Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
  • A01N63/50—Isolated enzymes; Isolated proteins
• • 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
  • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
• • 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
  • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
  • C11D3/38618—Protease or amylase in liquid compositions only
• • 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
  • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
  • C11D3/38636—Preparations containing enzymes, e.g. protease or amylase containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase
• • 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
  • C11D3/48—Medical, disinfecting agents, disinfecting, antibacterial, germicidal or antimicrobial compositions
• • 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
  • C11D2111/00—Cleaning compositions characterised by the objects to be cleaned; Cleaning compositions characterised by non-standard cleaning or washing processes
  • C11D2111/40—Specific cleaning or washing processes
  • C11D2111/44—Multi-step processes
• • 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
  • C11D2111/00—Cleaning compositions characterised by the objects to be cleaned; Cleaning compositions characterised by non-standard cleaning or washing processes
  • C11D2111/40—Specific cleaning or washing processes
  • C11D2111/46—Specific cleaning or washing processes applying energy, e.g. irradiation

Definitions

• the present invention relates to a method for eliminating biofilms for cleaning medical instruments, in particular for combating nosocomial diseases.
• a biofilm is a viscous film that develops on all surfaces, following the adhesion of microorganisms on these surfaces and the secretion by them of polymers that cover them and facilitate their adhesion.
• Biofilms thus constitute a protective layer around microorganisms and represent a recurrent source of contamination of the surrounding environment which poses major problems in terms of health, for example in hospital environments.
• Biofilms are found in many environments, for example in water circuits, cooling towers, on any immersed material (boat hull, ...) but also on the teeth (plate) or in the intestines and especially on medical instruments used in hospitals.
• this matrix is very resistant and can constitute a barrier protecting the bacteria agents that act against microorganisms.
• Conventional treatments based on soda and / or with different biocides do not act sufficiently efficiently because they do not penetrate the biofilm throughout its thickness or are inhibited by certain molecules making up this matrix. The treatment is then only partially effective on the upper surface of the biofilm.
• WO2012 / 048757 discloses a composition and a method for eliminating biofilms for cleaning floors and surfaces in the agro-food industries. More particularly, the composition disclosed in this document is intended for the treatment and cleaning of installations in the field of the food industry where bacteria specific to this sector of activity secrete protective biofilms responsible for a recurrent source of contamination of food and / or facilities of the agri-food industries.
• biofilms In the field of the food industry, but even more so in hospital, the problem of the presence of biofilms is twofold. First, they represent a source of permanent contamination and very difficult to eliminate by conventional means, even the most aggressive ones. Indeed, conventional disinfectants are ineffective because they fail to reach the microorganisms that are protected by the biofilm. Their incomplete elimination causes and accelerates patient-to-patient contamination transfers as soon as they are virtually invisible. However, as a result of shocks or friction, film of biofilms can come off and release bacteria that were housed there. Indeed, these hidden bacteria are for example released following an impact or friction of a surgical tool on a carriage or following the rubbing of a bed against a door, which allows a free transfer of bacteria to a patient often in fragile condition.
• a biofilm is mixed. That is to say that, developed by certain strains, a biofilm can shelter other strains, which live and then develop in colonies. These colonies promote communication between bacteria and, among other things, gene exchange. The propagation of resistance genes carried by certain bacteria is thus facilitated within the biofilms which are then even more difficult to eliminate.
• hospitals provide a particularly favorable environment for gene exchanges since many bacteria coexist there, for example in rooms, in the operating rooms but also on medical instruments (orthopedic graters, reamers, cannulas, endoscopes, ). In biofilms, the problem is further accentuated because these strains are not even reached by the disinfectants protected by the biofilm matrix.
• the object of the invention is to overcome these disadvantages by providing a method as indicated at the beginning, characterized in that it comprises a surface treatment step for a predetermined period of time with a composition comprising at least one detergent component and at least one enzymatic component and a step of rinsing and / or drying said surface.
• said detergent component comprises a wetting agent and a dispersing agent.
• said detergent component further comprises a sequestering agent.
• said at least one enzymatic component contains for example at least one protease, at least one laccase and at least one polysaccharidase.
• said wetting agent and said dispersing agent form a premix and are thus mixed together.
• such a treatment with such a composition according to the invention allows the enzymes (proteases, laccases, polysaccharidases) to effectively and polyvalently degrade the organic polymers of different natures constituting the matrix of biofilms formed by a multitude of different microorganisms directly or indirectly responsible for nosocomial diseases.
• the enzymes protes, laccases, polysaccharidases
• the material of the biofilm is weakened and swollen, allowing it to be removed from the treated surface.
• the process according to the invention is not specific to a particular microorganism and therefore to a particular type of biofilm but that it is suitable for many bacterial strains, the enzymes acting on matrix polymers of biofilms formed by any microorganism.
• the action detergent of the composition according to the invention also makes it possible to ensure the effectiveness of the composition according to the invention.
• a detergent base which is compatible and which can act synergistically with the enzymatic activity of the enzyme component.
• a detergent base that significantly improves the speed and efficiency of the elimination of biofilm.
• the present invention combines a wetting agent and a dispersing agent and optionally a sequestering agent.
• the joint actions of these agents of the detergent component of the composition according to the invention makes it possible to eliminate the superficial part of the biofilm, to wet and to swell the organic structures of the biofilm, thus in this way promoting the accessibility of the enzymatic component which weakens and in turn degrades the matrix of the biofilm.
• said surface treatment step for combating nosocomial diseases comprises a step of eliminating biofilms making it possible to reduce the microorganisms responsible for nosocomial diseases.
• a log reduction (or reduction) of at least 1 log of the microorganisms protected by biofilms is significant for the purpose of controlling nosocomial diseases, which corresponds to a percentage of at least 90% reduction of microorganisms protected by biofilms.
• This log reduction is calculated by converting the microbial load to log 10, subtracting the test sample result from the control (control) sample, i.e. subtracting the microbial load present in a given environment (for example on the surface of a medical tool or on the surface of a soil) before a surface treatment step according to the invention of the microbial load present in the same environment following a surface treatment step according to the invention. invention.
• said surface treatment step is carried out by dipping for a predetermined period of time of between 20 minutes and 24 hours in a dipping solution comprising said composition and a previously formed aqueous dilution phase.
• a soaking treatment is particularly suitable for cleaning surfaces or medical tools (orthopedic graters, reamers, cannulas, endoscopes ...) as long as these surfaces or tools can be immersed in a container (tank, basin, etc.). ) filled with said dipping solution.
• the medical tools may stay longer or shorter, for example minutes, hours or days in the dipping solution without impeding the implementation of other medical interventions.
• said dipping solution can be injected into the endoscope tubes for cleaning.
• said step of surface treatment by dipping is associated with an abrasion step mechanically said surface with said dipping solution, for example by mechanized or manual brushing or by sonication.
• An additional mechanical abrasion step allows the soaking solution comprising said composition in the aqueous phase to act on the different layers of the biofilms but also to participate mechanically in the destructuring of the polymer matrix and thus remove films from the surface of the biofilms so that the enzymes and other components of said composition achieve even better the different layers of the biofilms, which ensures an optimal treatment of the surfaces in order to effectively remove the biofilms.
• the pH of said dipping solution is adjusted to a pH value of between 7 and 8.
• the method further comprises a further subsequent step of treating said surface with a biocide.
• An additional disinfecting biocide treatment following the action of the enzymatic solution during the step of surface treatment by dipping, ensures the destruction of the bacteria made free at the end of treatment of said surface.
• this treatment step using a biocidal agent may be followed by a sterilization step, for example by autoclaving.
• the present invention also relates to a new use of a composition comprising at least one detergent component and at least one enzymatic component, for combating nosocomial diseases by eliminating biofilms.
• Removal of the biofilms by soaking in the diluted composition makes it possible to fight effectively against nosocomial diseases since the enzymatic components of the aqueous dipping solution make it possible to Destructure and eliminate biofilms from treated surfaces so that biofilms, largely responsible for nosocomial diseases, are eliminated.
• This action of the enzymes is favored by the fact that it takes place in synergy with the action of the detergent component of the composition according to the invention, which makes it possible to eliminate the superficial part of the biofilm, to wet and to swell the organic structures. biofiim and thus promote the accessibility of the enzymatic component which in turn weakens and degrades the biofilm matrix.
• said detergent component of the composition used comprises a wetting agent and a dispersing agent.
• said detergent component of the composition used further comprises a sequestering agent.
• said enzymatic component of the composition used contains for example at least one protease, at least one laccase and at least one polysaccharidase.
• the present invention relates to a use of a composition
• a composition comprising at least one detergent component comprising a wetting agent and a dispersing agent and optionally a sequestering agent, and at least one enzymatic component containing at least one protease, at least one laccase and at least one polysaccharidase for the removal of sources of contamination including Staphylococcus aureus and / or Escherichia coli and / or Pseudomonas aeruginosa. These bacteria grow particularly well in hospitals and are responsible for many infections.
• staphylococcus aureus found in 15 to 30% of healthy individuals, is very resistant and is transmitted very quickly from one patient to another not only by direct transmission via the caregiver but also by indirect transmission via objects (medical tools, ...) and even dust.
• Staphylococcus aureus is considered to be the main bacterium responsible for hospital infections and it is therefore necessary to control its responsible spread, in large part, of nosocomial diseases leading to the death of the patient.
• the invention also relates to a use of a composition comprising at least one detergent component and at least one enzymatic component, for the treatment of surfaces, medical tools or floors in order to fight nosocomial diseases by soaking.
• the detergent component of said used composition comprises a wetting agent and a dispersing agent.
• said detergent component of said composition used further comprising a sequestering agent.
• said enzymatic component of said composition used contains for example at least one protease, at least one laccase and at least one polysaccharidase.
• the detergent component comprising a wetting agent and a dispersing agent and optionally a sequestering agent acts first by removing a surface portion of the biofilm and by wetting and / or swelling the organic structures of the biofilm.
• the detergent component thus promotes the accessibility of the enzymatic component by destructuring the matrix of the biofilm.
• the enzymatic component then acts synergistically with the detergent component and weakens and in turn degrades the biofilm matrix.
• This combined action of the three types of enzyme and the detergent component perfectly compatible with a correct action of the enzymes, favors the accessibility of the composition of the deeper layers and allows a fast and optimal detachment of any type of biofilm while preserving the substrate.
• Such a composition according to the invention further reduces the input of external compounds into the installations during the cleaning step and thus simplifies the validation procedures of the cleaning steps.
• the detergent component allows enzymes to act quickly on all structures of biofilms, which, in a hospital environment where tools, rooms and operating rooms must be reused very quickly following an intervention, represents a definite advantage.
• the dispersant of the detergent component improves the separation of particles from a slurry to prevent agglutination, aggregation and / or decantation.
• Said dispersing agent may be a sotoluble or partially water-soluble polymer such as, for example, polyethylene glycol, cellulose derivatives or a polymer comprising at least one acrylic acid or acrylic ester unit.
• the dispersing agent is a polymer comprising at least one acrylic acid or acrylic ester unit of general formula - (CH 2 -CH-COOR) - in which R represents a hydrogen, alkyl or substituted alkyl, aryl or substituted aryl group.
• the dispersing agent is a polymer having a mean molecular weight Mw of approximately between 500 and 0000.
• the dispersing agent is a polymer of acrylic acid.
• the dispersing agent may be a homopolymer of acrylic acid having a mean molecular weight of approximately between 2000 and 6000.
• the detergent component comprises a proportion of dispersing agent of between 1 and 10% by weight relative to the total weight of the detergent component.
• said dispersing agent of said detergent component is C 6 alkyl kglucoside.
• composition according to the invention for eliminating biofilms and combating diseases nosocomiales avoids any aggregation of bacterial particles during the cleaning of surfaces, which ensures optimal removal of biofilm particles detached from a support under the action of enzymes. Indeed, rather than being aggregated, these particles remain separated in a suspension, do not redeposit and do not re-adhere to the cleaned support.
• the wetting agent of the detergent component is an amphiphilic chemical substance, or a composition comprising said amphiphilic chemical substance, which modifies the surface tension between two surfaces.
• the wetting agent has the advantage of promoting the spreading of a liquid on a solid but also to accentuate the contact between two surfaces. More particularly, the wetting agent promotes contact between the detergent component and a surface and, therefore, between the enzymes and their substrate.
• the surfaces to be treated are often stainless steel or a material on which the application of a liquid gives rise to the formation of droplets. This characteristic of the surfaces to be treated makes it difficult to homogeneously apply a composition in liquid form to combat the presence of biofilms.
• the wetting agent is a particularly advantageous component of said composition according to the invention since it allows, even on surfaces of the stainless steel type, a homogeneous spread of the composition and thus its perfect distribution on the surfaces to be decontaminated. eg on surgical tools and soils.
• the wetting agent may be anionic, cationic, nonionic or zwitterionic.
• the wetting agent may be anionic or nonionic wetting agent, that is to say that the hydrophilic part is negatively charged or has no net charge, or may be a composition comprising an anionic wetting agent.
• the wetting agent may be a sucrose ester or a composition comprising sodium alkyl sulphate and an alcohol.
• said wetting agent is non-foaming hot and is preferably selected from the group of C 6 to C 10 sodium alkyl sulphates, ethereal alcohol sulphates. -C10 and sulfonates alkylearyles Ce to Ci 0.
• said wetting agent of said detergent component is ethoxylated 2-ethylhexanol.
• the wetting agent is non-foaming hot allows use of the composition according to the invention for the treatment of medical tools with pipes, such as endoscopes. Indeed, the wetting agent being non-foaming, this prevents the formation of foam, without altering, on the contrary, the surfactant and / or emulsifying performance of the composition according to the invention. It is understood that the provision of an effective detergent solution without foam generation limits rinsing steps, which is particularly desirable, especially for medical tools with pipes that can be reused quickly for a new intervention.
• the detergent component comprises a wetting agent content of from 1 to 15% by weight based on the total weight of the detergent component.
• the sequestering agent is a chemical substance having the ability to form complexes with mineral ions which it fixes in a form preventing their precipitation by the usual reactions.
• the sequestering agent may be ethylene-diamine tetraacetic acid, glucono-delta-lactone, sodium gluconate, potassium gluconate, calcium gluconate, citric acid, phosphoric acid, tartaric acid, sodium acetate, sorbitol, a compound having a phosphorus atom.
• the agent 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 bearing at least, in its structure, a phosphine functional group , phosphine oxide, phophinite, phosphonite, phosphite, phosphonate, phosphinate or phosphate, alone or in combination, or a salt thereof.
• 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 bearing at least, in its structure, a phosphine functional group , phosphine oxide, phophinite, phosphonite, phosphite, phosphonate, phosphinate or phosphate, alone or in combination, or a salt thereof.
• the sequestering agent is a chemical substance compatible with the substances that can be used in hospitals, for example, the sequestering agent is preferably an agent that is not toxic to human health.
• the sequestering agent may be a phosphonate or a salt thereof, an amine or an amine oxide comprising at least, in its structure, a functional group phosphine, phosphine oxide, phosphinite, phosphonite, phosphite, phosphonate, phosphinate or phosphate, alone or in combination, or a salt thereof.
• the amine or the amine oxide may comprise one, two or three substituents of general formula CR 4 R 5 W in which R 4 and R 5 independently represent one of the other is hydrogen, alkyl, substituted alkyl, substituted or unsubstituted alkylamino, substituted or unsubstituted aminoalkyl, aryl or substituted aryl, and W represents a phosphonate, phosphinate or phosphate group.
• the sequestering agent may be in the form of a salt of sodium, calcium, lithium, magnesium or potassium; preferentially, the sequestering agent may be in the form of a sodium, calcium or potassium salt.
• the sequestering agent is an agent that can be safely used in medical procedures, i.e. the sequestering agent does not present a health hazard, alone or in combination with other components used in hospital.
• said sequestering agent of said detergent component is a potassium salt based on modified phosphonic acid (N-oxide ATMP).
• said at least one detergent component comprises a proportion of sequestering agent of between 1 and 10% by weight relative to the total weight of the detergent component, which represents an optimum between efficiency, stability and cost.
• said at least one enzymatic component comprises a proportion of protease (s) of between 10 and 50%, a proportion of laccase (s) of between 5 and 35% and a proportion of polysaccharidase (s) of between 5 and 20%. % by weight relative to the total weight of the enzymatic component, the 100% of the enzymatic component being optionally achieved using a conventional excipient or solvent, for example an alcohol.
• the enzymatic component according to the invention has the advantage of being polyvalent, that is to say that it can act simultaneously on various bacterial strains, which is particularly advantageous in a hospital environment where many different bacterial strains develop. simultaneously.
• the enzymatic component may contain between 1 and 10 proteases, preferably between 1 and 5 proteases, more preferably it may contain 2, 3, 4 or 5 proteases.
• Nonlimiting examples of protease enzymes belonging to class EC 3.4 and capable of being used in the invention are amino peptidases (EC 3.4.1 1), dipeptidases (EC 3.4.13), dipeptidyl peptidases and tripeptidyl peptidases (EC 3.4.14), peptidyi-dipeptidases (EC 3.4.15), serine carboxypeptidases (EC 3.4.16), metallo carboxypeptidases (EC 3.4.17), cysteine carboxypeptidases (EC 3.4.18), omega peptidases (EC 3.4.19), serine endopeptidases (EC 3.4.21), cysteine endopeptidases (EC 3.4.22), aspartic endopeptidases (EC 3.4.23), metalloendopeptidases (EC 3.4.24), threonine endopeptidases (EC 3.4.25), and endopeptidases in EC 3.4.99.
• amino peptidases EC 3.4.1 1
• dipeptidases EC
• the proteases belong to the class EC 3.4.21.
• Proteases are available commercially and in various forms including powders, granules, suspensions, liquid solutions.
• laccases used in the invention belong to the class EC 1.10.3.2.
• Laccases are copper-containing enzymes whose function is to oxidize a substrate in the presence of oxygen. More specifically, laccases are oxidoreductases that work with molecular oxygen as an electron acceptor.
• the at least one polysaccharidase used in the invention is an enzyme whose function is to break bonds within polysaccharides.
• the at least one polysaccharidase may be alpha-amylase, chiluiase, hemi-cellulase, glucosidase, beta-glucanase or pectinase.
• the at least one polysaccharidase may be an alpha-amylase belonging to the class EC 3.2.1.1, whose function is to break (1-4) -alpha-glycoside bonds in polysaccharides containing three units or plus alpha- (1-4) -D-glucoside.
• the enzymatic component may comprise a proportion of laccase (s) of approximately 30%, a proportion of protease (s) of approximately 30%, a proportion of alpha-amylase (s) of approximately 10% by weight relative to total weight of the enzymatic component, the 100% of the enzymatic component possibly being reached with the aid of a conventional excipient or solvent.
• laccase s
• protease s
• alpha-amylase s
• the proportion of laccase (s) may be approximately 30%, the total proportion of proteases approximately 30%, the proportion of alpha-amylase ( s) approximately 10% by weight relative to the total weight of the enzymatic component, the 100% of the enzymatic component possibly being reached with the aid of a conventional excipient or solvent.
• the ratio of each protease can range from 1: 2 to 2: 1, preferably the ratio of each protease can be 1: 1.
• the enzymes present in the enzymatic component have an additional action on the biofilm.
• laccase is highly effective on soils not attacked by alpha-amylase or proteases.
• the enzymatic component by the simultaneous presence of at least three types of enzymes, is versatile and can act at the same time on various types of biofilms produced by various bacterial strains, which is essential in the environment. hospital.
• the enzymatic component may be a solution or in solid form.
• the enzymatic component is a solution whose pH can be between approximately 8 and 10.
• the enzymatic component is an aqueous solution whose pH can be approximately between 8.5 and 9.5; more preferably the pH can be approximately 9.0, and this to preserve the integrity of the enzymes as much as possible.
• the enzymatic component may be in solid form such as, for example, in the form of a lyophilizate, powders, granules or in any other form permitting the solubilization of said component in a solvent, then it will subsequently be dissolved in said solvent.
• the solvent may be water or an aqueous, acidic, basic, alcoholic, buffered or neutral solution.
• the enzymatic component solubilized can then in this case be further diluted in an aqueous solution optionally containing one or more compounds such as for example detergents to form the cleaning solution.
• the detergent component may be in solid form to be dissolved in a solvent and / or in an aqueous phase or in liquid form.
• the enzymatic component When it is in solid form, it can either be put directly in solution in the solution formed by the enzymatic component possibly already diluted in the aqueous phase, or be dissolved in a solvent, prior to its dilution in the solution formed by the enzymatic component and the aqueous phase, or in the aqueous phase directly, before the dilution of the enzymatic component.
• 100% of the detergent component is optionally reached generally with water and, prior to application on the biofilm, it will be diluted in an aqueous phase, possibly containing the enzyme component.
• the pH of said solution is adjusted to a pH value between 6.5 and 7.5, more particularly around 7.
• said composition may be in solid form and then dissolved before use in a solvent in order to obtain, when it is diluted in an aqueous phase before application to a biofilm, a solution whose pH is approximately between 7 and 8.
• the composition is a solution intended, when it is diluted in an aqueous phase before application to the biofilm, to form a solution having a pH approximately between 6.5 and 7.5, more particularly of about 7.
• the pH of the solution of the composition is particularly suitable for the action of the enzymatic component, in particular laccase.
• said composition may be in solid form and then dissolved before use in a solvent to obtain a solution which will then be diluted in an aqueous phase to obtain a cleaning solution having a pH of approximately 6.5 to 7, 5, preferably about 7.
• the entire matrix of biofilms is removed, the surface treatment according to the invention leaving only "bare" bacteria, that is to say bacteria without any protection. and which can thus be easily removed in full during a subsequent use of a biocide, as explained hereinafter after rinsing the composition.
• the composition according to the invention comprises a detergent component that promotes the action of enzymes by allowing them to quickly reach the lower layers of biofilms and not only their upper layer. Enzymes can therefore act very rapidly on the entire biofilm structure and thus expose all the bacteria not only to the surface of the biofilm but also to the lower layers that make it up.
• FIGS. 1a to 1f illustrate the colorations obtained (the zones surrounded are zones having a blue coloration) and the use of a detection kit for the presence of biofilm to evaluate the effectiveness of cleaning techniques on rasps. with narrow pores after pre-cleaning (1a), after cleaning in an Aniosyme® solution (1b) and after cleaning in a dipping solution according to the invention (1c) but also for wide-pore graters after pre-cleaning (1a) -cleaning (1d), after cleaning in an Aniosyme® solution (1e) and after cleaning in a dipping solution according to the invention (1f).
• 2a to 2f illustrate the colorations obtained (the zones surrounded are zones having a blue coloration) when using a detection kit according to patent EP 2537601 for the presence of biofilm to evaluate the effectiveness of the techniques of cleaning on rasps after pre-cleaning (2a), after cleaning in a dipping solution according to the invention (2b), after cleaning in a dipping solution according to the invention and brushing (2c), after cleaning in a solution of Aniosyme® (2d), after cleaning in Aniosyme® solution and brushing (2nd) and after cleaning in an Aniosyme® solution and brushing then cleaning in a soaking solution according to the invention and brushing (2f).
• Example 1 Cleaning medical instruments by soaking in a dipping solution according to the invention - ATP-metric analyzes
• Tests have been carried out to determine whether the method according to the invention makes it possible to effectively remove biofilms present on medical instruments by soaking in an aqueous dipping solution according to the invention.
• This aqueous soaking solution was prepared by diluting a first product comprising a detergent component comprising a sequestering agent, a wetting agent and a dispersing agent and a second product comprising an enzymatic component comprising at least one protease, at least one laccase and at least one polysaccharidase in water of distribution at 45 ° C so that the aqueous soaking solution comprises a concentration of 0.25% (v / v) of said first product and a concentration of 0.1 % (v / v) of said second product.
• ATP-metric analyzes to measure the presence of adenosine triphosphate, were performed to determine:
• the amount of ATP molecules on the surface of the medical instruments was determined by swabbing the surface of the instruments before applying the 3M TM ATM Testing Protocol (3M Clean-Trace).
• This analysis technique makes it possible to measure a quantity of light produced (URL) which is directly proportional to the amount of biological energy present in the analyzed solution, that is to say to the quantity (concentration) of microorganisms in the solution analyzed.
• the measurement of the quantity of light emitted thus makes it possible to highlight and quantify the presence of ATP molecules and thus to determine the amount (concentration) of microorganisms in the analyzed solution (since ⁇ is a molecule essential to life microbial).
• ATP-metric analyzes have determined a significant amount of biofilms (ATP molecules) for the reamer before cleaning by dipping (1576 URLs) while low levels of fouling (contamination by the presence of biofilms) have been detected. recorded for other medical instruments that have therefore not been subjected to a soaking wash in the aqueous soaking solution according to the invention.
• the results obtained with the reamer make it possible to note that the level of ATP measured directly after immersion of the reamer in the soaking solution is very low (14 URLs) but that this level increases significantly after 30 minutes of soaking in the soaking solution. This clearly indicates that soaking in the soaking solution according to the invention makes it possible to unhook the biofilms from the surface of the medical instruments, in this case the reamer in the present case, which makes it possible to access the microorganisms.
• Comparative tests were carried out to compare, in terms of elimination of biofilms, a cleaning carried out by soaking for 30 minutes in an aqueous dipping solution according to the invention (as described in Example 1) with a cleaning carried out by autoclaving at 134 ° C for 90 minutes (currently used in hospital).
• the detection kit consists of two products, a staining reagent and a cleaning reagent.
• the blue dye present in the staining reagent specifically adheres to EPS (Extracellular Polymeric Substances) forming the protective matrix of the biofilm.
• EPS Extracellular Polymeric Substances
• the surface to be analyzed is sprayed with the staining solution. After 5 minutes of action on the surface, the excess of the solution is absorbed. Then the surface is rinsed with the cleaning solution which requires a five minute exposure time. The surface is finally rinsed with water and analyzed.
• the presence or absence of biofilm is demonstrated by a more or less dark blue color depending on the amount of btofilm existing on the surface. An intense blue color indicates a significant presence of biofilm.
• Comparative tests were carried out to compare, in terms of elimination of biofilms, a cleaning performed by soaking for 30 minutes in an aqueous solution according to the invention (as described and prepared in Example 1), a cleaning performed. by soaking for 5 minutes in an aqueous solution of Aniosyme® (a solution claiming to eliminate biofilms) 0.5% prepared according to the manufacturer's instructions and cleaning carried out by autoclaving at 134 ° C for 90 minutes. Prior to these three types of cleaning, the medical instruments were first pre-cleaned in a cleaning-disinfector so that they were ready for use according to hospital procedures and standards of application.
• ATP-metric analyzes according to the analysis protocol of the ATP Testing method proposed by 3M TM (3M Clean-Trace), were carried out to measure:
• a very high ATP value (2487.5 URL) is observed for the suction cannula for laparoscopy prior to cleanings. This indicates a large amount of residual biofilms after pre-cleaning in a cleaner-disinfector and therefore the instruments supposedly ready for use still have biofilms protecting microorganisms potentially responsible for nosocomial diseases.
• the ATP level goes down to a very low level (9 URLs), indicating a high level of cleanliness, corresponding to a reduction logarithmic of 1, 6 log microorganisms protected by biofilms and responsible for diseases nosocomiales, that is to say a percentage reduction of more than 90% of microorganisms protected by biofilms.
• Similar results are obtained for the large ENT cannula for which a log reduction of 2.44 Log is noted, which corresponds to a reduction percentage greater than 90% of microorganisms protected by biofilms.
• the other instruments analyzed have low to medium ATP values before cleaning and have therefore not been subjected to cleaning with the dipping solution according to the invention.
• EXAMPLE 4 Comparative Tests for Cleaning Medical Instruments by Soaking in a Soaking Solution According to the Invention and by Soaking in an Aniosyme® Water Solution - Analyzes Using a Detection Kit
• Comparative tests were carried out to compare, in terms of elimination of biofilms, a cleaning performed by soaking for 30 minutes in an aqueous solution according to the invention (as described and prepared in Example 1), a cleaning performed. by soaking for 15 minutes in an aqueous solution of Aniosyme® (a solution claiming to eliminate biofilms) at 0.5% prepared according to the manufacturer's instructions.
• Example 2 To measure the effectiveness of each of these cleanings, a detection kit as described in Example 2 was used on the surfaces of the medical instruments:
• biofilm The presence or absence of biofilm is demonstrated by a more or less dark blue color depending on the amount of biofilm existing on the surface. An intense blue color indicates a significant presence of biofilm.
• Comparative tests were carried out to compare, in terms of elimination of biofilms, a cleaning performed by soaking for 30 minutes in an aqueous dipping solution according to the invention (as described in Example 1) and associated with a step manual scrubbing to a cleaning performed by soaking for 15 minutes in an aqueous solution of Aniosyme® (a solution claiming to eliminate biofilms) to 0.5% prepared according to the manufacturer's instructions and associated with a manual brushing step.
• Aniosyme® a solution claiming to eliminate biofilms
• Example 2 To measure the effectiveness of each of these cleanings, a detection kit as described in Example 2 was used on the surfaces of the rasps: - before cleaning,
• biofilm The presence or absence of biofilm is demonstrated by a more or less dark blue color depending on the amount of btofilm existing on the surface. An intense blue color indicates a significant presence of biofilm.
• pre-cleaning supposed to provide ready-to-use graters leaves biofilms (medium to intense coloration depending on the size of the pores of the graters).
• Example 6 Cleaning endoscopes with a dipping solution according to the invention
• a first step consisted in performing a blanking of the endoscopes in a lava endoscope according to a protocol and a known procedure with commercial solutions Soluscope E (enzymatic solution) at 0.5% and Soluscope D (disinfectant solution based on glutaraldehyde) according to the manufacturer's instructions.
• a third step concerned the cleaning of endoscopes in the Save endoscope with a solution according to the invention (as described in Example 1) and with a commercial disinfectant solution (Soluscope D) following the two previous steps.
• This cleaning in the endoscope washer consists of a cycle of 35 minutes comprising the following sequential steps: leak test - pre-cleaning - cleaning - rinsing - disinfection - rinsing - drying.
• the counts of bacteria developing on petri dishes have made it possible to demonstrate a strong presence of microorganisms at the neck of the endoscope lava (red colonies) and at the level of the evacuation grid (colony yellow-gold) and filter (red colony).
• the ATP-metric analyzes revealed a low presence of microorganisms on these same areas of the endoscope lava (URL values less than 500).
• Bacterial colonies were counted on the non-selective culture medium (28 CFU / 150 ⁇ l) and then subcultured on a selective medium for growth of staphylococci, which established the presence of Staphylococcus aureus in the lava rinsing water. endoscope after cleaning with the dipping solution according to the invention. The Bart Test measurements confirmed this presence of bacteria in the rinsing water of the endoscope lava.
• Bacterial colonies were counted in the rinse water after cleaning the endoscopes with the solution according to the invention (red colonies, 32 C FU / 150 I). The bacteria forming these colonies have been identified as Staphylococcus aureus. Bart Test measurements also indicated the presence of bacteria in the rinse water.

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