted formulation was tested comprising approximately 20% by weight of glycerol, approximately 0.03% by weight of an anionic detergent of the laureth sulfate type (sodium), 0.4% by weight of sequestrant (carboxymethylinulin); 0.4% by weight of sodium glycolate; 0.04% by weight of a subtilisin (activity 1 Anson Unit), as well as a lipase; an amylase, a cellulase and a mannanase.The inventors selected enzymes, here commercial enzymes, free of preservatives such as potassium sorbate, MIT, OIT, BIT and CIT. When possible, the inventors used commercial enzymes free of phenoxyethanol (cellulase and mannanase), or with a low content of this molecule (subtilisin; 4 g / kg of the enzyme formulation, lipase; 0.249 g / kg and amylase; 0.233 g / kg). Enzymes other than subtilisin were added in weight contents of between 0.1 and 0.5% (weight of the enzyme composition: total weight) and the amount of enzyme is typically 1 to 10% by weight relative to the commercial formulation. Following the use of this composition, the presence of Listeria is detected, and the Vidas® LMX test for Listeria is, finally, positive and reproducible.The inventors further noted that, under these diluted conditions, the addition of the protease inhibitor PMSF is not necessary, and even reduces the propagation of Listeria. The inventors consider that the concentration of enzymes other than proteases (here, a subtilisin) can be further reduced somewhat, e.g., the lowest concentration that allows detachment, as can those of the sequestering agent. In light of the present invention, this can be tested in the laboratory on the 3 Listeria strains or in other models, including, preferably, more complex ones. The inventors then tested a possible synergy with the preservation solution (see examples 2 and 3): since the results are already very good, the improvement associated with the addition of the preservation solution is marginal, but it was nevertheless interesting in the case of a test that includes propagation followed by PCR detection.It is understood that the present invention is in no way limited to the embodiments described above and that many modifications may be made thereto without departing from the scope of the appended claims.

Claims

CLAIMS 1. A sterile sampling composition for microorganisms potentially present on a surface comprising: - one or more surfactants, present at a concentration of at least 0.02% by weight and at most 1.5% by weight relative to the weight of said composition (sum of the weights of surfactants / weight of the composition), said surfactant(s) comprising an anionic surfactant; - at least two enzymes selected from the group consisting of a (serine) protease, a polysaccharidase, a laccase, a lipase, a cellulase, and a mannanase; - at least one stabilizing agent, preferably present at a concentration of at least 15% relative to the weight of said composition, said stabilizer being selected from the group of polyols; - at least one sequestering agent. 2.A sampling composition according to claim 1, comprising at least one anionic surfactant selected from the group consisting of alkyl ether sulfates, preferably a laureth sulfate salt, alkyl sulfate, alkylbenzene sulfonate, and soaps.

3. A sampling composition according to claim 1 or claim 2, wherein the sequestering agent is selected from glucono-delta-lactone, sodium gluconate, potassium gluconate, calcium gluconate, citric acid, phosphoric acid, tartaric acid, sodium acetate, sorbitol, carboxymethyl inulin, and mixtures thereof.

4. A sampling composition according to any one of the preceding claims 1 to 3, comprising at least 50% (by weight) water, preferably at least 60% (by weight) water.

5. A sampling composition according to any one of the preceding claims 1 to 4, wherein the stabilizing agent is glycerol.

6. A sampling composition according to any one of the preceding claims 1 to 5 comprising at least one protease, preferably a serine protease, advantageously in a concentration equivalent to 0.5 to 5 Anson Units of Alcalase 2.5 DX per kg of said composition, more preferably in a concentration equivalent to about 1 Anson Unit of Alcalase 2.5 DX per kg of said composition. 7.A sampling composition according to any one of the preceding claims 1 to 6, having been sterilized by a physical method, preferably selected from gamma irradiation, electron beam irradiation, UV-C irradiation, sterilizing filtration, and combinations thereof, preferably sterilizing filtration followed by gamma irradiation, electron beam irradiation, or UV-C irradiation.

8. A sampling composition according to any one of the preceding claims 1 to 7, being in the form of a sprayable liquid. 9.Sampling composition according to any one of the preceding claims 1 to 8 comprising less than 100 ppm of 2-phenoxyethanol (parts per million: weight of 2-phenoxyethanol: weight of the composition), preferably less than 50 ppm of 2-phenoxyethanol, preferably less than 20 ppm of 2-phenoxyethanol, preferably less than 10 ppm of 2-phenoxyethanol, preferably less than 5 ppm of 2-phenoxyethanol, less than 4 ppm of 2-phenoxyethanol, or even less than 3 ppm of 2-phenoxyethanol.

10. Sampling composition according to any one of the preceding claims 1 to 9 comprising from 0.02 to. 0.5% of surfactant(s), preferably said surfactant(s) consisting essentially of one or more anionic surfactant(s).

11. Sampling kit for microorganisms potentially present on surfaces, in a living or viable state, for subsequent detection and laboratory analysis, comprising: - a sampling composition of surface microorganisms, preferably an industrial surface such as a food surface, an industrial surface such as, for example, the pharmaceutical industry, the healthcare environment, or the petrochemical industry, according to any one of claims 1 to 10, - at least one device for the controlled application of said sampling composition to said surface.

12. Sampling kit according to claim 11 further comprising at least one sterile container, for example, a sterile bag, and / or a sterile means for collecting microorganisms, preferably a tissue, a wipe, or a sponge. 13.A sampling kit according to claim 11 or 12, wherein the sampling composition comprises a serine protease and said kit further comprising a preservation solution for a sample taken, said preservation solution comprising a protein hydrolysate (of peptone) and / or a preservative inhibitor selected from polyoxyethylene sorbitan and / or a phosphoglyceride derivative.

14. A sampling kit according to any one of the preceding claims 11 to 13, further comprising means for the specific detection and / or specific quantification by immunology or genetics of bacteria, preferably selected from Salmonella (e.g., Salmonella enterica, Salmonella bongori), Pseudomonas (e.g., Pseudomonas aeruginosa), and / or Listeria (e.g., Listeria monocytogenes).

15. A method for sampling microorganisms on surfaces, in a living or viable state, for subsequent detection and laboratory analysis, comprising the following steps: a) at least one controlled application, preferably a spray of a controlled volume, of the microorganism sampling composition according to any one of claims 1 to 10, onto a surface to be sampled; b) allowing said composition to act on said surface to be sampled for a controlled period; c) bringing at least one sterile collection means for said microorganisms into contact with said surface to be sampled; d) allowing said at least one sterile collection means to be soaked in a controlled manner by said composition; e) aseptically collecting said at least one soaked microorganism collection means and depositing it into at least one sterile container, for example, a sterile bag containing a sample preservation solution comprising a protein hydrolysate.a protease inhibitor and / or a preservative inhibitor being polyoxyethylene sorbitan and / or a phosphoglyceride derivative, f) perform at least one microorganism detection test, said at least one detection test from among a test involving a microorganism culture step, a biochemical test, a test comprising an immunological recognition step, and a molecular biology test.

16. Use of the composition according to any one of claims 1 to 10 or of the kit according to any one of claims 11 to 14 for the detection of Salmonella contamination on surfaces, preferably of the species Salmonella enterica and / or Salmonella bongori.

17. Use of the composition according to any one of claims 1 to 10 or of the kit according to any one of claims 11 to 14 for the detection of Listeria contamination on surfaces, preferably Listeria monocytogenes.

18. Use of the composition according to any one of claims 1 to 10 or of the kit according to any one of claims 11 to 14 for the detection of Pseudomonas contamination on surfaces, preferably Pseudomonas aeruginosa.

19. Use of the composition according to any one of claims 1 to 10 or of the kit according to any one of claims 11 to 14 for the detection of Staphylococcus contamination on surfaces, preferably Staphylococcus aureus. 20.Use of a liquid sampling composition comprising one or more surfactants and one or more enzymes selected from the group consisting of a protease, a polysaccharidase, a laccase, a lipase, a cellulase, and a mannanase, for the detection of bacteria selected from Salmonella (e.g., Salmonella enterica, Salmonella bongori), Pseudomonas (e.g., Pseudomonas aeruginosa), and / or Listeria (e.g., Listeria monocytogenes) potentially present on a surface.

21. Use according to claim 20, wherein detection is performed by immunology or by genetic testing and / or after bacterial culture.

22. Use according to claim 20 or 21, wherein the sampling composition comprises surfactants present at a concentration of between 0.02% and 1% by weight relative to the weight of said sampling composition.

23. The use according to claim 22, wherein the surfactants comprise an anionic surfactant, preferably said anionic surfactant being selected from the group consisting of alkyl ether sulfate, preferably a laureth sulfate salt, alkyl sulfate, alkylbenzene sulfonate, and soaps, preferably the surfactants being essentially one or more anionic surfactants.

24. The use according to any one of the preceding claims 20 to 23, wherein the sampling composition further comprises at least 15% of a stabilizing agent selected from a polyol, preferably glycerol, and propylene glycol, by weight of said sampling composition.

25. The use according to any one of the preceding claims 20 to 24, wherein the sampling composition comprises a protease being a serine protease.